Dissertations / Theses: 'Amyloliquefaciens'

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Makarewicz, Oliwia. "Regulation der Phytaseexpression in Bacillus amyloliquefaciens." Doctoral thesis, [S.l.] : [s.n.], 2006. http://deposit.ddb.de/cgi-bin/dokserv?idn=983031339.

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Lima, Frederico Alves. "Produção de biossurfactantes por Bacillus amyloliquefaciens IT45." Universidade Federal de Uberlândia, 2017. http://dx.doi.org/10.14393/ufu.di.2017.62.

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Biossurfactantes são moléculas de origem microbiana que possuem importante ação na redução tensão superficial. Dentre os biossurfactantes mais efetivos estão os lipopeptídeos produzidos por bactérias do gênero Bacillus, especialmente a Surfactina. Estes biocompostos apresentam uma série de vantagens que potencializam suas aplicações, tais como: estabilidade frente a condições extremas (pH, temperatura), diversidade de estruturas químicas, excelentes propriedades superficiais e ecológicas, ação antibiótica frente a microrganismos patógenos, dentre outras. Diante deste contexto, neste trabalho foi avaliada a produção de biossurfactantes totais e a Surfactina por Bacillus amyloliquefaciens IT45, quando variada a concentração dos reagentes presentes no meio de cultivo. As fermentações submersas foram realizadas em mesa agitadora industrial e em biorreator piloto de capacidade 40 litros. Para aperfeiçoar a concentração dos reagentes presentes no meio de cultura, um Planejamento Composto Central foi desenvolvido com propósito de avaliar a influência de três variáveis (xarope de glicose, extrato de levedura e cloreto de cálcio) na tensão superficial, na produção de biossurfactantes totais e no açúcar residual. Depois das análises estatísticas, quando as variáveis estavam nas concentrações (g.L-1) de 20 para xarope de glicose, 15 para extrato de levedura e 4 para cloreto de cálcio, a tensão superficial (mN/m) foi reduzida de valores acima de 50 para 30, o açúcar residual foi mínimo e igual a 31% e a produção de biossurfactantes totais foi máxima e igual a 5,5 g.L-1, depois de um período de cultivo de 48 horas. A caracterização do biossurfactante sugeriu a presença da Surfactina e este composto foi quantificado no tempo de retenção de 13,5 minutos. Com intuito de saber a real produção de Surfactina e crescimento biomassa celular, foram feitas fermentações em biorreator piloto de 40 litros e os resultados mostraram bastantes favoráveis. O tratamento com maior destaque foi referente à receita sugerida pelo Planejamento Composto Central em que o xarope de glicose, extrato de levedura e cloreto de cálcio estavam nas concentrações (g.L-1) de 20, 15 e 4, respectivamente. Neste cultivo o crescimento celular de 6,0 x 109 CFU.mL-1, produção de Surfactina de 0,63 g.L-1 e açúcar residual de 28%. Também foi realizado teste de atividade antimicrobiana contra 5 fungos patogênicos de diferentes gêneros. O caldo fermentado livre de células mostrou-se promissor, pois causou inibição em 4 fungos dos 5 estudos. Portanto, os resultados demonstram que o Bacillus amyloliquefaciens IT45 tem potencial para produção de biocompostos, uma vez que não necessita de altas concentrações de fonte de carbono e nitrogênio para seu desenvolvimento.
Biosurfactants are molecules of microbial origin that have superficial action. Among the most effective are the lipopeptide biosurfactants produced by Bacillus, especially surfactin. These biological products have a number of advantages that potentiate their applications, such as: stability to extreme conditions (pH, temperature), diversity of chemical structures, excellent surface and ecological properties, antibiotic action against pathogenic microorganisms, etc. In this context, the productions of total biosurfactants and Surfactin by Bacillus amyloliquefaciens IT45 were evaluated when the reagents concentration present in the culture medium varied. The submerged fermentations were carried out in an industrial shaker and in a pilot bioreactor of 40 liters capacity. In order to improve the reactants concentration, a Central Composite Design was developed to evaluate the influence of three variables (glucose syrup – Glucodry, yeast extract and calcium chloride) on superficial tension, total biosurfactant production and residual sugar. After statistical analyzes, when the variables were in the concentrations (g.L-1) of 20 for Glucodry, 15 for yeast extract and 4 for calcium chloride, the superficial tension (mN/m) reduces values above 50 to about 30, the residual sugar was minimal, around 31% and the total biosurfactant production was maximum, around 5.5 gL-1, after a period of 48 hours. The characterization of the biosurfactant identified Surfactin presence that was quantified in the retention time of 13.5 minutes. In order to know the real production of Surfactin and cellular biomass growth, fermentations were made in a 40 liter pilot bioreactor and the results were quite favorable. The most important culture medium suggested by the Central Composite Design, where glucose syrup, yeast extract and calcium chloride were in the concentrations (g.L-1) of 20, 15 and 4, respectively. For this fermentation, the cellular growth was 6.0 x 109 CFU.mL-1, Surfactin production was 0.63 g.L-1 and residual sugar was 28%. The results demonstrate that Bacillus amyloliquefaciens IT45 has potential to produce biosurfactants and does not require high concentrations of carbon and nitrogen sources for its development.
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Chu, Hoang Ha. "Identification and functions of type I signal peptidases of Bacillus amyloliquefaciens." [S.l. : s.n.], 2001. http://deposit.ddb.de/cgi-bin/dokserv?idn=964358999.

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Bashir, Abdallah. "Charakterisierung der Rolle von genereller Stressantwort und Sporulation bei der Anpassung von Bacillus-Stämmen an den Standort Boden." [S.l. : s.n.], 2004. http://archiv.ub.uni-marburg.de/diss/z2004/0512/.

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Mattapally, Peter Vijay. "Characterizing Bacillus amyloliquefaciens UCMB5113 on a Plant Model Arabidopsis thaliana." Thesis, Örebro universitet, Institutionen för naturvetenskap och teknik, 2014. http://urn.kb.se/resolve?urn=urn:nbn:se:oru:diva-38378.

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Organic farming is gaining importance and acceptance worldwide due to its beneficial effects in agriculture and standing against losses caused by chemical fertilizers, pesticides and fungicides. Plant growth promoting bacteria (PGPB) plays an important role in organic farming by fixing atmospheric nitrogen, chelate iron, solubilizing phosphorous, producing and modulating phytohormones, providing antibiotics against pathogens. Understanding interaction mechanisms between PGPB and plant will be helpful in developing new formulations to form a strong symbiotic relationship between plant and bacteria. Bacillus amyloliquefaciens UCMB5113 is a red pigmented, rod shaped Gram positive bacteria which has been isolated from fields of the Ukraine. In the present study UCMB5113 and its interactions with the plant has been characterized. There was a significant promotion of plant root growth and protection against biotic stress with the application of 10 μl of 1x107/ml CFU UCMB5113 culture in Arabidopsis. The UCMB5113 can significantly withstand plant antimicrobial activity to stimulate plant root growth, but needs root hair defective RHD proteins to stimulate root hair elongation. UCMB5113 has significantly inhibited primary root elongation and developed number of lateral roots and root hairs in ethylene over expressed mutant, which suggests that it may be affecting ethylene signaling pathway in plants. UCMB5113 has a distinct red pigmentation which is a 38.5kDa water soluble protein with maximum absorbance at 422nm. These features are similar to the Orange Carotenoid Protein (OCP) of Synechocystis PCC 6803. This red pigmented protein has no significant effect on plant root growth promotion. Further biochemical and molecular studies are required to characterize and confirm the mechanisms of interaction.

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Bonelli, Raquel Regina. "Extração e caracterização de uma substância antimicrobiana produzida por bacillus amyloliquefaciens." Florianópolis, SC, 2001. http://repositorio.ufsc.br/xmlui/handle/123456789/80287.

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Dissertação (mestrado) - Universidade Federal de Santa Catarina, Centro de Ciências Agrárias.
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Extraiu-se uma substância antimicrobiana de uma cultura 18-24 h de Bacillus amyloliquefaciens em Caldo Triptona de Soja suplementado com Extrato de Levedura a 0,6% (TSB-YE - OXOID) por precipitação com 20% de saturação de sulfato de amônio, ressuspensão em tampão fosfato pH 7,2, diálise em membrana 3,5kD (Spectra/Por) e esterilização por filtração (membrana 0,22mm - Millipore). Esta substância foi caracterizada segundo sua sensibilidade a temperatura (50, 75 e 100°C) e pH (2,0 a 9,0). Para determinação de sua natureza química empregaram-se as enzimas (Sigma) protease de S. griseus (P6911), protease de A. saitoi (P2143), a-quimotripsina (C4129) e pepsina (P6887) a 50mg/mL e a a-amilase Termamyl 120 L (Novo Nordisk) a 1% v/v. Para verificação da manutenção da atividade após tais ensaios, o extrato foi aplicado em poços preparados em placas de Ágar Triptona de Soja suplementado com Extrato de Levedura a 0,6% (TSA-YE - OXOID) pré-semeadas com 0,1 mL de uma cultura 105 UFC/mL de Listeria monocytogenes. Determinou-se sua concentração mínima inibitória (CMI) sobre uma cultura 104 UFC/mL de L. monocytogenes e seu peso molecular por eletroforese em gel de poliacrilamida (SDS-PAGE). Como resultado dos testes de caracterização, a substância apresentou-se estável a 50°C, perdendo sua atividade gradualmente a 75°C e rapidamente a 100°C. Também foi estável em valores de pH de 2,0 a 9,0. Perdeu parcialmente sua atividade quando tratada com as enzimas protease de S.griseus, protease de A. saitoi e a-amilase, indicando uma estrutura glicoproteica. A CMI foi 25% v/v e a substância agiu de forma bacteriostática. A eletroforese indicou um peso molecular inferior a 6.500 Daltons.

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Schulz, Denys. "Avaliação toxicológica do extrato bruto antibacteriano de Bacillus amyloliquefaciens R 10." Florianópolis, SC, 2008. http://repositorio.ufsc.br/xmlui/handle/123456789/91977.

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Tese (doutorado) - Universidade Federal de Santa Catarina, Centro de Ciências Agrárias. Programa de Pós-Graduação em Ciência dos Alimentos.
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Alguns microrganismos possuem a capacidade de produzir substâncias que podem influenciar no desenvolvimento de outros microrganismos. Desde os anos 50 é relatada a capacidade de várias espécies de bactérias do gênero Bacillus de produzir substâncias com atividade antimicrobiana como peptídeos, também denominados de bacteriocinas e enzimas como a subtilisina, subtilina, as proteases e as termolisinas. Muitos desses peptídeos e enzimas têm sido caracterizados bioquimicamente e geneticamente. Embora sejam conhecidas, a função estrutural, a biossíntese e modo de ação de alguns peptídeos antimicrobianos e enzimas, muitos aspectos desses compostos ainda permanecem desconhecidos. Frente a essa realidade, o presente estudo investigou a atividade antibacteriana e antifúngica e a toxicidade in vitro e in vivo do extrato bruto de Bacillus amyloliquefaciens R 10. O extrato bruto antibacteriano de Bacillus amyloliquefaciens R 10 foi obtido por técnicas de precipitação de proteínas, centrifugação, diálise e esterilização por filtração. A padronização das análises toxicológicas e de atividade antibacteriana e antifúngica foi realizada pela determinação da concentração de proteínas do extrato bruto de Bacillus amyloliquefaciens R 10. O ensaio de atividade antibacteriana e antifúngica foi realizado pela técnica de difusão em poços. Dentre os diferentes indicadores utilizaram-se bactérias Gram-positivas (Listeria monocytogenes NCTC 098630, Staphylococcus aureus ATCC 25923 e Enterococcus faecalis ATCC 29212), Gram-negativas (Escherichia coli ATCC 25922, Salmonella typhimurium ATCC 14028, Enterobacter aerogenes ATCC 13048 e Pseudomonas aeruginosa ATCC 9027), fungos (Aspergillus fumigatus ATCC 9197 e Penicillium commune J 238) e leveduras (Rhodotorula muscilaginosa J 350, Pichia anomala DSM 70255 e Kluveromyces marxianus ATCC 16045). O extrato bruto foi testado nas concentrações de 5, 20, 40, 60 e 80 g de proteínas, em ambos os ensaios de atividade antimicrobiana. Para avaliar a toxicidade do extrato bruto antibacteriano de Bacillus amyloliquefaciens R 10 foram utilizados ensaios in vitro (genotoxicidade) e in vivo (toxicidade aguda em ratos e camundongos e toxicidade de doses repetidas em camundongos). A investigação da genotoxicidade com células VERO ATCC-CCL 81 foi realizada pelo ensaio Cometa, utilizando-se 60, 80 e 100 g de proteínas do extrato bruto. Para análise toxicológica aguda em ratos Wistar (Rattus norvegicus), utilizou-se um grupo de 10 animais (5 machos e 5 fêmeas) tratado com o extrato bruto, por gavagem oral, com a dose de 2.000µg/kg (dose máxima recomendada). Os animais foram pesados no início e no final do experimento e observados por 14 dias quanto aos possíveis sinais de toxicidade (morte, coma, convulsão, prostração, ataxia, tremores, alteração na pele ou pêlo, alteração nas mucosas, diarréia e salivação). Para análise toxicológica aguda em camundongos Swiss (Mus musculus), utilizou-se o extrato bruto antibacteriano de Bacillus amyloliquefaciens R 10 por gavagem oral, numa única administração, nas doses de 5, 50, 500 e 5.000 g/kg a 80 camundongos (40 machos e 40 fêmeas, contendo 10 animais por grupo). Outros 20 camundongos (10 machos e 10 fêmeas) receberam veículo (salina, NaCl 0,9%), pela mesma via. Para análise toxicológica de doses repetidas em camundongos Swiss (Mus musculus) utilizou-se o extrato bruto antibacteriano de Bacillus amyloliquefaciens R 10 administrado por gavagem oral, por um período de 90 dias nas doses de 50, 500 e 5.000 g/kg em 120 animais (60 machos e 60 fêmeas, contendo 20 animais por grupo). Outros 40 camundongos (20 machos e 20 fêmeas) receberam veículo (salina, NaCl 0,9%), pela mesma via. Constatou-se nos ensaios de atividade antimicrobiana que o extrato bruto de Bacillus amyloliquefaciens R 10 não apresentou atividade contra os bolores e as leveduras nas concentrações testadas, e somente apresentou atividade antibacteriana frente Listeria monocytogenes NCTC 098630. O extrato bruto antibacteriano de Bacillus amyloliquefaciens R 10 não foi genotóxico para células VERO na concentração de 60µg/ml, porém apresentou genotoxicidade nas concentrações de 80 e 100µg/ml. Na análise toxicológica aguda em ratos a dose letal 50 (DL50) foi superior a 2.000µg/kg. Na análise toxicológica aguda em camundongos, o extrato bruto apresentou boa tolerabilidade e reduzidos efeitos tóxicos agudos importantes, com DL50 superior a 5.000µg/kg. Já na análise toxicológica de doses repetidas, a non toxic adverse effect level (NOAEL) para o extrato bruto antibacteriano de Bacillus amyloliquefaciens R 10 situa-se entre as doses de 50 e 500 g/kg. Os resultados indicam que o extrato bruto antibacteriano de Bacillus amyloliquefaciens R 10 apresenta um grande potencial como conservador natural de alimentos, dada sua marcante atividade antilisterial e sua relativa segurança toxicológica. Some microorganisms are able to produce substances that can influence the growth of other microorganisms. The ability of various bacterial species of the genus Bacillus to produce substances with antimicrobial activity such as peptides, also called bacteriocins, and enzymes such as subtilisin, subtilin, proteases and thermolysins has been reported since the 1950's. Many of these peptides and enzymes have been characterized biochemically and genetically. Although the structural function, biosynthesis and mode of action of some antimicrobial peptides and enzymes have been identified, many aspects of these compounds are still unknown. The present study investigated the antibacterial and antifungal activity and the in vitro and in vivo toxicity of the crude extract of Bacillus amyloliquefaciens R 10. The antibacterial crude extract of Bacillus amyloliquefaciens R 10 was obtained by techniques of protein precipitation, centrifugation, dialysis and sterilization through filtration. The standardization of the toxicological analysis, and antibacterial and antifungal activity was realized by determining the concentration of proteins in the crude extract of Bacillus amyloliquefaciens R 10. The assay of antibacterial and antifungal activity was done by agar diffusion technique in wells. Among the different indicators were used the Gram-positive bacteria (Listeria monocytogenes NCTC 098630, Staphylococcus aureus ATCC 25923 and Enterococcus faecalis ATCC 29212), Gram-negative (Escherichia coli ATCC 25922, Salmonella typhimurium ATCC 14028, Enterobacter aerogenes ATCC 13048 and Pseudomonas aeruginosa ATCC 9027), molds (Aspergillus fumigatus ATCC 9197 and Penicillium commune J 238), and yeasts (Rhodotorula muscilaginosa J 350, Pichia anomala DSM 70255 and Kluveromyces marxianus ATCC 16045). The crude extract was tested in concentrations of 5, 20, 40, 60 and 80 g of protein in both assays of antimicrobial activity. To evaluate the Bacillus amyloliquefaciens R 10 antibacterial crude extract's toxicity were used in vitro (genotoxicity) and in vivo (acute toxicity in rats and mice and repeated doses toxicity in mice) assays. The research of the genotoxicity with VERO ATCC-CCL 81 cells was made by Comet assay, using 60, 80 e 100 g of protein of the crude extract. To the acute toxicological analysis in rats Wistar (Rattus norvegicus), were used one group with 10 animals (5 males and 5 females) treated with the crude extract by oral gavage, with dose of 2,000µg/kg (maximum dose recommended). The animals were weighed at the beginning and at the end of the experiment, and observed during 14 days as the possible toxicity signs (death, coma, convulsions, prostration, ataxy, tremors, skin or hair alterations, mucous alteration, diarrhea and salivation). To the acute toxicological analysis in Swiss mice (Mus musculus), the antibacterial crude extract of Bacillus amyloliquefaciens R 10 was used by oral gavage, in a single administration, in doses of 5, 50, 500 and 5,000 g/kg to 80 mice (40 males and 40 females, with 10 animals per group). Other 20 mices (10 males and 10 females) received the vehicle (saline, NaCl 0.9%), through the same route. To the repeated doses toxicological analysis in Swiss mice (Mus musculus) the antibacterial crude extract of Bacillus amyloliquefaciens R 10 was administrated by oral gavage during 90 days in doses of 50, 500 and 5,000 g/kg in 120 animals (60 males and 60 females, in groups of 20 animals). Other 40 mice (20 males and 20 females) received the vehicle (saline, NaCl 0.9%), through the same route. It was verified in the antimicrobial activity assays that the crude extract of Bacillus amyloliquefaciens R 10 didn't present activity against molds and yeasts in the tested concentrations, and only presented antibacterial activity against Listeria monocytogenes NCTC 098630. The antibacterial crude extract of Bacillus amyloliquefaciens R 10 was not genotoxic to VERO cells at concentration of 60µg/ml, but showed genotoxicity at concentration of 80 and 100µg/ml. In the acute toxicological analysis in rats the letal dose 50 (DL50) was higher than 2,000µg/kg. In the acute toxicological analysis in mice, the crude extract presented good tolerability and litle important acute toxic effects with DL50 higher than 5,000µg/kg. In the repeated doses toxicological analysis, a non toxic adverse effect level (NOAEL) for the antibacterial crude extract is between the doses of 50 and 500 g/kg. The results showed that the antibacterial crude extract of Bacillus amyloliquefaciens R 10 presents a great potential as a natural food preservative, due to its notable antilisterial activity, and its relative toxicological security.

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Scholz, Romy. "Synthese der Bacteriocine Amylocyclicin A und Plantazolicin in Bacillus amyloliquefaciens FZB42." Doctoral thesis, Humboldt-Universität zu Berlin, Mathematisch-Naturwissenschaftliche Fakultät I, 2011. http://dx.doi.org/10.18452/16283.

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Bacillus amyloliquefaciens FZB42 ist ein grampositives Bodenbakterium. Es kann in der Rhizosphäre das Wachstum von Pflanzen fördern und durch die Produktion von Sekundärmetaboliten phytopathogene Organismen hemmen. Aus der Genomanalyse und den dazugehörigen Arbeiten war bekannt, dass Bacillus amyloliquefaciens FZB42 nicht-ribosomal je drei antimikrobielle Polyketide und Lipopeptide herstellt, sowie zwei Siderophore und das Dipeptid Bacilysin. Für Bacillus typische Lantibiotika oder große Bacteriocine wurden nicht gefunden. In dieser Arbeit wird erstmalig gezeigt, dass Bacillus amyloliquefaciens FZB42 auf ribosomale Weise antibakterielle Peptide herstellt. Zwei bisher unbekannte Bacteriocine, Amylocyclicin A und Plantazolicin, und deren dazugehörigen Gencluster konnten identifiziert und charakterisiert werden. Amylocyclicin A ist ein unmodifiziertes Peptid, dessen N- und C-Terminus kovalent verbunden sind. Es wurde der Gruppe I der zirkulären Bacteriocine zugeordnet, dessen Mitglieder sich durch schwache Homologie untereinander, aber durch wahrscheinlich ähnliche 3D-Strukturen auszeichnen. Die Masse beträgt 6381 Da und die Substanz ist stark aktiv gegen grampositive Bakterien. Das Biosynthesecluster umfasst sechs Gene für die Synthese, den Export, die Zyklisierung und die Immunität. Plantazolicin ist ein hydrophobes, stark modifiziertes Peptid aus der TOMM-Gruppe, einer Gruppe aus Microcin B17-ähnlichen Peptiden, die nach neueren Erkenntnissen verbreiteter ist, als bisher bekannt. Plantazolicin ist schwach aktiv gegen grampositive Bakterien und besitzt die Masse 1335 Da. Das Biosynthesecluster umfasst zwölf Gene, mit allen nötigen Genen für Synthese, Modifikation, Regulation, Immunität und Export.
Bacillus amyloliquefaciens FZB42 is a Gram-positive, plant-associated bacterium, which stimulates plant growth and produces secondary metabolites that suppress soil-borne plant pathogens. Five gene clusters direct the non-ribosomal synthesis of the cyclic lipopeptides surfactin, bacillomycin, fengycin, an unknown peptide and the iron-siderophore bacillibactin. Three gene clusters direct the non-ribosomal synthesis of the antibacterial acting polyketides macrolactin, bacillaene and difficidin; in addition to the non-ribosomal synthesis of the antibacterial dipeptide bacilysin. Genes involved in ribosome-dependent synthesis of lantibiotics and other peptides are scarce. Only two incomplete gene clusters directing immunity against mersacidin and subtilin were found. In this work two ribosomally synthesized antibacterial peptides, amylocyclicin A and plantazolicin, and their corresponding gene clusters were identified. Amylocyclicin A is a circular peptide with a mass of 6381 Da and strong activity against Gram-positive bacteria. Six genes are responsible for the synthesis, maturation, export and immunity of this peptide belonging to group I of circular bacteriocins. Plantazolicin is a strongly modified hydrophobic peptide bearing a molecular mass of 1,335 Da and displaying antibacterial activity toward closely related Gram-positive bacteria. Essential modification contains the incorporation of azole heterocycles, which derive from Cys, Ser, and Thr residues of the precursor peptide and addition of two methyl groups. Twelve genes are responsible for synthesis, modification, export and immunity of this peptide belonging to the TOMM group of thiazol/oxazol modified microcins.

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Benitez, Lisianne Brittes. "Caracterização de peptídeos antimicrobianos de Bacillus amyloliquefaciens com atividade antibacteriana, antifúngica e amebicida." reponame:Biblioteca Digital de Teses e Dissertações da UFRGS, 2010. http://hdl.handle.net/10183/29950.

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Bacteriocinas são peptídeos pequenos sintetizados por bactérias que possuem propriedades antimicrobianas. O presente estudo teve por objetivos identificar e caracterizar uma bacteriocina produzida por Bacillus amyloliquefaciens LBM 5006, isolado de solo da Mata Atlântica, Santa Catarina, Brasil. A produção da substância antimicrobiana tem início na fase exponencial de crescimento e sua atividade máxima ocorre na fase estacionária. O efeito do cocultivo de B. amyloliquefaciens com diferentes bactérias na produção do antimicrobiano foi investigado. O cultivo de células de Escherichia coli intactas ou inativadas pelo calor com a cepa produtora aumentou a síntese da substância antimicrobiana. A bacteriocina LBM 5006 apresentou amplo espectro de ação, produzindo atividade antagonista contra bactérias, fungos e amebas patogênicas. O modo de ação contra células de Listeria monocytogenes e Paenibacillus larvae foi bactericida e bacteriolítico. Atividade esporicida foi observada contra esporos de P. larvae após tratamento com 1600 UA mL-1. Efeitos amebistático e amebicida contra trofozoítos de Acanthamoeba polyphaga com a consequente lise celular foram observados. A bacteriocina não teve efeito inibitório em células Vero nas concentrações que foram efetivas contra as amebas. A substância antimicrobiana foi isolada por precipitação com sulfato de amônio, cromatografia de gel filtração e extração com 1-butanol. O espectro ultravioleta foi típico de um polipeptídeo e o infravermelho indicou a presença de ligações peptídicas e grupamentos acil na sua estrutura. Análises por espectroscopia de massas indicaram que B. amyloquefaciens LBM 5006 produziu dois peptídeos antimicrobianos, com 1058 Da e 1464 Da, correspondentes a peptídeos tipo-iturina e tipo-fengicina, respectivamente.
Bacteriocins are small peptides synthesized by bacteria which have antimicrobiall properties. This study aimed to identify and characterize a bacteriocin produced by Bacillus amyloliquefaciens LBM 5006, isolated from soil of Mata Atlântica, Santa Catarina, Brazil. It has been observed that the antimicrobial substance production starting at the exponencial grown phase and maximum activity occur at stationary phase. The effect of different bacteria on the production of antimicrobial activity by Bacillus amyloliquefaciens LBM 5006 was investigate. It was concluded that the presence of intact or thermally inactivated cells of Escherichia coli enhanced the synthesis of antimicrobial peptides by B. amyloliquefaciens strain. Bacteriocin LBM 5006 showed broad spectrum of action, producing antagonistic activity against bacteria, fungi and pathogenic amoebas. The mode of action against Listeria monocytogenes and Paenibacillus larvae was bactericidal and bacteriolytic. Sporicidal activity was observed against P. larvae spores after treatment whit 1600 AU mL-1. Amoebicidal and amoebistatic effects were detected against throphozoites of Acanthamoeba poliphaga and cell lysis. The bacteriocin had no inhibitory effect on Vero cells at concentrations that were effective against amoebas.The antimicrobial substance was isolated by ammonium sulfate precipitation, gel filtration chromatography and 1-butanol extraction.The ultraviolet spectrum was typical of a polypeptide and the infrared spectrum indicates the presence of peptide bonds and acyl group(s) in its structure. Mass spectroscopy analysis indicated that B. amyloliquefaciens LBM 5006 produces two antimicrobial peptides, with main peaks at m/z 1058 Da and 1464 Da, corresponding to iturinlike and fengycin-like peptides, respectively.

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Larini, Mariana Munhoz. "Produção de surfactina por Bacillus amyloliquefaciens MO.04b em mistura de resíduos agroindustriais." Universidade Estadual de Londrina. Centro de Ciências Exatas. Programa de Pós-Graduação em Biotecnologia, 2017. http://www.bibliotecadigital.uel.br/document/?code=vtls000216058.

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O Brasil, importante no cenário agrícola mundial, é produtor de soja, grão de alto teor proteico, cana de açúcar e arroz. Essas culturas, quando processadas, geram materiais que são potenciais substratos para o cultivo microbiano e produção de metabólitos de alto valor agregado. A fermentação em estado sólido (FES) possibilita o cultivo de microrganismos em condição de baixo teor de umidade e favorece o aproveitamento de diferentes resíduos agroindustriais. O objetivo deste trabalho foi avaliar a produção de surfactina, um biossurfctante lipopeptídico, por FES, utilizando o Bacillus amyloliquefaciens MO.04b. O microrganismo foi cultivado em mistura de resíduos agroindustriais contendo farelo de soja, bagaço de cana-de-açúcar e casca de arroz, umedecidos com solução de sais acrescida de glicose 0,2 % (m/v), inoculados com suspensão de células (107 - 108 UFC mL-1) e incubados a 37 ± 2 °C. Após a interrupção da FES com água destilada, a mistura foi homogeneizada, a biomassa determinada por turbidimetria a 600 nm e contagem das unidades formadoras de colônias (UFC). Essa mistura foi submetida a centrifugação a 3.500 x g por 15 minutos. O peso do material sólido fermentado foi determinado por gravimentria e o extrato livre de células (ELC) utilizado para a determinação do pH (inicial e final), da atividade das enzimas proteases, xilanases, celulases e lacases, das proteínas totais e dos açúcares totais e redutores. Os lipipeptídeos contidos no ELC foram precipitados com adição de HCl 6 mol L-1 até pH 2,0 e mantido em repouso durante 12 horas e extraídos com a mistura de clorofórmio e metanol [CHCl3:CH3OH (4:1)]. O material obtido foi então liofifizado. A quantificação de surfactina foi realizada por cromatografia líquida de alta eficiência (CLAE). O pH final dos cultivos foi crescente ao longo do tempo de cultivo e atingiu o máximo de 8,5 em 72 horas, assim como a biomassa, que cresceu ao longo do tempo, até 18 24 horas, atingindo o máximo de 1 x 108 UFC mL-1. Quanto ao material sólido fermentado, foi verificada a redução progressiva do peso seco, de 1,2207g (0 horas) até 0,9268 g (72 horas), mostrando que em 72 horas de cultivo o microrganismo hidrolisou 25 % do total dos resíduos agroindustriais. A atividade de proteases e xilanases foram 0,946 U mL-1 e 0,81 U mL-1, respectivamente, entre 6 e 12 horas de cultivo. Não foram detectadas atividades de celulase e lacase. A surfactina foi produzida em todos os períodos analisados, e teve maior produção em 18 horas de cultivo, de 64,15 mg mL-1. A mistura de resíduos utilizada favoreceu o crescimento do microrganismo, o qual foi capaz de consumir 25 % dos resíduos nas condições de FES apresentadas. Tais condições propiciaram à bactéria a produção das enzimas protease e xilanase e de surfactina, cuja maior produção ocorreu em 18 horas de cultivo.
Brazil, important in the world's agricultural scenario, is a producer of soybeans, high-protein grain, sugar cane and rice. These cultures, when processed, generate materials that are potential substrates for the microbial culture and production of metabolites of high added value. A solid state fermentation (FES) allows the cultivation of microorganisms in a condition of low moisture content and favors the use of different agroindustrial residues. The objective of this work is to evaluate the production of surfactin, a lipopeptide biosurfactant, by FES using Bacillus amyloliquefaciens MO.04b. The microorganism was cultivated in a mixture of agroindustrial residues containing soybean meal, sugarcane bagasse and rice husk, moistened with salt solution plus 0.2% glucose (m/v), inoculated with cell suspension (107-108 CFU ml-1) and incubated at 37 ± 2 °C. After interruption of FES with distilled water, the mixture was homogenized, the biomass determined by turbidimetry at 600 nm and counting of the colony forming units (CFU). This mixture was subjected to centrifugation at 3500 x g for 15 min. The weight of the fermented solid material was determined by gravimetry and the free cell extract (ELC) used for the determination of the pH (initial and final), activity of enzymes protease, xylanases, cellulases and laccases, total proteins, total sugars and reduced sugar. The lipids contained in the ELC were precipitated with addition of 6 mol L-1 HCl to pH 2.0 and held for 12 hours and extracted with a mixture of chloroform and methanol [CHCl3:CH3OH (4:1)]. The obtained material was then lyophilized. The quantification of surfactin was performed by high performance liquid chromatography (HPLC). The final pH of the cultures was increased over the growing time and reached the maximum of 8.5 in 72 hours, as well as the biomass, which grew over time, up to 18 - 24 hours, reaching a maximum of 1 x 108 CFU mL-1. As regards the fermented solid material, a progressive reduction of dry weight was observed from 1.2207g (0 h) to 0.9268 g (72 h), showing that in 72 h of culture the microorganism hydrolyzed 25% of the total agroindustrial waste. The activity of proteases and xylanases were 0.946 U mL-1 and 0.81 U mL-1, respectively, between 6 and 12 hours of culture. No cellulase and laccase activities were detected. Surfactin was produced in all analyzed periods, and had a higher production in 18 hours of culture, of 64.15 mg mL-1. The mixture of residues used favored the growth of the microorganism, which was able to consume 25% of the residues under the FES conditions presented. These conditions allowed the bacterium to produce protease and xylanase and surfactin enzymes, whose highest production occurred in 18 hours of culture.

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Mariappan, Aruljothi. "Molecular mechanisms controlling bacilysin biosynthesis in plant growth promoting rhizobacterium - Bacillus amyloliquefaciens FZB42." Doctoral thesis, Humboldt-Universität zu Berlin, Mathematisch-Naturwissenschaftliche Fakultät I, 2012. http://dx.doi.org/10.18452/16554.

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Bacillus amyloliquefaciens FZB42 ist ein grampositives Bakterium, das in der Rhizosphäre das Pflanzenwachstum fördert (PGPR - Plant Growth Promotion) und pathogene Organismen hemmt. Abgesehen von dieser Fähigkeit produziert es eine Vielzahl von sekundären Metaboliten, die sowohl ribosomale als auch nicht-ribosomale Peptide umfassen. In dieser Arbeit erfolgte die Untersuchung der transkriptionellen Aktivierung und Regulation der Bacilysin- Biosynthese an den Promotoren der bac- und ywfH- Gene. Durch 5´-Deletionsanalysen wurde der Promotor von Bacilysin identifiziert. Die A (Sigmafaktor A) - abhängige Transkription startet über die konservierten Promotorelemente (-10 und -35) von den bac- und ywfH Genen. Die Untersuchungen der Promotoraktivitäten vom Wildtyp und den erzeugten Regulationsmutanten erfolgten über in vivo ß-Galaktosidase-(Reporter)-Assays. Die Ergebnisse der Reporter-Aktivitäten zeigten, dass Transkriptionsregulatoren die Expression der Bacilysin- Gene aktivieren. Mehrere globale Regulatoren wie DegU, ComA, Hpr und AbrB beeinflussen die Genexpression. In dieser Arbeit wurde mithilfe von DNaseI Footprinting-Analysen die DegU-Bindung an die bac- und ywfH- Promotoren bestätigt.Die negative Regulation der Bacilysin-Biosynthese wird durch den Regulator der transienten Phase Hpr bewerkstelligt. Eine direkte Hpr-Bindung an bac Promotor wurde mit DNaseI Footprint-Analysen gezeigt. Der Promotor des monocistronischen Gens ywfH wurde aber durch Hpr nicht beeinflusst. Die anderen Transkriptionsregulatoren, wie ComA und AbrB, regulieren die Genexpression von Bacilysin indirekt über DegQ und Hpr. In dieser Arbeit konnte demonstriert werden, dass der globale Regulator AbrB den Promotor vom hpr-Gen direkt kontrolliert. Zusammenfassend liefert diese Studie neue Informationen über die genetische Regulation der Bacilysin- Biosynthese in B. amyloliquefaciens FZB42.
Bacillus amyloliquefaciens FZB42 is a Gram-positive, pathogen-suppressing and plant-growth promoting rhizobacterium. Apart from this ability, it produces a vast array of secondary metabolites, which includes both ribosomal and non-ribosomal peptides. In this work, the transcriptional activation and regulation of bacilysin biosynthesis were studied at the promoters of bac and ywfH genes. The promoter of bacilysin was identified using 5''-deletion analysis. Sigma factor A (σA) was found to start transcription via conserved promoter elements (-10 and -35) of bac and ywfH genes. lacZ reporter fusion studies were performed in wild type and regulatory mutants. The results show the involvement of transcriptional regulators to activate the expression of bacilysin genes. Several global regulators such as DegU, ComA, Hpr and AbrB were identified and found to influence gene expression. In particular, I confirmed DegU binding in bac and ywfH promoters using radioactive DNase I footprinting. Furthermore, Hpr, a transition state regulator was found negatively to control bacilysin biosynthesis. Hpr binding to bac promoter was demonstrated using radioactive DNase I footprinting. Remarkably, Hpr does not influence the promoter of the monocistronic gene, ywfH. The other transcriptional regulators, such as ComA and AbrB, were correlated indirectly to affect the gene expression of bacilysin via DegQ and Hpr, respectively. The gene regulation of hpr was studied in this work. It was demonstrated that AbrB, a global regulator, directly controls the promoter of the hpr gene. However, the consensus sequence for AbrB binding was not identified, since it covers the entire promoter region in the DNA-protein interaction study. To conclude, this study provides new information regarding the genetic regulation of bacilysin biosynthesis in B. amyloliquefaciens FZB42.

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Koumoutsi, Alexandra. "Functional genome analysis of the plant-growth promoting bacterium Bacillus amyloliquefaciens strain FZB42." Doctoral thesis, Humboldt-Universität zu Berlin, Mathematisch-Naturwissenschaftliche Fakultät I, 2007. http://dx.doi.org/10.18452/15586.

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Bacillus amyloliquefaciens FZB42 ist ein im Boden weit verbreitetes Bakterium. Es kolonisiert Pflanzenwurzeln und werde als Biodünger verwendet, da sie in der Lage sind, Pflanzenwachstum zu fördern. Der domestizierte Stamm B. subtilis 168 ist eng verwand mit B. amyloliquefaciens FZB42, fördert jedoch kein Pflanzenwachstum. Als ein erster Ansatz zur Ermittlung von Gendifferenzen zwischen FZB42 und B. subtilis 168 - wobei zum damaligen Zeitpunkt nur die Genomsequenz letzteren Organismus bekannt war - wurde die Supression Subtractive Hybridisation (SSH) angewandt. Hierdurch wurden mehrere einzigartige Gene in B. amyloliquefaziens identifiziert. Unterdessen beteiligte sich unser Labor in Kollaboration mit dem GenoMik Network in Göttingen an einem Projekt, dessen Ziel die komplette Sequenzierung des Genoms von B. amyloliquefaciens war. Der Hauptanteil der Arbeit, sowie die Koordination des gesamten Projekts wurden von Xiao-Hua Chen und mir selbst durchgeführt. B. amyloliquefaciens FZB42 besitzt die srf, fen, pks1 (bae), bac und dhb Operons, welche für die Synthese von Surfactin, Fengycin, Bacillaene, Bacilysin und Bacillibactin verantwortlich sind und die ebenfalls im Genom von B. subtilis 168 enthalten sind. Das Genom von B. amyloliquefaciens FZB42, beinhaltet die bmy Gencluster, die die Synthese von Bacillomycin D kontrolliert. Ein weiteres in dieser Arbeit verfolgtes Ziel war die Identifizierung der regulatorischen Wege, die die Expression von Bacillomycin D steuern. Es wurde gezeigt, dass globale Regulatoren, wie beispielsweise DegU, DegQ und ComA, die alternativen Sigmafaktoren sigB und sigH und ein neuartiges Rap-Protein die transkriptionale Aktivität des in dieser Arbeit identifizierten Hauptpromotors des bmy-Operons beeinflussen. Es wurde gezeigt, dass DegU seine Effekte nach direkter Bindung an zwei unterschiedliche Regionen im bmy-Promotor ausübt. Es wurde außerdem gezeigt, dass DegU abgesehen von der Aktivierung des Hauptpromoters des bmy-Operons eine zusätzliche, vermutlich eine post-transkriptionale Rolle spielt. Auf ähnliche Weise erwies sich YczE, ein Membranprotein unbekannter Funktion, das neben sfp kodiert liegt, als essentiell für die Bacillomycin D-Produktion. Der Effekt wurde auf einem post-transkriptionalen Level ausgeübt und war unabhängig von DegU.
Bacillus amyloliquefaciens FZB42 is widely distributed in the soil. It colonizes the plant roots and is used as bio-fertilizer, since they can promote plant growth.The domesticated strain of B. subtilis 168 is closely related to B. amyloliquefaciens FZB42, but does not promote plant growth. As a first approach to detect gene differentiation between FZB42 and B. subtilis 168, and since only the genome sequence of the latter was known at that point, Suppression Subtractive Hybridization (SSH) was employed. Thereby, several unique genes of B. amyloliquefaciens FZB42 could be identified. Meanwhile, our laboratory became engaged in a project aiming to define the complete genome sequence of B. amyloliquefaciens FZB42, in collaboration with the GenoMik Network in Göttingen. The major part of the work and the co-ordination of the whole process were performed by Xiao-Hua Chen and myself. B. amyloliquefaciens FZB42 possesses the srf, fen, pks1 (bae), bac and dhb operons, which are also shared by B. subtilis 168. In addition, the genome of B. amyloliquefaciens FZB42 contains the bmy gene clusters, which controls the synthesis of bacillomycin D. A further issue pursued in this work was to identify the regulatory pathways that govern the expression of bacillomycin D. Global regulators, such as DegU, DegQ and ComA, the alternative sigma factors, sigB and sigH, and a novel Rap protein were found to affect the transcriptional activation of the main promoter of the bmy operon identified in this work. In particular, DegU was shown to mediate its effects, after binding directly to two sites at the bmy promoter region. DegU was shown to play an additional role on bacillomycin D production, presumably a post-transcriptional one. Similarly, YczE, a membrane protein of unknown function, encoded adjacently to sfp proved to be essential for bacillomycin D production, but dispensable for the production of the rest peptide antibiotics produced by B. amyloliquefaciens FZB42. The effect was mediated at a post-transcriptional level and was independent of DegU.

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Lisbôa, Márcia Pagno. "Caracterização de um peptídeo antimicrobiano produzido por linhagem de Bacillus amyloliquefaciens isolada de solo." reponame:Biblioteca Digital de Teses e Dissertações da UFRGS, 2006. http://hdl.handle.net/10183/7475.

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Em anos recentes, surgiram numerosos casos de intoxicação alimentar envolvendo patógenos emergentes. Estes casos levaram a um aumento da preocupação com a preservação dos alimentos minimamente processados e com a segurança alimentar. Este fato está induzindo a pesquisa por inibidores para estes patógenos e fatores para prolongar a vida de prateleira de produtos alimentícios. Entre as novas alternativas na preservação está a utilização de peptídeos antimicrobianos produzidos por bactérias. No presente trabalho uma bactéria identificada como Bacillus amyloliquefaciens LBM 5006 isolada de solos de mata Atlântica de Santa Catarina foi selecionada dentre outros microrganismos e sua capacidade de produzir antimicrobianos foi avaliada. O extrato bruto da cultura do isolado LBM 5006 foi caracterizado, sendo ativo contra importantes bactérias patogênicas e deteriorantes como Listeria monocytogenes, Bacillus cereus, Erwinia carotovora, Escherichia coli, dentre outras. Houve maior produção do antimicrobiano quando a bactéria foi propagada em caldo infusão de cérebro e coração (BHI) a 37o C durante 48 h. Após concentração, a atividade antimicrobiana resistiu ao tratamento com enzimas proteolíticas. A atividade antimicrobiana foi verificada em pHs ácidos, sendo inibida em pH 9 e 10. O extrato foi purificado por meio de cromatografia de gel filtração e extração com butanol. O teste qualitativo de ninidrina, juntamente com a espectroscopia de infravermelho e ultravioleta, feitos com a substância purificada revelou que o antimicrobiano possui natureza protéica. O antimicrobiano apresentou um efeito bacteriostático contra 106 UFC/mL de Listeria monocytogenes na concentração de 25 AU/ml.
In recent years, an increase in food poisoning cases involving emerging pathogens have been described. These cases have increased the preoccupation with the preservation of minimally processed food products and food safety. This fact is leading the research for inhibitory substances and for factors to extend the shelflife of food products. Among the new preservation alternatives is the utilization of antimicrobial peptides produced by bacteria. In present work a bacterium identified as Bacillus amyloliquefaciens LBM 5006 isolated from Atlantic Forest soils of Santa Catarina was selected among other microorganisms and its capacity to produce antimicrobials was evaluated. The crude culture supernatant of the strain LBM 5006 was characterized, being active against important pathogenic and spoilage bacteria like Listeria monocytogenes, B. cereus, E. carotovora, E. coli, among other. Maximum production of antimicrobial activity was observed when the bacteria was propagated in Brain Heart Infusion (BHI) at 37o C for 48 h. After concentration, the antimicrobial activity was resistant to proteolytic enzymes. Antimicrobial activity was verified in acid pHs, but it was lost at pH 9 and 10. The supernatant was purfied by gel filtration chromatography and butanol extraction. The qualitative ninhydrin test, together with infrared and ultraviolet spectroscopy of purified substance revealed that the antimicrobial has a peptide moiety. A dose of 25 UA/mL caused a bacteriostatic effect against 106 UFC/mL Listeria monocytogenes.

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Oliveira, Marllon José Karpeggiane de. "Avaliação de probiótico comercial como alternativa aos antibióticos promotores de crescimento, utilizando um modelo de desafio sanitário em frangos de corte /." Jaboticabal, 2019. http://hdl.handle.net/11449/182523.

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Orientador: Nilva Kazue Sakomura
Resumo: Doenças entéricas se apresentam como um dos maiores problemas da produção avícola. Historicamente, antibióticos promotores de crescimento (APC’s) têm sido usados para controlar essa situação. No entanto, as preocupações a respeito de resistência bacteriana a antibióticos têm levado a proibições regionais para reduzir o uso de APC’s. Alternativamente, microrganismos administrados diretamente na dieta como por exemplo os probióticos, podem impactar positivamente sobre a microbiota intestinal, prevenindo problemas entéricos que, inevitavelmente culminam em perdas no desempenho das aves. Outro ponto importante a ser considerado quando se fala em desafio sanitário é o consumo de ração dos animais. A utilização de modelos matemáticos que consideram o fator desafio para modelagem do consumo ainda é escasso. Nesse sentido, essa dissertação foi conduzida com dois principais propósitos: 1) investigar os efeitos de Bacillus amyloliquefaciens CECT 5940 como probiótico (DFM) sozinho ou em associação com bacitracina metileno disalicilato (BMD) em frangos de corte sobre desafio de patógeno entérico. 2) modelar o consumo relativo de animais sobre condição de desafio sanitário e integrar esse modelo dentro de um modelo mecanicista de simulação, o Broiler Growth Model (BGM). Assim, permitindo a modelagem do desempenho de animais sob condição de desafio sanitário. Para isso, um total de 1.530 frangos de corte machos Cobb500 com um dia de idade foram casualizados em cinco tratamentos, com nove r... (Resumo completo, clicar acesso eletrônico abaixo)
Abstract: Enteric diseases appear one of the biggest problems of poultry production. Historically, antibiotics growth promoter (AGPs) have been used to control this framework. However, concerns about antibiotic resistance bacteria have led regional bans for reducing the use of AGPs. Alternatively, microorganisms directly supplemented in diets such as probiotics, may exhibit a positive impact on intestinal microbiota avoiding enteric problems that inevitably causes loss on broilers performance. Another important point to be considered when talking about sanitary challenge is the feed intake. The use of mathematical models that consider the challenge factor for modelling in this feed intake is still scarce. In this sense, this project was conducted with two main purposes: 1) to investigate the effects of Bacillus amyloliquefaciens CECT 5940 as a probiotic (DFM) alone or in combination with bacitracin methylene disalicylate (BMD) in broilers on enteric pathogen challenge. 2) Model the relative feed intake of animals under sanitary challenge condition and to integrate this model into the mechanistic simulation model, the Broiler Growth Model (BGM). Thus, allowing the modelling of performance of animals under sanitary challenge condition. For such, a total of 1,530-day-old male Cobb500 chicks, were randomly assigned to five treatments, with nine replicate pens with 34 birds each. Treatments included positive control (PC, basal diet without additives or challenge); negative control (NC, basa... (Complete abstract click electronic access below)
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Watson, J. S. "Effect of fermentation conditions on release of intracellular material from Bacillus amyloliquefaciens by autolysis." Thesis, Teesside University, 1986. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.372856.

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Petrillo, Thalita Regina. "Efeito probiótico do Bacillus amyloliquefaciens na aerocistite aguda em tilápias-do-Nilo (Oreochromis niloticus) /." Jaboticabal, 2015. http://hdl.handle.net/11449/154734.

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Orientador: Julieta Rodini Egracia de Moraes
Banca: Rogério Salvador
Banca: Fernanda Nogueira Valentin
Banca: Sérgio Henrique Canello Schalch
Banca: João Batista Kochenborger Fernandes
Resumo: A utilização de probióticos surge como uma alternativa, para o uso de antibióticos em peixes, evitando que os mesmos adoeçam. O escopo deste trabalho foi avaliar os efeitos da alimentação adicionada de probiótico contendo Bacillus amyloliquefaciens na inflamação aguda contra A.hydrophila em tilápias-do-Nilo (Oreochromis niloticus). Para tanto foram utilizados 936 exemplares de tilápias-do-Nilo (30g), distribuídas uniformemente em 12 tanques-rede sendo 78 peixes em cada. Os peixes foram alimentados durante 60 dias com ração adicionada com a bactéria probiótica em diferentes concentrações, sendo constituídos os seguintes grupos; Grupo 1 (controle) alimentado com ração basal sem adição de probiótico: Grupo 2 (T1) suplementados com 0,75 gramas de probiótico/kg de ração; Grupo 3 (T2) suplementados com 1,5 gramas de probiótico/kg de ração; Grupo 4 (T3) suplementados com 3 gramas de probiótico/kg de ração. Para o estímulo inflamatório inoculou-se na bexiga natatória dos peixes 0,5 ml de solução salina 0,65% no grupo controle e 0,5 ml x106 de uma solução, contendo A. hydrophila inativada nos demais grupos. As coletas de sangue e exsudato inflamatório foram realizadas nos tempos de 12, 24 e 48 horas após estímulo inflamatório (HPE), para as seguintes análises: variáveis hematológicas, índices de glicose basal, atividade respiratória dos leucócitos sanguíneos (burst oxidativo), características qualitativas e quantitativas do exsudato, atividade lítica das proteases do soro e atividade de aglutinação bacteriana. Os resultados foram comparados por meio de análise de variância (ANOVA) ao nível de 5% de probabilidade e a diferença entre as médias foi comparada pelo teste de Tukey. Não foram observadas variações hematológicas significativas entre os grupos de tilápias-do-Nilo tratadas e estimuladas e o grupo controle, o mesmo pode ser observados...
Abstract: The use of probiotics arises as an alternative to the use of antibiotics in fishes, preventing them from becoming sick. The scope of this study was to evaluate the effects of food with added probiotic containing Bacillus amyloliquefaciens in acute inflammation against A.hydrophila in the Nile-tilapia (Oreochromis niloticus). To that end, there were used 936 copies of the Nile-tilapia (30g), evenly distributed in 12 net tanks with 78 fish in each. The fishes were fed for 60 days with food added with the probiotic bacteria in different concentrations, with the following groups being formed; Group 1 (control) fed with basal food without added probiotic: Group 2 (T1) supplemented with 0.75 grams of probiotic / kg food; Group 3 (T2) supplemented with 1.5 grams of the probiotic / kg food; Group 4 (T3) supplemented with 3 grams of probiotic / kg food. For the inflammatory stimulus it was inoculated in the swimming bladder of fish 0.5 ml of saline solution 0.65% in the control group and 0.5 ml x106 of a solution containing inactivated A. hydrophila in the other groups. Blood collections and inflammatory exudate were held in the periods of 12, 24 and 48 hours after the inflammatory stimulus (HPE), for the following analysis: haematological variables, basal glucose levels, respiratory activity of sanguine leukocytes (oxidative burst), qualitative and quantitative characteristics of the exudate, lytic activity of serum proteases and agglutination bacterial activity. The results were compared using analysis of variance (ANOVA) at 5% probability and the difference between the means was compared by Tukey test. There were no significant hematological changes between the groups of Nile-tilapia treated and stimulated and the control group, the same thing can be observed in glycemic indeces and respiratory activity of blood leucocytes. The inflammatory exudate was characterized by the predominant accumulation ...
Doutor

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Petrillo, Thalita Regina [UNESP]. "Efeito probiótico do Bacillus amyloliquefaciens na aerocistite aguda em tilápias-do-Nilo (Oreochromis niloticus)." Universidade Estadual Paulista (UNESP), 2015. http://hdl.handle.net/11449/154734.

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A utilização de probióticos surge como uma alternativa, para o uso de antibióticos em peixes, evitando que os mesmos adoeçam. O escopo deste trabalho foi avaliar os efeitos da alimentação adicionada de probiótico contendo Bacillus amyloliquefaciens na inflamação aguda contra A.hydrophila em tilápias-do-Nilo (Oreochromis niloticus). Para tanto foram utilizados 936 exemplares de tilápias-do-Nilo (30g), distribuídas uniformemente em 12 tanques-rede sendo 78 peixes em cada. Os peixes foram alimentados durante 60 dias com ração adicionada com a bactéria probiótica em diferentes concentrações, sendo constituídos os seguintes grupos; Grupo 1 (controle) alimentado com ração basal sem adição de probiótico: Grupo 2 (T1) suplementados com 0,75 gramas de probiótico/kg de ração; Grupo 3 (T2) suplementados com 1,5 gramas de probiótico/kg de ração; Grupo 4 (T3) suplementados com 3 gramas de probiótico/kg de ração. Para o estímulo inflamatório inoculou-se na bexiga natatória dos peixes 0,5 ml de solução salina 0,65% no grupo controle e 0,5 ml x106 de uma solução, contendo A. hydrophila inativada nos demais grupos. As coletas de sangue e exsudato inflamatório foram realizadas nos tempos de 12, 24 e 48 horas após estímulo inflamatório (HPE), para as seguintes análises: variáveis hematológicas, índices de glicose basal, atividade respiratória dos leucócitos sanguíneos (burst oxidativo), características qualitativas e quantitativas do exsudato, atividade lítica das proteases do soro e atividade de aglutinação bacteriana. Os resultados foram comparados por meio de análise de variância (ANOVA) ao nível de 5% de probabilidade e a diferença entre as médias foi comparada pelo teste de Tukey. Não foram observadas variações hematológicas significativas entre os grupos de tilápias-do-Nilo tratadas e estimuladas e o grupo controle, o mesmo pode ser observados...
The use of probiotics arises as an alternative to the use of antibiotics in fishes, preventing them from becoming sick. The scope of this study was to evaluate the effects of food with added probiotic containing Bacillus amyloliquefaciens in acute inflammation against A.hydrophila in the Nile-tilapia (Oreochromis niloticus). To that end, there were used 936 copies of the Nile-tilapia (30g), evenly distributed in 12 net tanks with 78 fish in each. The fishes were fed for 60 days with food added with the probiotic bacteria in different concentrations, with the following groups being formed; Group 1 (control) fed with basal food without added probiotic: Group 2 (T1) supplemented with 0.75 grams of probiotic / kg food; Group 3 (T2) supplemented with 1.5 grams of the probiotic / kg food; Group 4 (T3) supplemented with 3 grams of probiotic / kg food. For the inflammatory stimulus it was inoculated in the swimming bladder of fish 0.5 ml of saline solution 0.65% in the control group and 0.5 ml x106 of a solution containing inactivated A. hydrophila in the other groups. Blood collections and inflammatory exudate were held in the periods of 12, 24 and 48 hours after the inflammatory stimulus (HPE), for the following analysis: haematological variables, basal glucose levels, respiratory activity of sanguine leukocytes (oxidative burst), qualitative and quantitative characteristics of the exudate, lytic activity of serum proteases and agglutination bacterial activity. The results were compared using analysis of variance (ANOVA) at 5% probability and the difference between the means was compared by Tukey test. There were no significant hematological changes between the groups of Nile-tilapia treated and stimulated and the control group, the same thing can be observed in glycemic indeces and respiratory activity of blood leucocytes. The inflammatory exudate was characterized by the predominant accumulation ...

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Schulz, Denys. "Caracterização parcial do extrato bruto produzido por Bacillus amyloliquefaciens e ensaios preliminares de citotoxicidade." Florianópolis, SC, 2003. http://repositorio.ufsc.br/xmlui/handle/123456789/85760.

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Dissertação (mestrado) - Universidade Federal de Santa Catarina, Centro de Ciências Agrárias. Programa de Pós-Graduação em Ciência dos Alimentos.
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Levando-se em consideração os grandes índices de morbidade causados pela contaminação de alimentos, fica evidente a necessidade de desenvolver novas alternativas de conservação para disponibilizar para a população alimentos de qualidade cada vez melhor e mais seguros, do ponto de vista microbiológico e toxicológico. Frente a essa realidade, esse estudo teve como objetivo otimizar a técnica de extração da substância antimicrobiana e sua caracterização bem como realizar uma avaliação preliminar da citotoxicidade do extrato bruto produzido por Bacillus amyloliquefaciens. O extrato bruto foi obtido de uma cultura de Bacillus amyloliquefaciens em caldo triptona de soja suplementado com extrato de levedura, incubado a 35oC por 24 h, precipitado com sulfato de amônio, ressuspendido em tampão fosfato 0,2 M pH 7,2, dialisado em membrana de 3,5 kDa e esterilizado por filtração em membrana de 0,22 mm. O mesmo procedimento foi repetido substituindo o tampão fosfato por água destilada. Todos os ensaios desse estudo foram realizados com dois pools de extrato bruto, sendo um obtido pela mistura de extratos brutos extraídos em tampão e o outro obtido pela mistura de extratos brutos extraídos em água. O extrato bruto foi submetido aos seguintes ensaios de caracterização: atividade antimicrobiana em ágar triptona de soja suplementado com extrato de levedura e em ágar sangue, perfil de proteínas em gel de poliacrilamida e aminograma. Como microrganismo indicador foi utilizado Listeria monocytogenes a 105 UFC/mL. Foram realizados ensaios preliminares de citotoxicidade como atividade hemolítica em tubos e em placas de ágar sangue e citotoxicidade em células VERO. Nos ensaios de atividade antimicrobiana em ágar triptona de soja suplementado com extrato de levedura o extrato bruto apresentou um halo de inibição máximo de 8,3 mm (profundidade) e 7,5 mm (superfície) numa concentração de 80 mg e 10 mg de proteína, respectivamente; em ágar sangue observou-se um halo de inibição máximo de 4,5 mm (profundidade) e 6,6 mm (superfície) numa concentração de 80 mg e 10 mg de proteína, respectivamente. Para determinação da composição global de aminoácidos em eletroforese PAGE (7,5 a 20%)/SDS (10%) foram constatadas seis bandas com pesos moleculares aparentes de: 14,8; 20; 29; 30; 34 e 56,2 kDa e composto principalmente de aminoácidos hidrofóbicos (não polares), incluindo a prolina (446,77 ppm), leucina (11,03 ppm), valina (6,27 ppm), isoleucina (4,59 ppm), alanina (2,91 ppm) e glicina (1,87 ppm). A quantidade de aminoácidos totais presentes no extrato bruto foi de 526,59 ppm. Em ensaios de citotoxicidade, o extrato bruto apresentou uma concentração hemolítica 50% em tubos na concentração de 11,43 mg de proteína numa suspensão de eritrócitos de carneiro e uma concentração citotóxica 50% na concentração de 32,14 mg de proteína em células VERO; o halo de hemólise máximo em placas de ágar sangue foi de 5,6 mm (profundidade) e 5,7 mm (superfície) numa concentração de 80 mg e 10 mg de proteína, respectivamente. Os resultados obtidos sugerem que o extrato bruto produzido por Bacillus amyloliquefaciens é composto principalmente de proteases e outras enzimas bacteriolíticas.

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Linardi, Valter Roberto. "Melhoramento de Bacillus amyloliquefaciens por transformação e fusão, para produção de a-amilase e proteinas." [s.n.], 1985. http://repositorio.unicamp.br/jspui/handle/REPOSIP/255837.

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Orientador: Yong Kun Park
Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos e Agricola
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Resumo: Utilizando-se duas espécies distintas de Bacillus, B. amyloliquefaciens ATCC 2342 e o B. natto, isolado do produto comercial "Natto", respectivamente, bons produtores de ?-amilase e proteases, conseguiu-se aumentar os níveis de produção destas enzimas, através das técnicas de transformação e fusão de protoplastos. O melhoramento genético por transformação mostrou ser mais eficiente do que o realizado por fusão de protoplastos. Dos vários recombinantes isolados, obtidos por transformação, foram selecionadas as linhagens T-41 e T-39. A primeira, T-41, produziu duas vezes mais ?-amilase e três vezes mais proteases, enquanto que a segunda, T-39, produziu quatro vezes mais proteases, considerando-se o receptor B. amyloliquefaciens. Estudos comparativos demonstraram que a ? - amilase produzida pelo receptor apresentou atividade ótima, em pH 6,0 e temperatura ótima a 65°C. Na temperatura de 60°C a enzima foi estável por 60 minutos. Por outro lado, a enzima produzida pelo T-41 apresentou atividade ótima em pH 6,0 e temperatura ótima a 65°C. A 60°C a atividade amilásica começou a decrescer a partir de 40 minutos, com declínio acentuado a partir de 65°C. A análise do cromatograma evidenciou que as enzimas produzidas por essas linhagens apresentaram a mesma maneira de atuação sobre o amido.O recombinante T-41 mostrou ser mais susceptível a autólise celular do que o B. amyloliquefaciens. A atividade amilásica da linhagem F-52, obtida por fusão de protoplastos, foi o dobro da atividade do parental B. natto, e somente este recombinante desenvolveu a capacidade de crescer a 50°C
Abstract: Two different strains of Bacillus3 were used B. amyloliquefaciens ATCC 2342 and B. natto, isolated from the commercial product "Natto", good producer of ? -amylase and of proteases, respectively. The level of production of these enzymes was increased through technique using transformant DNA and fusion of protoplasts. The genetic improvement by transformation was shown to be more eficient than that obtained by protoplast fusion. Among the various recombinant DNA's isolated and obtained by transformation, the T-41 and T-39 strains were selected. The former produced twice as much a-amylase and three times the quantity of proteases as the B. amyloliquefaciens, while the T-39 strain produced four times as much proteases as B. amyloliquefaciens. Comparative studies demonstrated that the a-amylase produced by the recipient had maximum activity at pH 6,0 and 65°C. At 60°C, the enzyme was stable for 60 minutes. Similarly, the enzyme produced by T-41 showed maximum activity at pH 6,0 and 65°C. At 60°C, the amylase activity began to decrease after 40 minutes, with a sharp decline in activity at 65°C. An analysis of the chromatogram indicated that the enzymes produced by these strains demonstrated the same type of activity on starch. The recombinant T-41 was shown to be more susceptible to cellular autolysis than B. amyloliquefaciens. The amylase activity of the F-52 strain, obtained by protoplast fusion, was twice that of the parent B. natto and only this recombinant developed the capacity to grow at 50°C
Doutorado
Mestre em Ciência de Alimentos

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Kierul, Kinga. "Comprehensive proteomic study of Bacillus amyloliquefaciens strain FZB42 and its response to plant root exudates." Doctoral thesis, Humboldt-Universität zu Berlin, Mathematisch-Naturwissenschaftliche Fakultät I, 2013. http://dx.doi.org/10.18452/16805.

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Bacillus amyloliquefaciens FZB42 ist ein frei lebendes Bakterium, das Pflanzenwurzeln besiedelt und das Pflanzenwachstum durch viele verschiedene Wirkmechanismen anregt. In dieser Arbeit wurden die molekularen Grundlagen dieser positiven Wirkungen, die dieses „Pflanzenwachstum fördernde Rhizobakterium“ (PGPR) auf seine Wirte ausübt, untersucht. Um den gegenseitigen Austausch von B. amyloliquefaciens und seinen Wirtspflanzen zu entschlüsseln, wurden umfangreiche Proteomstudien durchgeführt. Es wurden Referenzkarten der extrazellulären und zytosolischen Proteinfraktionen erstellt. Die größte Anzahl an ausgeschiedenen Proteinen konnte während der stationären Phase beobachtet werden. Die identifizierten extrazellulären Proteine gehören verschiedenen Funktionsklassen an, wobei die prominentesten Klassen am Kohlenhydrat-Abbau und den Transport von Molekülen durch die Zellwand beteiligt sind. Die zytosolischen Extrakte von Kulturen, die in 1C-Medium bzw. Mineralmedium angezogen wurden, und in der zweidimensionalen Gelelektrophorese (2 DE) aufgetrennt wurden, ergaben 461 und 245 verschiedene Protein-Einträge. Die erstellten Referenz-Karten wurden anschließend verwendet, um Proteine und Prozesse, in an der Interaktion mit Pflanzen beteiligt sind, zu identifizieren. Dafür wurden die Bakterien Wurzelexudaten von Mais (Zea mays L.) ausgesetzt. Die Proteine aus zwei Stämmen, denen die globalen Transkriptionsregulatoren (Degu, AbrB) und vier Sigma-Faktoren (SigB, SigM, SigV, und SigX) fehlen, wurden ebenfalls untersucht, um ihre Beteiligung an den bakteriellen Reaktionen auf die Wurzelausscheidungen zu analysieren. Zusammenfassend ist dies die erste Studie, die umfangreiche Proteomdaten von Gram-positiven PGPR präsentiert, wobei gleichzeitig die Veränderung der Expression von extrazellulären und zytoplasmatischen Proteinen, nach Zugabe von Wurzelexudaten, ausgewertet wurde.
Bacillus amyloliquefaciens strain FZB42 is a free-living bacterium that competitively colonizes plant roots and stimulates plant growth by many different modes of action. The molecular basis of singular beneficial effects that this Plant Growth-Promoting Rhizobacteria (PGPR) exert on their hosts have been studied. To decipher the molecular cross-talk of B. amyloliquefaciens and its’ host plants as a whole system, an extensive proteomic approach was performed. Reference maps of the extracellular and cytosolic protein fractions were established. The highest number of secreted proteins was observed during stationary growth phase. Identified extracellular proteins belong to different functional classes, with the most prominent classes involved in carbohydrate degradation and transportation of molecules across the cell wall. Cytosolic extracts obtained from cultures grown in 1C and minimal media subjected to the 2 Dimensional Electrophoresis (2 DE), revealed 461 and 245 different protein entries, respectively. Created reference maps were subsequently used to identify proteins and processes involved in the interaction with plants, prior to exposure of bacteria to maize (Zea mays L.) root exudates. The proteomics of two strains lacking expression of genes coding for global transcriptional regulators (degU, abrB) and four sigma factors (sigB, sigM, sigV, and sigX) were also inves-tigated, in order to analyse their involvement in bacterial responses to root exudates. In summary, this is the first study presenting comprehensive proteomics of Gram-positive PGPR, evaluating at the same time changes in protein expression caused by addition of root exudates at the extracellular and cytosolic level.

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Santos, Tatiana da Silva. "Inoculação via foliar de bactérias diazotróficas em milho cultivado sob diferentes manejos de solo." Ilha Solteira, 2018. http://hdl.handle.net/11449/180597.

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Orientador: Ricardo Antonio Ferreira Rodrigues
Resumo: O manejo eficiente do solo pode provocar mudanças benéficas na qualidade física, química e microbiológica do solo refletindo em alta produtividade de grãos de milho. O nitrogênio (N) é o nutriente mais exigido pela planta de milho. A aquisição de fertilizantes nitrogenados pode ser responsável por 50% dos custos variáveis da lavoura. Dessa forma, a utilização de bactérias diazotróficas é considerado uma fonte alternativa economicamente viável e sustentável para aquisição desse nutriente por meio da fixação biológica de N por esses microrganismos. Em vista do exposto, o objetivo deste trabalho foi verificar o desenvolvimento, componente produtivo e produtividade de grãos de milho sob diferentes manejos de solo e inoculado via foliar por Azospirillum brasilense e/ou Bacillus amyloliquefaciens. O experimento foi conduzido durante o ano agrícola de 2016/17 em um Latossolo Vermelho distroférrico localizado na Fazenda de Ensino, Pesquisa e Extensão (FEPE) – Setor de Produção Vegetal pertencente à Faculdade de Engenharia de Ilha Solteira, município de Selvíria - MS. O delineamento experimental utilizado foi o de blocos casualizados, esquema de faixas. Foram estabelecidos 12 tratamentos com 4 repetições. Os tratamentos foram constituídos por 3 tipos de sistemas de manejo do solo: Sistema convencional (SC), cultivo mínimo (CM) e sistema plantio direto (SPD). Os tratamentos de inoculação via foliar de bactérias diazotróficas consistiram em: Azospirillum brasilense (AB), Bacillus amylol... (Resumo completo, clicar acesso eletrônico abaixo)
Abstract: Efficient soil management can lead to beneficial changes in the physical, chemical and microbiological quality of the soil, reflecting high yields of corn grains. Nitrogen (N) is the nutrient most required by the corn plant. The acquisition of nitrogen fertilizers can account for 50% of the variable costs of the crop. Thus, the use of diazotrophic bacteria is considered an economically viable and sustainable alternative source for the acquisition of this nutrient through the biological fixation of N by these microorganisms. In view of the above, the objective of this research was to verify the development, productive component and productivity of corn grains under different soil management and foliar inoculation by Azospirillum brasilense and / or Bacillus amyloliquefaciens. The experiment was conducted during the agricultural year of 2016/17 in a Distroferric Red Oxisol located at the Teaching, Research and Extension Farm (FEPE) - Vegetable Production Sector belonging to the Faculty of Engineering of Ilha Solteira, Selvíria - MS. The experimental design was a randomized complete block design. Twelve treatments with four replicates were established. The treatments were constituted by 3 types of soil management systems: Tillage (SC), minimum tillage (CM) and notillage (SPD). The treatments of foliar inoculation of diazotrophic bacteria consisted of: Azospirillum brasilense (AB), Bacillus amyloliquefaciens (BA), Azospirillum brasilense + Bacillus amyloliquefaciens (AB + BA) and... (Complete abstract click electronic access below)
Mestre

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Bessler, Cornelius. "Die Alpha-Amylase aus Bacillus amyloliquefaciens : Verbesserung der Alkaliaktivität und Steigerung der spezifischen Aktivität mittels gerichteter Evolution /." [S.l. : s.n.], 2002. http://www.bsz-bw.de/cgi-bin/xvms.cgi?SWB10447199.

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Zecchin, Vivian Jaskiw Szilagyi. "Uso da bactéria promotora do crescimento vegetal, bacillus amyloliquefaciens SUBSP, plantarum FZB-42, tomateiro em cultivo orgânico." reponame:Repositório Institucional da UFPR, 2016. http://hdl.handle.net/1884/44121.

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Orientador : Prof. Dr. Átila Francisco Mógor
Coorientador : Profª. Drª. Lucimeris Ruaro
Tese (doutorado) - Universidade Federal do Paraná, Setor de Ciências Agrárias, Programa de Pós-Graduação em Agronomia. Defesa: Curitiba, 14/06/2016
Inclui referências: f 24-28;31-32;42-45;53-55;68-71;84-88;93-95
Área de concentração: Produção vegetal
Resumo: O tomateiro é uma cultura amplamente difundida, seus frutos podem ser utilizados in natura ou industrializados. Portanto possui relevância econômica e importância social pois seus derivados passam por uma extensa cadeia produtiva que gera inúmeros empregos. Atualmente com uma maior conscientização ambiental e também visando a saúde, o uso de tecnologias biológicas que incrementem a produtividade sem aditivos químicos, são alternativas desejáveis. Neste contexto o uso de bactérias promotoras de crescimento vegetal vem de encontro a estas necessidades. A bactéria Bacillus amyloliquefaciens subsp. plantarum FZB42, que já tem caracterizada algumas habilidades de interesse agronômico, foi o alvo utilizado no estudo da interação planta x bactéria. Sendo assim, os objetivos deste estudo foram: (i) investigar na bactéria características relacionadas à promoção do crescimento vegetal; (ii) verificar alterações morfométricas das plantas de tomateiro inoculadas com FZB42 na germinação de sementes, na produção de mudas e no crescimento das plantas até 60 dias em sistema orgânico em diferentes doses (1,5x109; 6,0x1010; 2,4x1011 bactérias mL-1); (iii) analisar alterações bioquímicas e nutricionais em plantas de tomateiro; (iv) avaliar a produção de tomates mediante inoculação da bactéria nas doses (1,5x109; 6,0x1010). FZB42 apresentou resultados positivos para produção de sideróforos e compostos indólicos com e sem suplementação de triptofano. Sementes inoculadas não tiverem seu percentual de germinação alterado. Na produção de mudas, a dose de 6,0x1010, foi a mais estimuladora do crescimento vegetal, com incrementos na parte aérea das cultivares Cereja 261, Santa Clara I-5300, Santa Cruz Kada Gigante e Serato F1, bem como aumentou os teores de clorofila. As mudas das duas últimas cultivares, na mesma dose, também tiveram o sistema radicular estimulado e os teores de açúcares solúveis e proteínas solúveis aumentados. A dose de 1,5x109 reduziu o crescimento das mudas. Plantas com 60 dias após semeadura, de 'Cereja 261', 'Santa Clara I-5300' e 'Serato F1' apresentaram-se com maior parte aérea e raízes quando inoculadas com 6,0x1010. Plantas de 60 dias submetidas a 1,5x109 tiveram estímulos mais discretos e quando inoculadas com 2,4x1011 os aspectos biométricos foram diminuídos. Nesta etapa o perfil metabólico dos cultivares não seguiu um padrão, pois estavam sujeitos às variação morfométricas devido às diferenças de estadio fenológico. O cultivar Serato F1 foi conduzido à campo em cultivo protegido. Aos 135 dias após plantio (em plena frutificação) apresentou, nas folhas, açúcares solúveis totais e aminoácidos livres totais em maiores quantidades nas plantas inoculadas com as doses de 1,5x109 e 6,0x1010. Já os frutos maduros das plantas inoculadas, também colhidos no mesmo momento, apresentaram mais proteínas solúveis totais e aminoácidos livres totais. Neste mesmo período a análise de macro e micro nutrientes revelou que plantas inoculadas tinham maiores teores de nitrogênio, ferro e manganês. A dose de 6,0x1010 diminuiu os níveis de cobre e aumentou zinco. Quanto aos aspectos produtivos, todos os tratamentos com plantas inoculadas apresentaram cerca de quarto frutos a mais por planta, sendo estes frutos de maior calibre e mais pesados que o controle não inoculado. Isso refletiu em uma produção de cerca de 1 kg a mais por planta, o que considerando 12,820 plantas ha-1, representa um incremento de 11,76 e 13,23 ton ha-1 nas doses de 1,5x109 e 6,0x1010, respectivamente. Portanto, FZB42 na dose de 6,0x1010 foi mais adequado para estímulo ao crescimento vegetal na fase de produção de mudas. Em termos de produtividade ambas as doses 1,5x109 e 6,0x1010 foram eficazes. Sendo assim, FZB42 na dose de 6,0x1010 bactérias mL-1 foi efetivo na promoção de crescimento durante todo o ciclo do tomateiro e com isso demonstrou potencial como inoculante ou bio-fertilizantes proporcionando alterações metabólicas e nutricionais. Palavras-chave: Bactérias promotoras de crescimento vegetal. Inoculante. Bio-fertilizante. Produtividade. Bacillus amyloliquefaciens. Solanum lycopersicum.
Abstract: The tomato is a widespread crop, its fruits can be used as a raw material for processed food. Therefore it has economic relevance and social importance because is the basis of an extensive production chain that generates numerous jobs. Currently greater environmental awareness and also aimed at health, the use of biological technologies that increase productivity without chemical additives, are desirable alternatives. In this context the use of plant growth promoting bacteria comes to attend these needs. The bacteria Bacillus amyloliquefaciens subsp. plantarum FZB42, which already has characterized some desirable agronomically desirable effects, was the aim at this study of plant x bacteria interaction. Thus, the objectives of this study were: (i) investigate the bacteria characteristics related to the promotion of plant growth; (ii) verify morphometric changes of tomato plants inoculated with FZB42 on seed germination, in the production of seedlings, and plant growth to 60 days after sowing in organic system at different doses (1.5x109; 6.0x1010; 2.4x1011 bacteria mL-1); (iii) analyze biochemical and nutritional changes in tomato plants; (iv) evaluate the production of tomatoes by inoculation of bacteria in doses (1.5x109; 1,6x105). FZB42 tested was positive for siderophore production and indolec compounds with and without tryptophan supplementation. Inoculated seeds have not changed their germination percentage. In the production of seedlings, the dose of 1.6x105 was the most stimulating plant growth, with increases in the shoots of 'Cherry 261', 'Santa Clara I-5300', 'Santa Cruz Kada Gigante' and 'Serato F1', as well as increased levels of chlorophyll. The last two cultivars seedlings at the same dose also had their roots stimulated and showed increaes in total soluble sugars and total soluble proteins. The dose of 2.4x1011 reduced seedling growth. Plants 60 days after sowing, the 'Cherry 261,' 'Santa Clara I-5300' and 'Serato F1' were improved their shoots and roots growth when inoculated with 6.0x1010. Plants with 60 days inoculates with 1.5x109 had more discrete stimuli and when inoculated with 2.4x1011 the growth were diminished. At this stage the metabolic profile of cultivars did not follow a pattern because they were subject to the morphometric variation due differences in phenological stages. The cultivar Serato F1 was cultivated in greenhouse. At 135 days after planting (in full fruiting) the leaves presented total soluble sugars and total free amino acids in a larger amount in the plants inoculated with doses of 1.5x109 and 6.0x1010. In addition, the ripe fruits of inoculated plants, harvested at this time, had more total soluble protein and total free amino acids. In the same period, the analysis of macro and micronutrients on leaves showed that inoculated plants had higher levels of nitrogen, iron and manganese. The dose of 6.0x1010 decreased copper and increased zinc levels. All inoculated plants showed the increment of around four fruits per plant, which fruits with greater caliber and heavier than uninoculated control. This reflected in an increasing of about 1 kg at more per plant, considering that 12,820 plants ha-1, representing an increase of 11.76 tons ha-1 and 13.23 tons ha-1 at the doses of 1.5x109 and 6.0x1010 respectively. Therefore, Bacillus amyloliquefaciens FZB42 at 6.0x1010 dose was more efficient for stimulating plant growth in seedling production. In terms of yield, both 1.5x109 and 6.0x1010, doses were effective. Thus, FZB42 at a dose of 6.0x1010 bacteria mL-1 was effective in promoting growth throughout the tomato cycle and it showed potential as inoculant or bio-fertilizer providing nutritional and metabolic changes. Key-words: Plant growth promoting bacteria. Inoculation. Biofertilizers. Productivity. Bacillus amyloliquefaciens. Solanum lycopersicum.

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Van, Staden Anton Du Preez. "In vitro and In vivo characterization of Amyloliquecidin, a novel two-component lantibiotic produced by Bacillus amyloliquefaciens." Thesis, Stellenbosch : Stellenbosch University, 2015. http://hdl.handle.net/10019.1/96656.

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Thesis (PhD)--Stellenbosch University, 2015.
ENGLISH ABSTRACT: Antimicrobial resistance is one of the major problems faced by the medical industry today. The ability of bacteria to rapidly acquire resistance against antibiotics and the over prescription and inappropriate use of antibiotics further exacerbate this crisis. Few new antimicrobials are, however, making it through the drug discovery pipeline. The search and development of novel and effective antimicrobials is therefore of the utmost importance. Lantibiotics are ribosomally synthesized cationic antimicrobial peptides with extensive post-translational modifications. They are active against a wide range of Gram-positive bacteria, including antibiotic-resistant strains. They are characterized by the presence of lanthionine and methyllanthionine rings and have been suggested as alternatives or for use in conjunction with antibiotics against resistant pathogens. Staphylococcus aureus is the most common bacteria isolated from skin and soft tissue infections (SSTIs). Strains of S. aureus have emerged with resistance against antibiotics with the most common being methicillin-resistant S. aureus (MRSA). Several lantibiotics are active against MRSA in vivo and have even shown superior activity to traditional antibiotics. Lantibiotics therefore show much promise for the treatment of SSTIs caused by resistant- and non-resistant S. aureus. In this study the bacterially diverse soil of the Fynbos in the Western Cape was screened for novel antimicrobials. Two antimicrobial producing Bacillus strains were isolated, Bacillus clausii AD1 and Bacillus amyloliquefaciens AD2. Both of these strains produce lantibiotics with B. clausii AD1 producing a known lantibiotic, clausin. B. amyloliquefaciens AD2 produces a novel two-component lantibiotic which was designated amyloliquecidin. The lantibiotic operon of amyloliquecidin was sequenced and annotated. All the genes required for successful production of amyloliquecidin are present in the operon. Amyloliquecidin was characterized in vitro and along with clausin is active against clinical strains of S. aureus (including MRSA), Enterococcus spp., Listeria spp. and beta-haemolytic streptococci. Amyloliquecidin has remarkable stability at physiological pH compared to nisin and clausin. A comparative in vivo murine infection model was used to evaluate the effectiveness of amyloliquecidin, nisin, clausin and Bactroban (commercial S. aureus topical treatment) in treating wound infections caused by S. aureus. All the lantibiotics proved to be just as effective as the Bactroban treatment. Furthermore, the tested lantibiotics did not have a negative influence on the wound closure rates of infected and non-infected wounds. Bactroban had a negative effect on wound healing compared to the lantibiotics. To our knowledge amyloliquecidin is the third two-component lantibiotic isolated from Bacillus. This study represents the first to test the effectiveness of amyloliquecidin in vivo and is one of a handful to test lantibiotics as topical treatments.
AFRIKAANSE OPSOMMING: Antimikrobiese weerstandbiedende bakterieë is op die oomblik een van die grootste probleme in die mediese veld. Die antibiotika krisis word vererg deur die vermoë van bakterieë om vinnig weerstand op te bou teen antibiotika, asook die alledaagse misbruik van antibiotika. Daar is ook ŉ tekort in die hoeveelheid antibiotika wat na die finale fases van ontwikkeling gaan. Om die oorhand teen antibiotika-weerstandige bakterieë te kry is dit van uiterste belang dat meer effektiewe antibiotika ontdek word. Lantibiotika is kationiese antimikrobiese peptiede wat deur die ribosoom gesintetiseer word en bevat ŉ verskeidenheid van modifikasies wat na translasie ingebou word. Hulle word gekarakteriseer deur lanthionien en metiellanthionien ringe. Lantibiotika is aktief teen ŉ verskeidenheid Gram-positiewe bakterieë en kan in kombinasie met antibiotika, of as alternatief gebruik word. Staphylococcus aureus is die mees algemene bakterium wat geassosieer word met vel en sagte weefsel infeksies (VSWIs). Staphylococcus aureus met weerstand teen antibiotika is ook al geïsoleer, die mees algemene weerstandige ras is methisillien-weerstandige S. aureus (MWSA). Lantibiotika is wel aktief teen MWSA in vitro en in vivo, met van hulle wat tot beter aktiwiteit as die voorgeskrewe antibiotika het. Lantibiotika kan dus gebruik word as behandeling vir VSWIs wat veroorsaak word deur weerstandige S. aureus, asook teen nie-weerstandige rasse. In hierdie studie was die bakteriese diverse grond van die Fynbos in die Wes-kaap ondersoek vir bakterieë wat antimikrobiese middels produseer. Twee Bacillus rasse, Bacillus clausii AD1 en Bacillus amyloliquefaciens AD2, wat antimikrobiese middels produseer, is geïsoleer. Bacillus clausii AD1 produseer ŉ bekende lantibiotikum, naamlik clausin. Bacillus amyloliquefaciens AD2 produseer ŉ nuwe twee-komponent lantibiotikum, amyloliquecidin. Die lantibiotikum operon wat verantwoordelik is vir die produksie van amyloliquecidin is geïdentifiseer en geannoteer. Die operon bevat al die gene benodig vir die biosintese van amyloliquecidin. Amyloliquecidin is in vitro gekarakteriseer en het aktiwiteit teen ŉ verskeidenheid Gram-positiewe bakterieë. Amyloliquecidin en clausin is aktief teen S. aureus (insluitend MWSA), Enterococcus spp., Listeria spp. en beta-hemolitiese streptococci wat vanaf infeksies geïsoleer is. Amyloliquecidin is baie stabiel by filologiese pH en aansienlik meer stabiel as nisin en clausin. Die effektiwiteit van nisin, clausin en amyloliquecidin in die behandeling van muis vel infeksies veroorsaak deur S. aureus was vergelyk met die kommersiële behandeling Bactroban. Al drie lantibiotika het die verspreiding van S. aureus met die selfde effektiwiteit as Bactroban belemmer. Geen van die lantibiotika het ŉ negatiewe effek op wond genesing nie. Bactroban, inteendeel, belemmer wond genesing. So ver ons weet is amyloliquecidin die derde twee-komponent lantibiotikum wat uit Bacillus geïsoleer is. Die studie is ook die eerste om die effektiwiteit van amyloliquecidin in vivo te rapporteer, asook ook een van die min studies wat kyk na lantibiotika as behandeling vir topikale infeksies.

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Massi, Juliana Barion. "Farelo de soja e casca de arroz para a produção de lipopetideos por Bacillus amyloliquefaciens MO.04b." Universidade Estadual de Londrina. Centro de Ciências Exatas. Programa de Pós-Graduação em Biotecnologia, 2014. http://www.bibliotecadigital.uel.br/document/?code=vtls000200440.

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Resíduos agroindustriais como bagaços, farelos, cascas entre outros podem ser matéria prima e fonte de energia para a obtenção de diversos produtos com alto valor agregado. Entre eles os biossurfactantes, metabólitos microbianos de grande interesse industrial por suas inúmeras aplicações e pelas vantagens ambientais de baixa toxicidade e alta biodegradabilidade, mas com utilização limitada pelo alto custo de produção. O objetivo deste estudo foi avaliar a produção de lipopetídeos, uma classe de biossurfactantes, por fermentação em estado sólido (FES), utilizando Bacillus amyloliquefaciens MO.04b. As fermentações foram desenvolvidas a partir de misturas dos substratos em diferentes proporções, com 80 % de umidade. Os frascos inoculados (200 μL de suspensão de células) foram incubados em BOD a temperaturas de 28 a 37 ± 2 °C por até 48 horas. A interrupção dos cultivos foi pela adição de água, a mistura homogeneizada (180 rpm por 30 min.) e centrifugada (10.000 rpm por 20 min, 4 °C). Os lipopeptídeos foram precipitados no sobrenadante obtido pela adição de HCl 6M, (pH 2 a 4 °C); seguido de nova centrifugação o precipitado então foi ressuspenso em água, neutralizado (NaOH 0,5 M, pH 7,0) e liofilizado. Soluções do liofilizado foram preparadas para as determinações: Indice de Emulsificação (IE24); Tensão Superficial (TS); Infravermelho com Transformada de Fourrier (IV-FT); Espectroscopia de Ressonância Magnética Nuclear (RMN de 1H e 13C) e Espectrometria de Massa. Os substratos selecionados para a produção foram o farelo de soja (47 % de proteínas) 4 g e a casca de arroz utilizada comobulking agent 3,75 g. A avaliação individual dos parâmetros de cultivo com os seguintes resultados, umidade 80 %; solução umedecedora MSM-1; fonte de nitrogênio extrato de levedura 1,75 g; fonte de carbono glicerol 1,50 g; pH inicial 6. O farelo de soja, extrato de levedura e glicerol, foram os pontos centrais de um planejamento fatorial Box-ehnken 33, onde a condição ótima de produção de lipopeptídeos foi 44,072 mg/g.s.s, em farelo de soja (3,25 g), glicerol (1,50 g), extrato de levedura (1,75 g) a 37 ± 2 °C , e 36 h de incubação. As análises de IV-FT apresentaram espectro semelhante ao da surfactina padrão, sugerindo a presença deste biossurfactante, que é produzido pelo B. amyloliquefaciens MO.04b em fermentação submersa. Quanto as determinações de RMN e espectrometria de massa não foram conclusivas, provavelmente pela complexidade da amostra e a necessidade de purificação da mesma.
Agroindustrial residues as bagasse, bran, hulls and other raw materials, can be an energy source for obtaining various products with high added value. Among them biosurfactants microbial metabolites, of great industrial interest for numerous applications, and the environmental advantages of low toxicity and high biodegradability, but limited by the high cost of production. The goal of this study was to evaluate the production of lipopetídeos, a class of biosurfactants, by solid state fermentation (SSF) using Bacillus amyloliquefaciens MO.04b. Fermentations were developed from mixtures of substrates in different proportions, with 80 % humidity. The inoculated flasks (200 μl cell suspension) were incubated in a temperature chamber at 28-37 ± 2 ° C for 48 hours. The interruption of the cultures was the addition of water, the homogenized mixture (180 rpm for 30 min.) centrifuged (10,000 rpm for 20 min, 4 ° C). The lipopeptides were precipitated in the obtained supernatant by adding 6M HCl ( pH 2 at 4 ° C); followed by further centrifugation the precipitate was then resuspended in water, neutralized (0.5 M NaOH , pH 7.0) and lyophilized. Solutions were prepared for determination: Emulsification Index (IE24) ; Surface Tension (TS) ; Fourier Transform Infrared (FT-IR); Nuclear Magnetic Resonance Spectroscopy (1H NMR and 13C NMR), and mass spectrometry. Selected to produce substrates were soybean meal (47 % protein), 4 g, rice husk bulking agent 3.75 g. The evaluation of individual cultivation parameters with the following results, 80 % humidity; solution MSM- 1; nitrogen sources yeast extract 1.75 g; carbon source glycerol 1.50 g; initial pH 6. Soybean meal , yeast extract and glycerol , were the focal points of a factorial design Box Behnken 33 , where the optimum condition for the production of lipopeptides was 44.072 mg/gds in soybean meal (3.25 g), glycerol (1.50 g) , yeast extract (1.75 g) at 37 ± 2 ° C and 36 h incubation . The FT-IR analysis showed a similar pattern to that of surfactin spectrum indicating the presence of this biosurfactant, which is produced by B. amyloliquefaciens MO.04b in submerged fermentation. Regarding the determination of NMR and mass spectrometry were not conclusive, probably due to the complexity of the sample and the need for purification thereof.

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Chen, Xiaohua. "Whole genome analysis of the plant growth-promoting Rhizobacteria Bacilllus amyloliquefaciens FZB42 with focus on its secondary metabolites." Doctoral thesis, Humboldt-Universität zu Berlin, Mathematisch-Naturwissenschaftliche Fakultät I, 2010. http://dx.doi.org/10.18452/16095.

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Bacillus amyloliquefaciens FZB42 besitzt einen beeindruckenden Effekt zur Verbesserung des Pflanzenwachstums. Um die Mechanismen, vor allem auf molekularer Ebene, zu verstehen, wurde das komplette Genom von FZB42 in dieser Arbeit sequenziert. Abwesenheit von der weit verbreiteten Phagen-verwandten Genen im Genom von B. subtilis 168, der in enger Verwandtschaft zum FZB42 steht, ist ein besonderes Merkmal. Dagegen enthält das Genom von FZB42 viele DNA-Inseln, in denen unikale Gene in FZB42 als Cluster gefunden wurden. Viele Gene, die möglicherweise zur Pflanzenwachstumsförderung beitragen, wurden in dieser Arbeit identifiziert. B. amyloliquefaciens FZB42 ist natürlich kompetent. Das kompetente Stadium in FZB42 kommt früher als in B. subtilis 168, nämlich während der späten exponentiellen Wachstumsphase. Das FZB42-Genom enthält den kompletten Satz von Genen, die für die Entwicklung der genetischen Kompetenz nötig sind. Ausgenommen von Gene für Quorum-Sensing-System ist die Mehrzahl der Kompetenz-Gene von FZB42 sehr ähnlich zu denen in B. subtilis 168. Das FZB42 Genom birgt ein enormes Potential zur Produktion von sekundären Metaboliten. Genetische Manipulationen wurden durchgeführt, um die Funktionen der trans-AT Domänen und der Modifikationsdomänen in den PKS-Gen-Clustern zu erklären. Mit Ausnahme von fünf Gen-Clustern in B. subtilis 168 (Surfactin, Fengycin, Bacillibactin, Bacillaene und Bacilysin), sind Bacillomycin D, Difficidin, Macrolactin und ein hypothetisches Tripeptid einzigartig im Genom der FZB42. FZB42 kann kein bekanntes ribosomal synthetisiertes Bacteriocin produzieren kann. Gleichzeitig beinhaltet sein Genom ein Gen-Cluster, das wahrscheinlich für die Produktion eines neuartigen Bacteriocins verantwortlich ist. Die eindrucksvolle genetische Kapazität zur Herstellung von antagonistischen sekundären Metaboliten ermöglicht es FZB42, nicht nur erfolgreich neben konkurrierenden Organismen innerhalb seiner natürlichen Umgebung zu überleben, sondern auch Pflanzen gegen pathogene Bakterien und Pilze zu schützen.
Bacillus amyloliquefaciens FZB42 has an impressive effect to improve plant growth. In order to understand the mechanisms, especially at the molecular biological level, the whole genome of FZB42 was sequenced in this work. The absence of extended phage insertions which are typical for the closely related B. subtilis 168 genome is a particular feature. On the other hand, several DNA islands where unique genes in FZB42 were found clustered. Many candidate genes that may contribute to the plant growth promotion were identified in this works. B. amyloliquefaciens FZB42 is naturally competent. FZB42 exhibited its maximal competence earlier than B. subtilis, during late exponential growth. Not surprisingly, the FZB42 genome harbors the complete set of genes necessary for development of genetic competence. The majority of competence genes are highly homologous to their counterparts in B. subtilis 168, excluded from genes for the quorum-sensing system. The FZB42 genome harbors enormous potential for producing secondary metabolites. Genetic manipulation was carried out to investigate the trans-AT domains and some modification domains in the pks gene clusters. With the exception of five gene clusters in B. subtilis 168 (Surfactin, Fengycin, Bacillibactin, Bacillaene and Bacilysin), Bacillomycin D, Difficidin, Macrolactin and a hypothetical tripeptide are unique in the genome of the FZB42. A remarkable feature of the FZB42 genome is that it does not produce any known ribosomally synthesized bacteriocin, whereas a gene cluster probably responsible for production of a new bacteriocin was identified in this work. The impressive genetic capacity to produce antagonistic acting secondary metabolites not only enables FZB42 to cope successfully with competing organisms within its natural environment, but also to protect plants from pathogenic bacteria and fungi.

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Arasaki, Karine Marie. "Efeito da atividade antimicrobiana de substância produzida por Bacillus amyloliquefaciens no controle da microbiota do mexilhão Perna perna (Linnaeus, 1758)." Florianópolis, SC, 2002. http://repositorio.ufsc.br/xmlui/handle/123456789/84410.

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Dissertação (mestrado) - Universidade Federal de Santa Catarina, Centro Tecnológico. Programa de Pós-Graduação em Ciência dos Alimentos.
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Neste trabalho, avaliou-se a atividade antimicrobiana de substância produzida por Bacillus amyloliquefaciens sobre a microbiota do mexilhão Perna perna cultivado na praia do Sambaqui, na cidade de Florianópolis. Para a realização dos testes de inibição realizaram-se 3 coletas nas quais as amostras de mexilhão foram submetidas inicialmente a Contagem Padrão em Placas para microrganismos psicrotróficos. Para a obtenção do extrato, as culturas estoques de B. amyloliquefaciens foram centrifugadas e saturadas. Após nova centrifugação, o precipitado foi ressuspendido em tampão fosfato, dialisado e esterilizado por filtração. As bactérias isoladas do mexilhão foram cultivadas sobre placas de TSA onde foram feitos 2 poços de 5mm de diâmetro nos quais adicionou-se 80mL da solução inibidora. Foi observada a formação ou não de halos de inibição ao redor dos poços. Das 39 colônias isoladas, 9 apresentaram halos de inibição e foram identificadas como Bacillus sp, Aerococcus viridians, Acinetobacter spp., Corynebacterium spp e Staphylococcus epidermidis. Foi possível verificar que a substância produzida por B. amyloliquefaciens teve uma discreta ação sobre a microbiota do mexilhão Perna pena quando aplicada diretamente na carne do mesmo, apresentando um comportamento bacteriostático. Os gêneros Bacillus sp, Aerococcus viridians, Acinetobacter spp., Corynebacterium vitarumen, apresentaram mudança da cor do indicador do meio Niven modificado, sugerindo a formação de compostos alcalinos.

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Pereira, Murilo Anderson. "Qualidade microbiológica de ostras Crassostrea gigas e estudo da ação sinérgica da substância antimicrobiana produzida por Bacillus amyloliquefaciens." Florianópolis, SC, 2004. http://repositorio.ufsc.br/xmlui/handle/123456789/87812.

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Dissertação (mestrado) - Universidade Federal de Santa Catarina, Centro de Ciências Agrárias. Programa de Pós-Graduação em Ciência dos Alimentos.
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O elevado índice de morbidade em função da contaminação dos alimentos tem aumentado a necessidade do uso de metodologias alternativas para conservação dos mesmos. Nesse sentido, essa pesquisa teve como objetivo: i) descrever uma revisão sobre a ação de antimicrobianos naturais produzidos por microrganismos para conservação de alimentos; ii) avaliar a qualidade microbiológica de ostra Crassostrea gigas produzida e comercializada em diferentes áreas da região litorânea de Florianópolis # SC; iii) Estudar a atividade antimicrobiana do extrato bruto produzido por Bacillus amyloliquefaciens potencializado pela ação do pH e da concentração de sal em estudos simulados in vitro, com intuito de utilizá-la como inibidor de Listeria monocytogenes em ostras. Com a revisão concluiu-se que as substâncias antimicrobianas, em geral, são uma das opções em um mosaico de possíveis mecanismos para controlar bactérias patogênicas e deteriorantes em alimentos. Tudo indica que conservadores naturais, particularmente em adição ou combinação sinergística com outros fatores e técnicas, têm um papel importante na industria de alimentos, garantindo segurança ao consumidor. Para verificar a qualidade microbiológica das ostras, durante o ano de 2003, 90 (noventa) amostras de ostras da espécie Crassostrea gigas foram analisadas, sendo que 45 delas foram coletadas junto a estabelecimentos comerciais destinados à venda de frutos do mar e 45 amostras diretamente no local de cultivo. De acordo com os resultados, Víbrio cholerae, Víbrio parahaemolyticus e Salmonella sp. não foram encontrados em nenhuma das amostras. Apenas uma amostra apresentou 80 UFC/g de estafilococos coagulase positiva, as demais amostras apresentaram <10 UFC/g. Com o resultado das análises de coliformes a 35ºC e coliformes a 45 ºC, evidencia-se contaminação tanto no local de cultivo quanto no local de venda, nas três áreas analisadas. Escherichia coli foi encontrada em 4 amostras analisadas do local de cultivo, demonstrando uma ocorrência de 9 %. Já nos estabelecimentos comerciais, foi encontrada uma ocorrência de 35,5 % (16/45). A ação do extrato bruto produzido por Bacillus amyloliquefaciens em diferentes concentrações de proteína (0,0; 5,0; 40; 80 µg/mL) sobre o inóculo de 106 UFC.mL-1 de Listeria monocytogenes, foi verificada em caldo TSB-YE previamente ajustado para pH 4,0; 6,1; 8,0; e concentrações de NaCl para 0; 2,5; 4,5 % p/v, resultando em uma análise fatorial completa de 36 curvas de comportamento microbiano. Os ensaios foram realizados à temperatura de 25 ºC em estufa DBO, por até 50 horas. Os resultados obtidos indicam que ocorreu uma ação sinérgica entre pH, concentração de sal e extrato bruto, demonstrando uma potencialização da ação antimicrobiana. A melhor ação foi observada quando se utilizou 80 µg/mL de extrato bruto da substância antimicrobiana, pH 4,0 e 4,5 % de concentração de sal, onde ocorreu uma redução de aproximadamente cinco ciclos logarítmicos da cultura de Listeria monocytogenes.

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Al-Ali, Ameen Ghazi Shimal. "Characterization of Bacillus amyloliquefaciens FZB42 and Bacillus subtilis BBG131 properties responsible for their ability to colonize tomato rhizosphere." Thesis, Lille 1, 2016. http://www.theses.fr/2016LIL10020/document.

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Les bactéries qui promeuvent la croissance de plantes (PGPR) font une partie indispensable du biote dans la rhizosphère. Plusieurs espèces de Bacillus appartiennent aux PGPR et ont la capacité de produire des peptides non ribosomiques différents, tels que les lipopeptides cycliques. La surfactine, fengycine et iturine sont des lipopeptides cycliques produits par B. subtilis and B. amyloliquefaciens. Dans cette étude, nous avons mis en avant l’étude de deux facteurs. Premièrement, la croissance bactérienne et la production de surfactine dans la rhizosphère pendant 21 jours. On outre, les effets des exsudats racinaires et des certaines sources de carbone. Finalement, le rôle de la formation de biofilm a été élaboré par la construction d’un mutant qui a perdu la capacité de la formation de biofilm. Deux souches ont été choisies pour cette étude : B. amyloliquefaciens FZB42 est une souche sauvage et B. subtilis BBG131 qui est un micro-organisme génétiquement modifié. La biomasse de B. amyloliquefaciens FZB42 était 25 fois plus élevée qu’avec B. subtilis BBG131, tandis que la production de surfactine était 5 fois plus faible que celle de B. subtilis BBG131. Les deux souches ont la capacité de croitre dans des exsudats racinaires et produire de la surfactine. B. amyloliquefaciens FZB42 a montré la capacité intense un biofilm alors que B. subtilis BBG131 a montré le contraire. Un mutant de B. amyloliquefaciens FZB42 EPS- a été comparé à d’autres souches. Un comportement similaire de B. amyloliquefaciens FZB42 EPS- et B. subtilis BBG131 a été observé dans la formation de biofilm qui se reflète dans leur colonisation dans la rhizosphère
Plant growth promoting bacteria (PGPR) are an indispensable part of rhizosphere biota. Many of Bacillus species belong to PGPR and have the ability to produce different non-ribosomal peptides such as cyclic lipopeptides. Surfactin, Fengycin and Iturin are cyclic lipopeptides produced by B. subtilis, and B. amyloliquefaciens. In this work, we shed light to study these factors. Firstly, bacterial growth and surfactin production in the rhizosphere during 21 days. Additionally, the effects of root exudates and certain carbon sources. Finally, the role of biofilm formation was elaborated by the construction of a mutant that lost the ability to form a biofilm. Two strains were chosen: Bacillus amyloliquefaciens FZB42 which is a natural wild-type strain and Bacillus subtilis BBG131 which is a genetically engineered microorganism. The biomass of B. amyloliquefaciens FZB42 was 25 times higher than with B. subtilis BBG131, whereas surfactin production was 5 times less than this produced by B. subtilis BBG131. The two strains have the ability to grow in the root exudates and produce surfactin. B. amyloliquefaciens FZB42 and B. subtilis BBG131. B. amyloliquefaciens FZB42 showed intense ability to form a biofilm whereas B. subtilis BBG131 showed contrarily. A mutant of B. amyloliquefaciens FZB42 EPS- was compared to other strains. A similar behavior of B. amyloliquefaciens FZB42 EPS- and B. subtilis BBG131 was observed in the biofilm formation which is reflected to their colonisation in the rhizosphere

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Zühlke, Marie-Katherin [Verfasser]. "Charakterisierung der bakteriellen Transformation von Bisphenolen als östrogenen Umweltschadstoffen durch Bacillus amyloliquefaciens und Cupriavidus basilensis / Marie-Katherin Zühlke." Greifswald : Universitätsbibliothek Greifswald, 2017. http://d-nb.info/1135191220/34.

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Fan, Ben. "Plant colonization by GFP-labeled Bacillus amyloliquefaciens FZB42 and transcriptomic profiling of its response to plant root exudates." Doctoral thesis, Humboldt-Universität zu Berlin, Mathematisch-Naturwissenschaftliche Fakultät I, 2011. http://dx.doi.org/10.18452/16267.

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In dieser Arbeit wurden zunächst die Kolonisationen von drei verschiedenen Pflanzengattungen durch den GFP-markierten Bacillus amyloliquefaciens FZB42 mittels confocaler Lasermikroskopie und Elektronmikroskopie verfolgt. Hier konnte gezeigt werden, dass FZB42 alle ausgewählten Pflanzen besiedeln konnte. Bei Arabidopsis- und Maiskeimlingen wurden die Wurzelhaare und Verbindungen, an denen laterale Wurzeln entstehen, durch FZB42 bevorzugt besiedelt. Weiterhin wurden bei Arabidopsis die Spitzen der Primärwurzeln, und bei Mais die Wurzelkerben bevorzugt besiedelt. Bei Lemna wurden FZB42 Zellansammlungen entlang der Furchen, die zwischen den Epidermiszellen der Wurzel liegen, sowie den intrazellulären Hohlräumen an der Blattunterfläche gefunden. Anschließend wurden die Transkriptome von FZB42, der mit Maiswurzelexudat angezogen wurde, mittels Microarray analysiert. Insgesamt wurden 302 Gene, die 8,2 % des Transkriptoms ausmachen, signifikant durch das Wurzelexudat beeinflusst, wobei die Mehrzahl (260 Gene) hochreguliert wurde. Die induzierten Gene, dessen Funktion bereits bekannt ist, sind hauptsächlich an dem Nährstoffwechsel, Chemotaxis und Beweglichkeit, sowie an der Produktion von Antibiotika beteiligt. Auch wurden die Trankriptome von sieben FZB42-Muatnten durch Microarray analysiert. Diese hatten jeweils eine Deletionen in fünf Sigmafaktor-Genen (sigB, sigD, sigM, sigV,and sigX) und zwei globalen Transkriptionsregulator-Genen (degU und abrB). Die Expression vieler Genen wird durch diese Genprodukte beeinflusst. Mögliche Mechanismen, wie diese Faktoren die bakterielle Reaktion auf Wurzelexsudaten beeinflüssen, wurden vorgeschlagen. Schließlich wurden Northernblott-Untersuchungen an möglichen sRNA-Kandidaten durchgeführt, dessen Expression signifikant durch Wurzelexudate beeinflusst wurde. Dabei konnten 6 von 20 vermeintlichen sRNA-Kandidaten betätigt werden. Dies weist auf eine noch unbekannte Rolle der sRNAs bei der Pflanzen-Mikroben-Wechselwirkung.
In this work colonization of three different plants genera, maize, Arabidopsis, and Lemna, by GFP-labeled Bacillus amyloliquefaciens FZB42 in a gnotobiotic system was firtly studied using confocal laser scanning microscopy and electron microscopy. It was shown that FZB42 is able to colonize all these three plants with a specific pattern. Root hairs and the junctions where lateral roots occurred were a preferred area of FZB42 on both maize and Arabidopsis seedlings. On Arabidopsis, tips of primary roots were another favored site of FZB42; while, on maize, the concavities in root surfaces were preferred. FZB42 cells were also able to colonize Lemna, preferably accumulating along the grooves between epidermis cells on roots and the concaved intercellular space on fronds. Secondly, microarray experiments were performed concerning the transcriptomic response of FZB42 to maize root exudates. A total of 302 genes representing 8.2% of FZB42 transcriptome were significantly altered in transcription by the presence of root exudates, the majority of them (260) were up-regulated in expression. The induced genes with known function were mainly involved in nutrition utilization, chemotaxis and motility, and antibiotic production. The transcriptome of seven FZB42 mutants, defective in five sigma factor genes (sigB, sigD, sigM, sigV, and sigX) and two global transcriptional regulator genes (degU and abrB), were also investigated through microarray experiments. A vast number of genes were indentified to be controlled by the protein factors respectively. Possible mechanisms were proposed of how these protein factors are involved in the response to root exudates. Finally, by northern blot existence of six out of 20 small RNA (sRNA) candidates was identified, which were significantly altered in expression by root exudates. This suggests that sRNA may play a hitherto unrecognized role in plant-microbe interaction.

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Boukhris, Ines. "Caractérisation, clonage, expression et étude de la régulation de gènes phytases de Streptomyces et Bacillus." Thesis, Université Paris-Saclay (ComUE), 2015. http://www.theses.fr/2015SACLS274.

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Les phytases hydrolysent les phytates représentant la forme majeure de stockage du P dans les céréales. Ces phytates sont aussi des facteurs anti-nutritionnels qui chélatent les cations réduisant leur absorption. Dans le premier volet de cette thèse, une nouvelle souche bactérienne produisant une phytase extracellulaire a été isolée et identifiée comme Bacillus amyloliquefaciens US573. L’enzyme «PHY US573» a été purifiée et caractérisée en comparaison avec deux phytases commerciales Ronozyme PL et Natuphos. PHY US573 se distingue par sa forte thermostabilité en présence de calcium. En outre, PHY US573 se caractérise aussi par une tolérance remarquable aux sels comme le NaCl et LiCl. L’ensemble de ces propriétés montre que PHY US573 pourrait être une candidate intéressante pour des applications en alimentation animale ou en agriculture pour améliorer la biodisponibilité du P-phytique pour les plantes. Dans le deuxième volet, la souche Streptomyces sp. US42 produisant une activité phytase extracellulaire a été sélectionnée. L’enzyme «PHY US42» a été purifiée et caractérisée. PHY US42 est calcium dépendante également une grande stabilité en présence de sels biliaires et des protéases digestives. La modélisation moléculaire de PHY US42 indique qu'elle appartient au groupe des β-propeller phytases qui sont généralement calcium-dépendantes. Vu ses propriétés biochimiques intéressantes, PHY US42 constitue une bonne candidate comme additif dans les aliments pour animaux monogastriques en combinaison avec une histidine acide phytase. Enfin dans un troisième volet, nous nous sommes intéressés à l’étude de la régulation de l'expression du gène phytase de S. coelicolor M145 (sco7697) chez S. coelicolor M145, S. lividans TK24 ainsi que chez ses deux mutants ppk et phoP. Ainsi, en plus des boites pho localisées en amont de la région promotrice -35 siège de la régulation positive PhoP-dépendante, nous avons révélé pour la première fois que la RD localisée en aval de la région promotrice -10 est le siège d’une forte régulation négative par un répresseur inconnu. Ce dernier empêcherait l’activation PhoP-dépendante de l’expression du gène phytase
Phytases hydrolyse phytate representing the major storage form of P in cereal. phytates are also anti-nutritional factors that chelate cations such as Ca²⁺, Mg²⁺, Fe²⁺, Z²⁺ reducing their absorption. The low bioavailability of phytic phosphorus in monogastric animals require their food supplementation with Pi to meet the needs of the animal in P. This creates an extra cost and increases the environmental pollution by the manure excretion highly charged phosphate. In the first part of this thesis, from soil samples taken near hot hydrothermal waters of the region Elhamma in southern Tunisian, a new bacterial strain producing extracellular phytase was isolated and identified as Bacillus amyloliquefaciens US573. The enzyme referred "PHY US573" was purified and characterized in comparison with two commercial acid histidine phytases Ronozyme PL and Natuphos. PHY US573 is calcium dependent and has an optimum activity at pH 7.5 (5 for Ronozyme and 5.5 for Natuphos) and 70°C (55°C for Ronozyme and Natuphos). PHY US573 is distinguished by its high thermostability, in fact, it keeps 93% of its activity after incubation for 10 min at 75°C in the presence of calcium while Ronozyme and Natuphos keep only 45% and 53% of their activity, respectively. This enzyme is specific for phytic acid and also has a very good stability at pH 3 to 9 and a perfect stability in presence of bile salts. In addition, PHY US573 is also characterized by a remarkable salt tolerance because it retains 80 to 95% of its activity in the presence of 20 g/l of NaCl and LiCl, respectively. All these properties shows that PHY US573 could be an interesting candidate for applications in feed industry alone or in combination with an histidine acid phytase. In a second part of this thesis, from the Streptomyces collection of LMB-CBS, a strain producing extracellular phytase activity was selected and identified as Streptomyces sp. US42. The enzyme "PHY US42" was purified and characterized. PHY US42 has a calcium-dependent activity (such as Bacillus phytases), optimally active at pH 7 and 65°C. PHY US42 is perfectly stable at pHs ranging from 5 to 10 and its thermal stability is greatly increased in the presence of calcium. Indeed, PHY US42 maintains 80% of its activity after 10 min of incubation at 75 °C in the presence of calcium. PHY US42 has also a high stability in the presence of bile salts and digestive proteases. Molecular modeling of PHY US42 indicates that it belongs to the β-propeller phytase group which are usually calcium-dependent. Given its interesting biochemical properties, PHY US42 which would operate mainly in the intestine, is a good candidate for use as an additive in agastriques fish food or in combination with an histidine acid phytase in feed industry. Finally in a third part, we are interested in studying the regulation of the expression of the phytase gene of S. coelicolor M145 (sco7697) in S.coelicolor M145, S.lividans TK24 and among its two mutants ppk and phoP. To do this, we merged the wild promoter regions (phyWT) or mutated (phym1, phym2, phym1+2) of sco7697 gene with the GUS reporter gene encoding ß-glucuronidase activity. Thus as expected, we demonstrated that the deletion of the PHO box located upstream of the -35 reduces the level of induction of sco7697 in conditions of Pi limitation. Moreover, we have revealed for the first time that the alteration of RD located downstream of -10 correlates with a dramatic increase of GUS expression when PhoP is present. Our results demonstrate that this RD is the seat of a strong negative regulation by an unknown repressor. This would prevent the PhoP-dependent activation of expression of the phytase gene

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Gotor, Vila Amparo María. "New advances in the control of brown rot in stone fruit using the biocontrol agent Bacillus amyloliquefaciens CPA-8." Doctoral thesis, Universitat de Lleida, 2017. http://hdl.handle.net/10803/405888.

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La podridura marró causada pel fong Monilinia spp. és responsable d'importants pèrdues en la postcollita de la fruita de pinyol. Entre les estratègies de control respectuoses amb el medi ambient, cal destacar l'ús d'agents de biocontrol (ABCs). Aquesta tesi té com a objectiu completar el desenvolupament de l'ABC B. amyloliquefaciens CPA-8 per així obtenir un producte eficaç que proporcioni una estratègia comercialment viable. Els principals resultats es classifiquen en (i) caracterització de CPA-8 (disseny de marcadors moleculars, respostes ecofisiològiques, sensibilitat a antibiòtics, producció d’enterotoxines); (ii) desenvolupament de productes basats en CPA-8 (millora del medi de creixement, selecció de la tecnologia de formulació, optimització de l'assecat per llit fluïditzat-atomització); i (iii) definició de l'estratègia de control de CPA-8 (activitat en diferents hostes, definició de llindars tècnics i eficàcia en assajos de camp). La integració dels productes basats en CPA-8 en els sistemes de cultiu habituals contribueix en el maneig de les malalties de postcollita en fruita d'os en el marc d'una agricultura sostenible i / o ecològica.\\\\
La podredumbre marrón causada por el hongo Monilinia spp. es responsable de importantes pérdidas en la poscosecha de la fruta de hueso. Entre las estrategias de control respetuosas con el medio ambiente, cabe destacar el uso de agentes de biocontrol (ABCs). Esta tesis tiene como objetivo completar el desarrollo del ABC B. amyloliquefaciens CPA-8 para así obtener un producto eficaz que proporcione una estrategia comercialmente viable. Los resultado se clasifican en (i) caracterización de CPA-8 (diseño de marcadores moleculares, respuestas ecofisiológicas, sensibilidad a antibióticos, producción de enterotoxinas); (ii) desarrollo de productos basados en CPA-8 (mejora del medio de crecimiento, selección de la tecnología de formulación, optimización del secado por lecho fluido-atomización); y (iii) definición de la estrategia de control de CPA-8 (actividad en diferentes huéspedes, definición de umbrales técnicos y eficacia en ensayos de campo). La integración de los productos basados en CPA-8 en los sistemas de cultivo habituales contribuye en el manejo de las enfermedades de poscosecha en fruta de hueso en el marco de una agricultura sostenible y/o ecológica.
Brown rot caused by the fungus Monilinia spp. is responsible for substantial postharvest losses of stone fruit. Among the environment-friendly strategies of control, the application of biological control agents (BCAs), has been strongly considered. Therefore, this thesis aimed to complete de development of the BCA Bacillus amyloliquefaciens CPA-8 to obtain efficacious CPA-8-based products that provide a plausible commercial strategy. The main findings are classified into (i) CPA-8 characterisation (molecular markers design, ecophisiological responses, sensibility to antibiotics, production of enterotoxins); (ii) CPA-8-based products development (improvement of the growth medium, selection of the formulation approach, fluid-bed spray-drying optimisation); and (iii) definition of the CPA-8 control strategy (activity in a wide range of hosts, definition of some technical thresholds and field efficacy at harvest and postharvest time). The integration of the CPA-8-based products here developed into the usual cropping systems can contribute to the management of post-harvest diseases in stone fruit in the framework of a sustainable and/or organic agriculture.

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Budiharjo, Anto. "Plant-bacteria interactions." Doctoral thesis, Humboldt-Universität zu Berlin, Mathematisch-Naturwissenschaftliche Fakultät I, 2011. http://dx.doi.org/10.18452/16333.

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Bacillus amyloliquenaciense FZB42 ist ein bekanntes Pflanzenwachstum-stimulierendes Rhizobakterium. Es produziert neben einer Vielzahl an Sekundärmetaboliten mit antibakterieller und antifungaler Wirkung, auch das Pflanzenhormon IAA. Obwohl viele dieser Mechanismen diskutiert werden, ist wenig darüber bekannt, auf welche Weise die Bakterien das Pflanzenwachstum fördern. In dieser Arbeit wurde eine Transposonmutagenese mithilfe des ‘mariner-transposons’ durchgeführt, und so eine Transposonbibliothek erstellt. Diese wurde dann auf geeignete Phänotypen untersucht, um die Gene zu finden, welche bestimmte Phänotypen verursachen. So konnten drei Mutanten erzeugt werden, die auf Grund der gestörten Biofilmbildung und der Fähigkeit zu schwärmen die Pflanzenwurzeln nicht mehr kolonialisieren konnten. Eine solche degU-Mutante, welche in der Biofilmbildung und ‚Swarming’ defizitär war und zwei Mutanten (yusV und pabB), die eine Beeinträchtigung in der Biofilmbildung aufwiesen, konnten durch Komplementation und Retransformation bestätigt werden. Mithilfe des Lemna-Biosystems und anderer Analysen mit A. thaliana konnten drei Gene bei B. amyloliqufaciens FZB42 gefunden werden, die wichtig für die Förderung des Pflanzenwachstums sind. Koloniesierungsexperimente der Wurzeln von A. thaliana mit diesen Mutanten zeigten deutlich verändertes Wachstum, verglichen mit dem Wildtypstamm. Ein weiteres Ziel dieser Arbeit war es neue Antibiotika in Mutanten, die in ihren nicht-ribosomalen Synthesen blockiert sind, zu finden. So konnten durch die Untersuchungen der Transposonbibliothek der Mutanten zwei neue Antibiotika entdeckt werden. Genauere Analysen dieser Antibiotika bestätigten, dass es sich um ein neues Bacteriocin (Amylocyclicin A) und ein neues Thiazol/Oxazole-modifiziertes Microcin (Plantazolicin) handelt. Die abschließenden Arbeiten beschäftigten sich dann mit Untersuchungen von Genen, welche für die Produktion von Substanzen gegen Nematoden verantwortlich sind. Hierbei konnten vier Mutanten gefunden werden, die durch eine Transposoninsertion eine schlechtere.
Bacillus amyloliqufaciens FZB42 has been known as PGPR which has an impressive effect to improve plant growth. It produces not only vast array of secondary metabolites with antibacterial and antifungal activities, but also produces the plant hormone IAA. Although many mechanisms have been elucidated, our knowledge about basic molecular mechanisms responsible for its beneficial action is far from complete. In this study, transposon mutagenesis based on mariner tranposon was applied to generate tranposon library which then was screened to identify the genes involved in plant growth-promoting activity. Three mutants that were impaired in their ability to colonize plant surface due to defects in biofilm formation and swarming motility were found. One mutant (degU mutant) showed defect in biofilm formation and swarming motility, as well, two mutants (yusV mutant and pabB mutant) impaired in biofilm formation were confirmed by complementation and retransformation. Screening by the Lemna biosystem and further assays with A. thaliana revealed three genes responsible for reduction in plant growth promoting activity of B. amyloliqufaciens FZB42. Colonization studies of these mutants in A. thaliana roots revealed patterns different to the wild type. A further issue pursued in this study was to discover new antibiotics using a mutant which has been blocked in its nonribosomally pathway. Screening of tranposon librabries from this mutant led to the finding of two novel ribosomally synthesized antibiotics. Further characterization revealed that these new antibiotics belonged to a novel bacteriocin (Amylocyclicin A) and a novel thiazole/oxazole-modified microcin (Plantazolicin). Last work in this study was looking for genes responsible for nematocidal production. Four mutants which showed reduction in nematocidal activity due to transposon insertion were found.

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35

Budiharjo, Anto [Verfasser], Rainer [Akademischer Betreuer] Borriss, Thomas [Akademischer Betreuer] Börner, and Joachim [Akademischer Betreuer] Vater. "Plant-bacteria interactions : molecular mechanisms of phytostimulation by Bacillus amyloliquefaciens FZB42 / Anto Budiharjo. Gutachter: Rainer Borriss ; Thomas Börner ; Joachim Vater." Berlin : Humboldt Universität zu Berlin, Mathematisch-Naturwissenschaftliche Fakultät I, 2011. http://d-nb.info/1015081533/34.

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36

Hübert, Christine [Verfasser], and Wilhelm [Akademischer Betreuer] Jelkmann. "Einsatz von Erwinia tasmaniensis und Bacillus amyloliquefaciens als bakterielle Antagonisten zur biologischen Kontrolle des Feuerbrands / Christine Hübert ; Betreuer: Wilhelm Jelkmann." Heidelberg : Universitätsbibliothek Heidelberg, 2017. http://d-nb.info/1180740130/34.

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37

Scholz, Romy [Verfasser], Rainer [Akademischer Betreuer] Borriss, Joachim [Akademischer Betreuer] Vater, and Roderich [Akademischer Betreuer] SüßMuth. "Synthese der Bacteriocine Amylocyclicin A und Plantazolicin in Bacillus amyloliquefaciens FZB42 / Romy Scholz. Gutachter: Rainer Borriss ; Joachim Vater ; Roderich Süßmuth." Berlin : Humboldt Universität zu Berlin, Mathematisch-Naturwissenschaftliche Fakultät I, 2011. http://d-nb.info/1015016820/34.

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Koumoutsi, Alexandra [Verfasser], Joachim [Gutachter] Vater, and Rainer [Gutachter] Borriss. "Functional genome analysis of the plant-growth promoting bacterium Bacillus amyloliquefaciens strain FZB42 / Alexandra Koumoutsi ; Gutachter: Joachim Vater, Rainer Borriss." Berlin : Humboldt-Universität zu Berlin, 2007. http://d-nb.info/1208077279/34.

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39

Santos, Letícia Mileny. "Farelo de soja e farelo de milho como substratos na produção de amilase e protease por Bacillus amyloliquefaciens MO.04b." Universidade Estadual de Londrina. Centro de Ciências Exatas. Programa de Pós-Graduação em Biotecnologia, 2015. http://www.bibliotecadigital.uel.br/document/?code=vtls000203161.

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As enzimas microbianas amilase e protease são de grande importância biotecnológica e aplicáveis em diferentes setores industriais. Uma vez que resíduos gerados na agroindústria podem constituir problema ambiental, uma estratégia para seu aproveitamento é a fermentação em estado sólido (FES), processo potencial para produção de biomoléculas de interesse econômico, como enzimas. O objetivo do presente trabalho foi avaliar a produção de amilase e protease por Bacillus amyloliquefaciens MO.04b em farelo de soja (FS) e farelo de milho (FM). O inóculo de B. amyloliquefaciens foi desenvolvido em meio de sais mineriais (MSM) acrescido de glu 0,2 % por 24 h e 37 ˚C e seu volume foi avaliado entre 100, 200,300, 400 e 500 µL. As FES foram realizadas a partir de 1,5 g de FS e FM umedecidos com: água; MSM; MSM + glu 1 %; MSM + Extrato de levedura (YE) 1 % ou MSM + glu 1 %+ YE 1 %. A curva de crescimento em até 48 h foi desenvolvida tendo como resposta a atividade amilolítica e proteolitica, realizou-se ainda FES com misturas dos resíduos e a inserção do bagaço de cana-de-açúcar (BC) e a casca de arroz (CA) como agentes de estruturação. A atividade foi determinada pelas metodologias descritas por Miller (amilase) e Hartree (protease). A produção da enzima mais ativa foi otimizada através do delineamento de misturas simplex-centróide e parcialmente caracterizada nas condições ótimas. O melhor tempo para a obtenção do inóculo foi 18 h com 6,6 x 107 UFC/mL e a solução MSM + glu 1 %+ YE 1 % foi selecionada. As atividades nos extratos: FS 146,08 U/mL para protease e 75,63 U/mL para amilase e em FM 63,41 U/mL para protease e 0,752 U/mL para a amilase; ressaltando que essas atividades foram determinadas em extratos sem diálise (ESD). O melhor tempo de cultivo foi 36 h em ambos os substratos para a produção de amilase 1,36 U/mL em FS e 0,27 U/mL em FM e 9 h em FS para protease 11,7 U/mL, atividades essas determinadas em extratos dialisados (ED), e em FM não foi produzida protease. O agente de estruturação BC melhorou a atividade da amilase 3,62 U/mL e a CA melhorou a produção de protease 11,17 U/mL em FS. A condição ótima de produção de proteases, 20,32 U/mL, foi no substrato FS com casca de arroz como agente estruturante. A temperatura ótima 25 º C e o pH ótimo 3,0, além de estável por 90 min. a 70 º C mantendo 70 % de sua atividade original. Os resultados obtidos sugerem que esta protease apresenta propriedades interessantes para futuras aplicações biotecnológicas.
The microbial enzymes amylase and protease are of great biotechnological importance and applicable in different industrial sectors. Since waste generated in agribusiness may constitute environmental problem, a strategy for exploitation is the solid-state fermentation (SSF), potential process for the production of biomolecules of economic interest, such as enzymes. The objective of this study was evaluate the production of amylase and protease by B. amyloliquefaciens MO.04b in soybean meal and corn meal. The inoculum of B. amyloliquefaciens was developed in mineriais salts medium (MSM) plus 0.2% glu 37 ° C for 24 h and its volume was estimated at between 100, 200, 300, 400, and 500 µL. The SSF were made from 1.5 g of soybean meal and corn meal moistened with: water; MSM; MSM + 1% glu; MSM + yeast extract (YE) or 1% MSM + 1% glu + 1% YE. The growth curve within 48 h has been developed in response to amylolytic and proteolytic activity was also held-FES with mixtures of waste and the insertion of the cane bagasse (BC) and rice husk (CA) as bulking agents. The activity was determined by the methods described by Miller (amylase) and Hartree (protease). The production of the active enzyme was optimized through design of simplex-centroid mixtures and partially characterized in optimal conditions. The best time to obtain the 18 h inoculum was 6.6 x 107 CFU/mL and MSM + 1% glu + YE solution 1% was selected. The activity in the extracts soybean meal 146.08 U/mL for protease and 4.28 U/mL for amylase and corn meal 63.41 U/mL for protease and 0.752 U/mL for amylase; noting that these activities were determined in extracts without dialysis (ESD). The best culture time was 36 h in both substrates for the production of amylase 1.36 U/mL in soybean meal and 0.27 U/mL in corn meal for 9 h for protease in soybean meal to 11.7 U/mL, these activities determined in dialyzed extract (ED) and corn meal protease was not produced). The bulking agent BC improved amylase activity 3.62 U/mL, and the improved AC protease production 11.17 U/mL in soybean meal. The optimal condition for the production of proteases, 20.32 U/mL, was in soybean meal substrate with rice husk as bulking agent. The optimum temperature 25 ° C and the optimum pH 3.0, and stable for 90 min. to 70 ° C while maintaining 70% of its original activity. The results suggest that this protease has interesting properties for future biotechnological applications.

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Llario, Sempere Ferran. "IMPORTANCIA DE LOS MICROORGANISMOS EN LA PRODUCCIÓN DE LANGOSTINOS MARINOS EN SISTEMAS DE BIOFLÓCULOS Y SIN RENOVACIÓN DE AGUA." Doctoral thesis, Universitat Politècnica de València, 2018. http://hdl.handle.net/10251/111825.

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Los microorganismos son esenciales para poder llevar a cabo la producción intensiva de langostinos marinos, ya que son los encargados de mantener el agua con una calidad óptima para los animales. Además, los microorganismos permiten minimizar o eliminar las necesidades de realizar cambios de agua. Entre todos los microorganismos presentes en los sistemas intensivos, sin renovación de agua, las bacterias y las microalgas son los más abundantes y los que tienen un papel más importante en el sistema de producción. En esta tesis se profundiza en el conocimiento de la dinámica de las microalgas presentes en un sistema de preengorde y en un sistema de bioflóculos, para el cultivo intensivo de langostinos marinos, utilizando la metodología HPLC/CHEMTAX. Además, se testa si la bacteria Bacillus amyloliquefaciens puede tener un papel probiótico en los sistemas de bioflóculos y si es capaz de mejorar el proceso de maduración del sistema. Los resultados obtenidos han determinado que la metodología HPLC/CHEMTAX es una forma rápida y eficiente para analizar el fitoplancton y el perifiton en los sistemas intensivos de langostinos, siempre que predominen los procesos autotróficos. Esta técnica, ha permitido detectar por primera vez prasinofíceas y primnesiofíceas en el perifiton que crece sobre los tanques de policloruro de vinilo (PVC). También se ha observado la influencia del estado trófico del sistema y la exposición a la luz sobre la dinámica de las microalgas en los sistemas de bioflóculos. Por lo que respecta a la aplicación de Bacillus amyloliquefacien a los sistemas de bioflóculos, esta implica un refuerzo del sistema inmunológico de los langostinos, sumando sus efectos a los producidos por los bioflóculos. Estos beneficios, ocurren incluso al aplicar dosis del orden de 103 ufc/mL, siendo esta menor a la recomendada para otras bacterias probióticas. A pesar de que los resultados de otras investigaciones apuntaban que Bacillus amyloliquefaciens podría participar en la formación de bioflóculos, la mejora de la calidad del agua y el crecimiento de los langostinos, la aplicación de esporas sobre el agua no mostró efectos estadísticamente significativos sobre la dinámica del sistema, la calidad del agua o los parámetros zootécnicos.
Microorganisms are essential for the intensive production of marine shrimps, since they are responsible for maintaining the water with an optimum quality for them. Also, these microorganisms minimize or eliminate water renewal. Among all the microorganisms present in intensive culture systems, without water renewal, microalgae and bacteria play the most important role and are the most abundant ones. In this thesis we analyze the microalgae dynamics using HPLC/CHEMTAX methodology in two intensive culture systems of marine shrimp, a shrimp nursery and a biofloc system. We also test the role of the bacteria Bacillus amyloliquefaciens as a probiotic and its capacity to improve the maturation process of the biofloc system. The results showed that the HPLC/CHEMTAX methodology is a fast and efficient way to analyze phytoplankton and periphyton in intensive culture systems of shrimp, provided that autotrophic processes are predominant. This technique allowed to detect prasinophytes and prymnesiophytes for the first time in the periphyton that grows on the polyvinyl chloride (PVC) tanks. Also, it has allowed to study the influence of trophic state and light exposition on microalgae dynamic in the biofloc systems. The application of Bacillus amyloliquefaciens in biofloc systems strengthens the immune system of shrimp and adds its probiotic effects to that produced by bioflocs. These benefits happen even with doses of 103 cfu/mL, lower than recommended doses for other probiotic bacteria. Other researches indicated that Bacillus amyloliquefaciens might have other effects such as participating in the biofloc system formation, improving water quality and shrimp growth; however, the application of spores in the culture water did not show any statistically significant effect on the system dynamic, water quality or zootechnic parameters.
Els microorganismes són essencials per a poder dur a terme la producció intensiva de llagostins marins, ja que són els encarregats de mantenir l'aigua amb una qualitat òptima per als animals. A més a més, els microorganismes permeten minimitzar o eliminar les necessitats de realitzar canvis d'aigua. Entre tots els microorganismes presents en els sistemes intensius, sense renovació d'aigua, les bactèries i les microalgues són les més abundants i les que tenen un paper mes important en el sistema de producció. En aquesta tesis es profunditza en el coneixement de la dinàmica de les microalgues presents als sistemes de preengreixament i en un sistema de bioflòculs, per al cultiu intensiu de llagostins marins, utilitzant la metodologia HPLC/CHEMTAX. A més a més, es prova si la bactèria Bacillus amyloliquefaciens pot tenir un paper probiòtic en els sistemes de bioflòculs i si es capaç de millorar el procés de maduració del sistema. Els resultats obtinguts han determinat que la metodologia HPLC/CHEMTAX és una forma ràpida i eficient per a analitzar el fitoplàncton i el perifíton als sistemes intensius de llagostins, sempre que predominen els processos autotròfics. Aquesta tècnica, ha permès detectar per primera vegada prasinofícies i primnesiofícies al perifíton que creix sobre els tancs de policlorur de vinil (PVC). També s'ha observat l'influencia del estat tròfic del sistema i l'exposició a la llum sobre la dinàmica de les microalgues als sistemes de bioflòculs. Pel que respecta a l'aplicació de Bacillus amyloliquefaciens en els sistemes de bioflòculs, aquesta implica un reforç del sistema immunològic dels llagostins, sumant els seus efectes als produïts pels bioflòculs. Aquestos beneficis, permaneixer al aplicar dosis de l'ordre de 103 ufc/mL, sent aquesta menor a la recomanada per a altres bactèries probiòtiques. Encara que els resultats d'altres investigacions apunten que Bacillus amyloliquefaciens podria participar en la formació de bioflòculs, la millora de la qualitat de l'aigua i el creixement dels llagostins, l'aplicació d'espores a la columna d'aigua no va mostrar ningún efecte estadísticament significant sobre la dinàmica del sistema, la qualitat de l'aigua o els paràmetres zootècnics.
Llario Sempere, F. (2018). IMPORTANCIA DE LOS MICROORGANISMOS EN LA PRODUCCIÓN DE LANGOSTINOS MARINOS EN SISTEMAS DE BIOFLÓCULOS Y SIN RENOVACIÓN DE AGUA [Tesis doctoral no publicada]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/111825
TESIS

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Koumoutsi, Alexandra. "Functional genome analysis of the plant-growth promoting bacterium Bacillus amyloliquefaciens strain FZB42; characterizing its production and regulation of nonribosomal peptide synthetases." [S.l.] : [s.n.], 2006. http://deposit.ddb.de/cgi-bin/dokserv?idn=983473870.

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Mariappan, Aruljothi [Verfasser], Rainer [Akademischer Betreuer] Borriss, Joachim [Akademischer Betreuer] Vater, and Thomas [Akademischer Betreuer] Eitinger. "Molecular mechanisms controlling bacilysin biosynthesis in plant growth promoting rhizobacterium - Bacillus amyloliquefaciens FZB42 / Aruljothi Mariappan. Gutachter: Rainer Borriss ; Joachim Vater ; Thomas Eitinger." Berlin : Humboldt Universität zu Berlin, Mathematisch-Naturwissenschaftliche Fakultät I, 2012. http://d-nb.info/1025112407/34.

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Ziegler, Andreas [Verfasser], Ulf [Akademischer Betreuer] Stahl, and Matthias [Akademischer Betreuer] Rädle. "Bewegungsanalyse von Bacillus amyloliquefaciens durch Online-Mikroskopie mit automatisierter Wegverfolgung / Andreas Ziegler. Gutachter: Ulf Stahl ; Matthias Rädle. Betreuer: Ulf Stahl ; Matthias Rädle." Berlin : Technische Universität Berlin, 2015. http://d-nb.info/1071889885/34.

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Silva, Isadora Ferreira da. "Produ??o de amilase por Bacillus amyloliquefaciens utilizando torta de maca?ba (Acrocomia aculeata) e farinha de pupunha (Bactris gasipaes) como substratos." UFVJM, 2012. http://acervo.ufvjm.edu.br:8080/jspui/handle/1/501.

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Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico (CNPq)
Funda??o de Amparo ? Pesquisa do estado de Minas Gerais (FAPEMIG)
Amilases s?o enzimas que atuam no rompimento das liga??es glicos?dicas presentes nas cadeias de amilose e amilopectina. As amilases possuem aplica??es em diversas ?reas tais como: ind?stria de papel e celulose, ind?stria t?xtil, ind?stria de detergentes e produtos de limpeza, ind?stria qu?mica e farmac?utica, na produ??o de vitaminas e antibi?ticos. Embora as amilases sejam encontradas em plantas, animais e micro-organismos, as enzimas microbianas geralmente atendem de maneira satisfat?ria ? demanda industrial. As enzimas microbianas podem ser obtidas por meio de processos fermentativos, podendo ser obtidas tanto por cultivo superficial como por cultivos submersos. O presente trabalho teve por objetivo avaliar a produ??o de enzimas amilol?ticas por fermenta??o submersa utilizando Bacillus amyloliquefaciens ATCC 23350 e coprodutos da cadeia produtiva do biodiesel como substrato. A capacidade da produ??o das enzimas amilol?ticas foi testada utilizando como substratos diferentes coprodutos, sendo esses: torta de maca?ba, torta de dend?, farinha de pupunha, torta de mamona e torta de tremo?o. A torta de maca?ba e farinha de pupunha foram os substratos que demonstraram maior capacidade de produ??o das enzimas amilol?ticas, sendo os coprodutos selecionados para a etapa de otimiza??o da produ??o das enzimas. Diferentes condi??es de processo foram otimizadas com intuito de obter o m?ximo rendimento da produ??o de amilases, atrav?s do emprego de delineamentos experimentais aplicados ? Metodologia de Superf?cie de Resposta. Os processos fermentativos deste trabalho foram realizados ? temperatura de 37?C, 120 rpm de agita??o, utilizando solu??o de sais para suplementa??o do meio. As vari?veis pH, raz?o s?lido-l?quido (S/L) (% m/v) e concentra??o de NH4NO3 (gL-1) foram estudadas. As dosagens das atividades enzim?ticas foram realizadas em tempos de fermenta??o de 12, 24, 36, 48 e 72 horas. A m?xima produ??o das amilases foi alcan?ada com 24 horas de fermenta??o, S/L de 12% e pH 7,0, obtendo valores de 3.393 U.mL-1 com a farinha de pupunha e 196 U.mL-1 com a torta de maca?ba como substratos.
Disserta??o (Mestrado) ? Programa de P?s-Gradua??o em Qu?mica, Universidade Federal dos Vales do Jequitinhonha e Mucuri, 2012.
ABSTRACT Amylases are enzymes that act in the rupture of glycosidic bonds present in the chains of amylose and amylopectin. Amylases have applications in diverse areas such as pulp and paper industry, textiles, laundry detergents and cleaning products, chemical and pharmaceutical industry, production of vitamins and antibiotics. Although the amylases are found in plants, animals and micro-organisms, microbial enzymes generally meet satisfactorily the industrial demand. The microbial enzymes can be obtained by fermentative procedures, which may be obtained by cultivation both surface and submerged cultivation of micro-organisms. This study aimed to evaluate the production of amylolytic enzymes by submerged fermentation using Bacillus amyloliquefaciens ATCC 23350, and co-products from biodiesel production chain as substrates. The capacity of the production of amylolytic enzymes was tested using different substrates such as co-products, these being: macauba cake, palm kernel cake, peach palm flour, castor bean and lupine cake. The peach palm flour and macauba cake were the substrates which show greater production capacity of amylolytic enzymes, and co-products selected for the optimization step of enzyme production. Different process conditions were optimized with the aim of obtaining the maximum yield of amylases, through the use of experimental designs applied to the Response Surface Methodology. The fermentation process of this work was performed at 37?C, 120 rpm agitation, using salt solution for supplementation of the medium. The pH, liquid-solid ratio (S/L) (% w/v) and concentration of NH4NO3 (gl-1) were studied. The measurements of enzyme activities were carried out in fermentation times of 12, 24, 36, 48 and 72 hours. Maximum production of amylase was achieved with 24 hours of fermentation, S/L of 12% and pH 7.0 to obtain values of 3.393 U.mL-1 with peach palm flour and 196 U.mL-1 with maca?ba cake as substrate.

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Thammakulkrajang, Rarinthorn. "Pressure-Assisted Thermal Processing of Bacterial Spores: Influence of Selected Product and Packaging Parameters." The Ohio State University, 2018. http://rave.ohiolink.edu/etdc/view?acc_num=osu1532014631156644.

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Tian, Feng. "Enzymatic synthesis of fructooligosaccharides and levan through transfructosylation reaction catalyzed by levansucrase from Bacillus Amyloliquefaciens and by its combined use with endo-inulinase." Thesis, McGill University, 2013. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=119466.

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Both intracellular and extracellular levansucrase (LS) were recovered from Bacillus amyloliquefaciens biomass. Sucrose was identified as an important inducer for the expression of LSs. Polyethylene glycol (PEG 200) selectively fractionated intracellular (vs. extracellular) LS activity. Partially purified intra- and extracellular LSs were studied in terms of their optimum reaction temperature, pH, kinetics as well as their catalytic properties. B. amyloliquefaciens LSs exhibited high levan-forming activity over a wide range of sucrose concentrations. The optimum temperatures for the intra- (25-30 °C) and extracellular (40 °C) LS transfructosylation activities were lower than those for their hydrolytic activities (45-50°C). Intracellular LS's catalytic efficiency in transfructosylation exceeded that of extracellular LS. Intracellular LS was purified to homogeneity by size-exclusion chromatography and its catalytic properties was determined. The purified LS's transfructosylic activity (kcatapp of 1136.5 s−1) exceeded its hydrolytic activity. The kinetics of LS's transfructosylation activity were best fitted to the Hill model. In LS donor specificity studies, a shift of the transfructosylation reaction further towards the polymerization was observed when raffinose was used as fructosyl donor as compared to sucrose. In fructooligosaccharide (FOS) synthesis, disaccharides were more favourable fructosyl acceptors than monosaccharides. However, a lower accumulation of levan was detected in the maltose/sucrose reaction, and a greater amount of erlose was the dominant transfructosylation product in the reaction. This study is the first to highlight the catalytic efficiency of B amyloliquefaciens LS to synthesize a variety of hetero-FOSs from various saccharides, with lactose and maltose being the best fructosyl acceptors. Purified intracellular LS was subsequently combined with fructanases from Aspergillus niger to achieve the one-step synthesis of controlled-size novel FOSs and oligolevans from sucrose. Endo-inulinase from A. niger was able to hydrolyze both low and high MW levan and thus gave rise to different MW FOSs, whilst fructanase® (endo- and exo-inulinases) hydrolyzed levans almost completely to fructose. Therefore, endo-inulinase was chosen for use in the combined bi-enzymatic reactions. In this novel biocatalytic system, LS mainly catalyzed the synthesis of oligolevan and levan, whilst endo-inulinase regulated the product molecular size and acceptor availability. The novel bi-enzymatic system showed higher yield and productivity (67% w/w; 96 g/L/h) of transfructosylation products, FOSs and oligolevans, than the LS enzymatic system (3.0% w/w; 0.8 g/L.h) alone. The contribution of endo-inulinase's hydrolytic activity to the formation of short chain FOS (scFOS) and oligolevans was greater than that of LS through its acceptor reaction; however, the production of intermediate levans with appropriate MW by LS was the prerequisite. The effects of sucrose concentration, reaction time, and enzymatic ratio on FOSs, oligolevans, and total transfructosylation products were assessed through a response surface methodology (RSM). Additional experimental runs using the novel bi-enzymatic system confirmed the RSM-predicted maximal FOS and oligolevan levels. RSM analysis showed sucrose level to be the most influential of the design variables tested, whilst the LS to endo-inulinase ratio had significant effects on levanohexaose and oligolevans formation. Given the biocatalytic system's low reaction temperatures (35 °C) and substrate concentrations (0.6 M), the novel biocatalytic system showed great potential for industrial applications, where huge cost effectiveness benefits would arise from commercial scale-up production. In addition, the short reaction time as well as high FOSs yield of the novel biocatalytic system are also superior to all current documented FOSs biocatalytic systems.
L'enzyme levansucrase intracellulaire et celle extracellulaire ont été isolées de la biomasse du Bacillus amyloliquefaciens. Le sucrose a été identifié comme un inducteur important pour l'expression de la levansucrase. Les levansucrases intra- et extracellulaire, purifiées partiellement, ont été caractérisées en termes de température optimale, pH optimal, paramètres cinétiques, et propriétés catalytiques. Les levansucrases de B. amyloliquefaciens ont démontré une activité catalytique élevée pour la formation de levan à travers une large gamme de concentrations du sucrose. Les températures optimales des activités de transfructosylation des levansucrases intracellulaire (25-30°C) et extracellulaire (40ºC) sont plus faibles que celles de leur activités hydrolytiques (45-50°C). L'efficacité catalytique de la levansucrase intracellulaire en transfructosylation a dépassé celle de la levansucrase extracellulaire. La levansucrase intracellulaire a été purifiée jusqu'à l'homogénéité par chromatographie d'exclusion stérique et ses propriétés catalytiques ont été déterminées. L'activité de transfructosylation de la levansucrase purifiée (kcatapp de 1136.5 s−1) a été plus élevée que son activité hydrolytique. Les cinétiques catalytiques de l'activité de transfructosylation ont été mieux décrites par le modèle de Hill. L'étude de la spécificité de la levansucrase a démontré une orientation de la réaction de transfructosylation vers la polymérisation quand le raffinose a été utilisé comme donneur de fructose. Durant la synthèse du fructooligosaccharides, les disaccharides ont été favorisés comme accepteurs, par rapport aux monosaccharides. Cependant, une accumulation moins importante du levan a été obtenue dans la réaction de transfructosylation utilisant maltose et sucrose comme substrats. Cette étude est la première à démontrer l'efficacité catalytique de la levansucrase de B. amyloliquefaciens pour la synthèse de divers FOS à partir de saccharides variés, dont le lactose et le maltose ont été les meilleurs substrats accepteurs de fructose. La levansucrase intracellulaire purifiée a été combinée avec l'endo-inulinase de Aspergillus niger dans une seule étape réactionnelle pour synthétiser de nouveaux FOSs et des oligolevans de tailles contrôlées à partir du sucrose. Dans ce nouveau système bi-enzymatique, la levansucrase a surtout catalysé la formation d'oligolevan et de levan, tandis que l'endo-inulinase a régulé la taille moléculaire des produits et la disponibilité des accepteurs. Le nouveau système bi-enzymatique a démontré un rendement et une productivité (67% m/m; 96 g/L/h) plus élevés des produits de transfructosylation, des FOS et des oligolevans, que le système mono-enzymatique (3.0% m/m; 0.8 g/L.h). La contribution de l'activité hydrolytique de l'endo-inulinase dans la formation des courtes chaînes de FOS et des oligolevans a été plus importante que celle de la levansucrase. Les effets de la concentration du sucrose, du temps de réaction, et du ratio enzymatique sur les rendements des FOSs, des oligolevans, et des produits de transfructosylation en général ont été étudiés à travers la méthode des surfaces de réponses. Les résultats expérimentaux ont confirmé et validé les prédictions des modèles développés. La même analyse a démontré que le niveau du sucrose est la variable la plus influante parmi les variables évaluées, tandis que le ratio de la levansucrase et de l'endo-inulinase a eu un impact important sur la synthèse de levanohexaose et des oligolevans. Étant donné que la température de la réaction biocatalytique et les concentrations des substrats sont faibles, ce nouveau système bi-enzymatique démontre un grand potentiel pour les applications au niveau industriel. Par ailleurs, le temps de la réaction plutôt bref et les rendements élevés des FOSs démontrent la supériorité du système bi-enzymatique développé en comparaison avec les systèmes biocatalytiques actuels qui ont été documentés.

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Deravel, Jovana. "Caractérisation des lipopeptides d’origine non ribosomique comme biopesticides." Thesis, Lille 1, 2011. http://www.theses.fr/2011LIL10126.

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Le premier objectif de ce travail a été d’étudier l’implication des lipopeptides de Bacillus spp. dans la colonisation de la rhizosphère de tomates. Alors que seules les souches produisant de la surfactine sont capables de coloniser un milieu synthétique, toutes les souches testées, colonisent la rhizosphère de tomates avec une plus ou moins bonne efficacité quelque soit leur propriété de production de lipopeptide(s). L’efficacité de la colonisation des racines de tomates est principalement espèce-dépendante. Ce n’est que quand une souche est déjà une bonne colonisatrice que la surfactine semble améliorer cette propriété. Le deuxième objectif a été de tester l’effet des lipopeptides surfactine et mycosubtiline contre le phytopathogène obligatoire de la laitue : Bremia lactucae. À l’échelle du laboratoire, la mycosubtiline à 100 mg/L réduit le pourcentage de plantes infestées de 70 %. La surfactine ne montre aucun effet contre le champignon. Un mélange de mycosubtiline et de surfactine à 50 mg/L chacun diminue le pourcentage de plantes infestées de 65 %. Il semble diminuer le nombre de spores par plante infestée alors que cette propriété n’est pas remarquée avec les autres traitements. L’utilisation de la mycosubtiline dans une serre de culture limite la maladie aux symptômes les moins sévères et protège les plantes saines d’une contamination croisée. L’action des lipopeptides dans la colonisation des racines par Bacillus spp. n’avait jamais été validée in situ. De même, c’est la première fois que l’activité des lipopeptides est testée contre un phytopathogène obligatoire
The first aim of this work was to study the role played by the lipopeptides of Bacillus spp. in the colonization of the tomato rhizosphere. While only the strains producing surfactin are able to colonize a synthetic agar medium, all the strains are able to colonize the rhizosphere of tomatoes with a more or less good efficiency, whatever the lipopeptide(s) they have the capability to produce. The efficiency of the colonization of the tomato rhizosphere is species-dependant. However, surfactine seems to improve the efficiency of only the good colonizing strains. The second aim of this thesis was to test the effect of surfactin and mycosubtilin against a biotrophic parasite of lettuce: Bremia lactucae. Used at 100 mg/L, mycosubtilin reduces the percentage of infested plants of 70 %. Surfactin does not have effect against the fungy. A mixture of mycosubtilin and surfactin at both 50 mg/L decreases the percentage of infested plants of 65 %. This mixture seems to reduce the number of spore per infested plant while this property was not found for the other treatments. The use of mycosubtilin in a greenhouse confines the disease to the lowest classes of severity and protects the healthy plants from a cross contamination.The efficiency of lipopeptides of Bacillus spp. in root colonization by these bacteria was never tested in situ before. Furthermore, this is the first time that the activity of lipopeptides is validated against an obligate phytopathogen

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Huang, Hsiang-yan, and 黃湘燕. "The study of keratinase from Bacillus amyloliquefaciens." Thesis, 2016. http://ndltd.ncl.edu.tw/handle/24079766033851852039.

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碩士
國立臺灣大學
生化科技學系
104
Feather is composed of keratin, with more than 80% amino acid. According to research, keratin is difficult to degrade. Strong acid and basic chemical solvent is used on industry, which will be easily degrade feather. Some factory boil and grind feather into powder, and use it as additive fodder. But there should be more usage by feather. Since 1950, more research study about degrading feather by biodegradation. It is not only a better way to degrade feather but also maintains ingredient and function of feather. And some research also found that feather-degrading enzyme with other function. So we isolated bacterial strain from environment and have cloned a functional gene of keratinase. Then we purified the enzyme to study feature of the enzyme. In our study, after 16s rRNA sequencing, isolated strains were confirmed as Bacillus cereus and Bacillus amyloliquefaciens, and identified as Bacillus cereus Ker103 and Bacillus amyloliquefaciens Ker103. We produced enzyme by directly culture and Escherichia coli BL21 expression system, then analyzed enzyme activity and stability. Between these two methods, purity, productivity and activity was different. Enzymes produced from these two strains are stable and tolerated in different environment. Under pH 3.0-10.0 and 20-70℃, the two enzymes keep their activity and stability. Crude enzyme produced from Bacillus cereus Ker103 indicated the best activities are under incubation condition pH 8.0 and 50℃. The crude enzyme produced from Bacillus amyloliquefaciens Ker103 show the best activity under pH 7.0 and 40℃. We use PCR to detect and clone the enzyme gene. Bacillus cereus Ker103 includes enzyme gene, vpr, with predicted molecular weight 98.9 kDa and Bacillus amyloliquefaciens Ker103 includes enzyme gene, kerk, with predicted molecular weight 28.8 kDa. From the result of SDS-PAGE of these two cultural supernatant, indicated that these two protein are extracellular enzymes. Among these two enzymes, Vpr formed inclusion body, which made it difficult to produce and purify. So we especially try to purify KerK protein by Escherichia coli BL21 expression system to improve its purity. Then we detected enzyme activity, we found that purified enzyme show the best activity under pH 9.0, easily get lost activity under high salt concentration condition. The addition of surfactant Tween 20 and PEG-3350 improve the enzyme activity. We concluded that KerK showed its potential on large scale production and application.

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49

Huang, Yaohua. "Non-thermal Plasma Inactivation of Bacillus Amyloliquefaciens spores." 2011. http://trace.tennessee.edu/utk_gradthes/980.

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Bacterial spores have remarkable resistance to a variety of harsh conditions, causing spoilage in food industry and becoming the primary bacterial agent in biowarfare and bioterrorism. In this study, inactivation mechanisms of Bacillus amyloliquefaciens (BA) spores by non-thermal plasma (NTP) were investigated by using Fourier-transform infrared spectroscopy (FTIR) as a major tool to exam spores after NTP treatment. Chemometric techniques, such as multivariate classification models based on soft independent modeling of Class Analogy (SIMCA) and Principal Component Analysis (PCA), were employed to identify functional group changes in FTIR spectra. The IR absorbance bands correlated to dipicolinic acid (DPA) decreased after NTP treatment indicating that DPA released and then reacted with reactive species generated by NTP and it was confirmed by nuclear magnetic resonance (NMR). Also IR absorbance bands corresponding to protein structure changed. FTIR combined with UV-Vis spectroscopy was used to monitor spore germination. Large amount of DPA released in a short time when spores germinated at 50°C, showing that DPA released in response to heating. NTP treated spores could germinate with little DPA release due to sub-lethal effects induced by plasma. Also an empirical model based on Weibull distribution was established to describe the spore germination process showing that NTP treated spores exhibited abnormal germination pattern. Inactivation mechanisms of NTP with air as feed gas was compared with high-pressure, wet heat, chemical treatment using chlorine dioxide (CD) and NTP with argon as feed gas. The results showed that few chemical changes in spores after autoclave and high pressure treatments, though protein structure changed. CD and NTP with air as feed gas inactivated spores by oxidation. DPA released after NTP with argon as feed gas treatment and it is possible that UV and charged particles accounts for the inactivation. This study provides in depth insight into the inactivation mechanism of NTP and information for optimizing NTP process.

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50

Shih, Po-Shu, and 施博書. "Purification and characterization of LipA from Bacillus amyloliquefaciens." Thesis, 2018. http://ndltd.ncl.edu.tw/handle/58ww9c.

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碩士
國立臺灣大學
農業化學研究所
106
Microbial lipases are mainly applied in food industry, detergents and the other chemical industry. Because the isolation and purification of lipases from microorganisms is easier than those of animals and plants, the mass production and study of lipases from microorganisms are really important. Lipases of Bacillus spp. are heat and alkaline tolerant, so the lipase activity under heat and alkaline conditions can remain stable. Besides, lipases from the Bacillus spp. belong to those of the smallest molecular weight, and this make it a benefit for application in biotechnology. However, the original bacterial strain usually can not produce large amount of lipases, so it is better to select a transformant carrying a lipase gene in a plasmid which can produce large amount of lipases with special and specific characteristics for effective application in industry. Although many lipase genes form Bacillus spp. have been sequenced, studies on lipases of Bacillus amyloliquefaciens are still not comprehensive. A strain of Bacillus amyloliquefaciens was isolated from the environment and its lipase gene was selected, and cloned into pET21a, for expressing in E.coli by an induction with IPTG. Through the purification of the lipase protein (LipA) by the nickle column, a protein whose molecular weight is about 25kDa was obtained. The optimal temperature and pH value for LipA lipase activity are 35℃ and pH-10, respectively. Also, the LipA lipase is stable under 35℃ and pH value 9-10. The LipA lipase prefers p-nitrophenyl laurate as its substrate. The buffer containing a final concentration of 10 mM Zn2+, Fe2+, Cu2+, Ca2+, SDS or Triton-X-100 would strongly suppress the enzymatic activity. 10% DMSO and isopropanol would slightly enhance the enzyme activity, but 25% of most organic solvent could inhibit LipA activity. The results of the thin-layer-chromatography (TLC) showed that the longer the incubation time, the more degradation product of olive oil, including fatty acid and diglyceride. The degradation rate of olive oil after 96 hrs by B. amyloliquefaciens, the transformant expressing and LipA lipase was 38.8%, 45.3% and 53.5%, respectively, whereas for soybean oil, it was about 35.5%, 43% and 62%, respectively. Taken together, the above results revealed that LipA lipase has the potential to be applied in biodiesel production, recycled oil treatment and oil modification.

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