Добірка наукової літератури з теми "Extracellular hydrolytic activities"

Biotransformation via solid state fermentation (SSF) mediated by microorganisms is a promising approach to produce useful products from agricultural biomass. Lactic acid bacteria (LAB) that are commonly found in fermented foods have been shown to exhibit extracellular proteolytic, β-glucosidase, β-mannosidase, and β-mannanase activities. Therefore, extracellular proteolytic, cellulolytic, and hemicellulolytic enzyme activities of seven Lactobacillus plantarum strains (a prominent species of LAB) isolated from Malaysian foods were compared in this study. The biotransformation of palm kernel cake (PKC) biomass mediated by selected L. plantarum strains was subsequently conducted. The results obtained in this study exhibited the studied L. plantarum strains produced versatile multi extracellular hydrolytic enzyme activities that were active from acidic to alkaline pH conditions. The highest total score of extracellular hydrolytic enzyme activities were recorded by L. plantarum RI11, L. plantarum RG11, and L. plantarum RG14. Therefore, they were selected for the subsequent biotransformation of PKC biomass via SSF. The hydrolytic enzyme activities of treated PKC extract were compared for each sampling interval. The scanning electron microscopy analyses revealed the formation of extracellular matrices around L. plantarum strains attached to the surface of PKC biomass during SSF, inferring that the investigated L. plantarum strains have the capability to grow on PKC biomass and perform synergistic secretions of various extracellular proteolytic, cellulolytic, and hemicellulolytic enzymes that were essential for the effective biodegradation of PKC. The substantial growth of selected L. plamtraum strains on PKC during SSF revealed the promising application of selected L. plantarum strains as a biotransformation agent for cellulosic biomass.

Plasma membrane vesicles were purified from wheat roots by discontinuous sucrose gradient centrifugation and two-phase partitioning methods. The influence of extracellular-side Ca2+ on the activity of the plasma membrane H+ -ATPase from wheat roots was studied. The results showed that the ATP hydrolytic activities of the plasma membrane H+ -ATPase were inhibited by the cytoplasmic-side Ca2+. Within 0~200 µmol/L the ATPase activity decreased gradually with the increase in Ca2+ concentration; the ATPase activity was inhibited by 40% when Ca2+ concentration was 1000 µmol/L. However, the ATP hydrolytic activities were recovered by the presence of extracellular-side Ca2+. Results showed that the ATPase activities were increased with the increase in extracellular-side Ca2+; when the extracellular-side Ca2+ was 1000 µmol/L, the ATPase activity was recovered by 87.5%. Further studies found that the extracellular-side Ca2+ increased the DPH polarisation and decreased the MC540 fluorescence intensity, showing that membrane fluidity was decreased and membrane stacking was increased by the external Ca2+. The above results suggested that the plasma membrane H+ -ATPase could be regulated by the extracellular side Ca2+ through affecting the plasma membrane physical states.

Abstract. Marine snow aggregates are heavily colonized by heterotrophic microorganisms that express high levels of hydrolytic activities, making aggregates hotspots for carbon remineralization in the ocean. To assess how aggregate formation influences the ability of seawater microbial communities to access organic carbon, we compared hydrolysis rates of six polysaccharides in coastal seawater after aggregates had been formed (via incubation on a roller table) with hydrolysis rates in seawater from the same site that had not incubated on a roller table (referred to as whole seawater). Hydrolysis rates in the aggregates themselves were up to three orders of magnitude higher on a volume basis than in whole seawater. The enhancement of enzyme activity in aggregates relative to whole seawater differed by substrate, suggesting that the enhancement was under cellular control, rather than due to factors such as lysis or grazing. A comparison of hydrolysis rates in whole seawater with those in aggregate-free seawater, i.e. the fraction of water from the roller bottles that did not contain aggregates, demonstrated a nuanced microbial response to aggregate formation. Activities of laminarinase and xylanase enzymes in aggregate-free seawater were higher than in whole seawater, while activities of chondroitin, fucoidan, and arabinogalactan hydrolyzing enzymes were lower than in whole seawater. These data suggest that aggregate formation enhanced production of laminarinase and xylanase enzymes, and the enhancement also affected the surrounding seawater. Decreased activities of chondroitin, fucoidan, and arabinoglactan-hydrolyzing enzymes in aggregate-free seawater relative to whole seawater are likely due to shifts in enzyme production by the aggregate-associated community, coupled with the effects of enzyme degradation. Enhanced activities of laminarin- and xylan-hydrolyzing enzymes in aggregate-free seawater were due at least in part to cell-free enzymes. Measurements of enzyme lifetime using commercial enzymes suggest that hydrolytic lifetimes of cell-free enzymes may be sufficiently long to affect carbon remineralization in areas far from their site of production. Aggregate formation may be an important mechanism shaping the spectrum of enzymes active in the ocean, stimulating production of cell-free enzymes and leading to spatial and temporal decoupling of enzyme activity from the microorganisms that produced them.

Abstract. Marine snow aggregates are heavily colonized by heterotrophic microorganisms that express high levels of hydrolytic activities, making aggregates hotspots for carbon remineralization in the ocean. To assess how aggregate formation influences the ability of seawater microbial communities to access organic carbon, we compared hydrolysis rates of six polysaccharides in coastal seawater after aggregates had been formed (via incubation on a roller table) with hydrolysis rates in seawater from the same site that had not incubated on a roller table (referred to as whole seawater). Hydrolysis rates in the aggregates themselves were up to three orders of magnitude higher on a volume basis than in whole seawater. The enhancement of enzyme activity in aggregates relative to whole seawater differed by substrate, suggesting that the enhancement was under cellular control, rather than due to factors such as lysis or grazing. A comparison of hydrolysis rates in whole seawater with those in aggregate-free seawater, i.e. the fraction of water from the roller bottles that did not contain aggregates, demonstrated a nuanced microbial response to aggregate formation. Activities of laminarinase and xylanase enzymes in aggregate-free seawater were higher than in whole seawater, while activities of chondroitin, fucoidan, and arabinogalactan hydrolyzing enzymes were lower than in whole seawater. These data suggest that aggregate formation enhanced production of laminarinase and xylanase enzymes, and the enhancement also affected the surrounding seawater. Decreased activities of chondroitin, fucoidan, and arabinoglactan-hydrolyzing enzymes in aggregate-free seawaters relative to whole seawater are likely due to shifts in enzyme production by the aggregate-associated community, coupled with the effects of enzyme degradation. Enhanced activities of laminarin- and xylan-hydrolyzing enzymes in aggregate-free seawater were due at least in part to cell-free enzymes. Measurements of enzyme lifetime using commercial enzymes suggest that hydrolytic cell-free enzymes may be active over timescales of days to weeks. Considering water residence times of up to 10 days in the investigation area (Apalachicola Bay), enzymes released from aggregates may be active over timescales long enough to affect carbon cycling in the Bay as well as in the adjacent Gulf of Mexico. Aggregate formation may thus be an important mechanism shaping the spectrum of enzymes active in the ocean, stimulating production of cell-free enzymes and leading to spatial and temporal decoupling of enzyme activity from the microorganisms that produced them.

AbstractCold−adapted marine bacteria producing extracellular hydrolytic enzymes are important for their industrial application and play a key role in degradation of particulate organic matter in their natural environment. In this work, members of a previously−obtained protease−producing bacterial collection isolated from different marine sources from Potter Cove (King George Island, South Shetlands) were taxonomically identified and screened for their ability to produce other economically relevant enzymes. Eighty−eight proteolytic bacterial isolates were grouped into 25 phylotypes based on their Amplified Ribosomal DNA Restriction Analysis profiles. The sequencing of the 16S rRNA genes from representative isolates of the phylotypes showed that the predominant culturable protease−producing bacteria belonged to the class Gammaproteobacteria and were affiliated to the genera Pseudomonas, Shewanella, Colwellia, and Pseudoalteromonas, the latter being the predominant group (64% of isolates). In addition, members of the classes Actinobacteria, Bacilli and Flavobacteria were found. Among the 88 isolates screened we detected producers of amylases (21), pectinases (67), cellulases (53), CM−cellulases (68), xylanases (55) and agarases (57). More than 85% of the isolates showed at least one of the extracellular enzymatic activities tested, with some of them producing up to six extracellular enzymes. Our results confirmed that using selective conditions to isolate producers of one extracellular enzyme activity increases the probability of recovering bacteria that will also produce additional extracellular enzymes. This finding establishes a starting point for future programs oriented to the prospecting for biomolecules in Antarctica.

Actinomycetes has large habitats and can be isolated from terrestrial soil, rhizospheres of plant roots, and marine sediments. Actinomycetes produce several bioactive secondary metabolites with antibacterial, antifungal, and antiviral properties. In this study, some Actinomycetes strains were isolated from the rhizosphere zone of four different plant species: rosemary, acacia, strawberry, and olive. The antagonistic activity of all isolates was screened in vitro against Escherichia coli and Bacillus megaterium. Isolates with the strongest bioactivity potential were selected and molecularly identified as Streptomyces sp., Streptomyces atratus, and Arthrobacter humicola. The growth-promoting activity of the selected Actinomycetes isolates was in vivo evaluated on tomato plants and for disease control against Sclerotinia sclerotiorum. The results demonstrated that all bacterized plants with the studied Actinomycetes isolates were able to promote the tomato seedlings’ growth, showing high values of ecophysiological parameters. In particular, the bacterized seedlings with Streptomyces sp. and A. humicola showed low disease incidence of S. sclerotiorum infection (0.3% and 0.2%, respectively), whereas those bacterized with S. atratus showed a moderate disease incidence (7.6%) compared with the positive control (36.8%). In addition, the ability of the studied Actinomycetes to produce extracellular hydrolytic enzymes was verified. The results showed that A. humicola was able to produce chitinase, glucanase, and protease, whereas Streptomyces sp. and S. atratus produced amylase and pectinase at high and moderate levels, respectively. This study highlights the value of the studied isolates in providing bioactive metabolites and extracellular hydrolytic enzymes, indicating their potential application as fungal-biocontrol agents.

Les bactéries hétérotrophes jouent un rôle primordial dans le fonctionnement des écosystèmes marins. Cette thèse s’est intéressée aux facteurs influençant la composition des communautés bactériennes estuariennes et marines ainsi que leurs fonctions dans la dégradation de la matière organique. Pour cela, une étude a été réalisée au sein de l’estuaire de l’Aulne et des eaux côtières adjacentes de la rade de Brest. Des expériences en laboratoire ont permis d’appréhender le rôle spécifique des communications bactériennes basées sur le quorum sensing (QS). Les résultats obtenus ont montré que l’estuaire de l’Aulne était un véritable bioréacteur naturel. Il abrite des communautés bactériennes dont la composition et le niveau d’activité enzymatique suivent de fortes variations spatiotemporelles, liées aux conditions physicochimiques mais également à des facteurs biotiques comme leur degré de concurrence.Ils ont également démontré la forte implication du QS dans la régulation de la synthèse d’enzymes hydrolytiques au sein de souches isolées et de communautés bactériennes naturelles. Ce mécanisme de régulation peut aussi impacter la composition des communautés bactériennes en régulant les processus liés à la colonisation et aux interactions microbiennes, qu’elles soient trophiques ou non. Dans son ensemble, ce travail de thèse met en avant l’importance de considérer les facteurs biotiques dans l’étude des communautés bactériennes naturelles. Il souligne aussi la nécessité de mieux caractériser les interactions microbiennes, de leur mise en place d’un point de vue moléculaire à leurs impacts biogéochimiques globaux au sein des écosystèmes marins
Heterotrophic bacterial communities play a crucial role in marine ecosystems. This thesis addressed the factors influencing marine and estuarine bacterial community composition and function in organic matter degradation. To this end, we performed both in situ monitoring of the Aulne estuary and the adjacent coastal waters of the Bay of Brest, as well as laboratory experiments investigating the specific role of quorum sensing (QS), a bacterial communication system. Our results showed that the Aulne estuary acted as a natural bioreactor, fostering bacterial communities whose composition and intense hydrolytic enzymatic levels exhibited sharp spatiotemporal variations. These patterns were linked to variations in physicochemical conditions within the estuary, but also in biotic factors such as community composition or co-occurrence level. In addition, our results demonstrated that QS was greatly involved in the regulation of hydrolytic enzymes synthesis among isolated bacterial strains and natural bacterial communities. This mechanism also impacted bacterial community composition, likely by regulating processes involved in colonization or bacterial interactions. Altogether, this thesis highlighted the importance of considering biotic factors in the study of estuarine and marine bacterial communities. It also revealed the need to better characterize bacterial interactions, from their molecular implementation to their global biogeochemical impacts in marine ecosystems