Relevant bibliographies by topics / Enzymes Separation

Academic literature on the topic 'Enzymes Separation'

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Published: 10 December 2022
Last updated: 29 January 2023

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Journal articles on the topic "Enzymes Separation"

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Lemmer, Balázs, Szabolcs Kertész, Gábor Keszthelyi-Szabó, Kerime Özel, and Cecilia Hodúr. "Sonicated membrane separation." Progress in Agricultural Engineering Sciences 14, s1 (July 2018): 89–99. http://dx.doi.org/10.1556/446.14.2018.s1.9.

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Membrane separation processes are currently proven technologies in many areas. The main limitation of these processes is the accumulation of matter at the membrane surface which leads to two phenomena: concentration polarization and membrane fouling. According to the publications of numerous authors permeate flux could be increased by sonication. Our work focuses on separation of real broth by sonicated ultrafiltration. The broth was originated from hydrolysis of grounded corn-cob by xylanase enzyme. The filtration was carried out in a laboratory batch stirred cell with a sonication rod sonicator. In our work the effect of the stirring, the intensity of sonication and the membrane-transducer distance was studied on the efficiency of the ultrafiltration and on the quality of separated enzymes. Results reveal that xylanase enzyme can be effectively separated from real fermentation broth by ultrafiltration and enzymes keep their activity after the process. Enzyme activity tests show that low energy sonication is not harmful to the enzyme.
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Dey, Krishna Kanti, Sambeeta Das, Matthew F. Poyton, Samudra Sengupta, Peter J. Butler, Paul S. Cremer, and Ayusman Sen. "Chemotactic Separation of Enzymes." ACS Nano 8, no. 12 (October 2014): 11941–49. http://dx.doi.org/10.1021/nn504418u.

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Prouteau, Manoël, and Robbie Loewith. "Regulation of Cellular Metabolism through Phase Separation of Enzymes." Biomolecules 8, no. 4 (December 3, 2018): 160. http://dx.doi.org/10.3390/biom8040160.

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Metabolism is the sum of the life-giving chemical processes that occur within a cell. Proper regulation of these processes is essential for all organisms to thrive and prosper. When external factors are too extreme, or if internal regulation is corrupted through genetic or epigenetic changes, metabolic homeostasis is no longer achievable and diseases such as metabolic syndrome or cancer, aging, and, ultimately, death ensue. Metabolic reactions are catalyzed by proteins, and the in vitro kinetic properties of these enzymes have been studied by biochemists for many decades. These efforts led to the appreciation that enzyme activities can be acutely regulated and that this regulation is critical to metabolic homeostasis. Regulation can be mediated through allosteric interactions with metabolites themselves or via post-translational modifications triggered by intracellular signal transduction pathways. More recently, enzyme regulation has attracted the attention of cell biologists who noticed that change in growth conditions often triggers the condensation of diffusely localized enzymes into one or more discrete foci, easily visible by light microscopy. This reorganization from a soluble to a condensed state is best described as a phase separation. As summarized in this review, stimulus-induced phase separation has now been observed for dozens of enzymes suggesting that this could represent a widespread mode of activity regulation, rather than, or in addition to, a storage form of temporarily superfluous enzymes. Building on our recent structure determination of TOROIDs (TORc1 Organized in Inhibited Domain), the condensate formed by the protein kinase Target Of Rapamycin Complex 1 (TORC1), we will highlight that the molecular organization of enzyme condensates can vary dramatically and that future work aimed at the structural characterization of enzyme condensates will be critical to understand how phase separation regulates enzyme activity and consequently metabolic homeostasis. This information may ultimately facilitate the design of strategies to target the assembly or disassembly of specific enzymes condensates as a therapeutic approach to restore metabolic homeostasis in certain diseases.
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Eoff, R. L., and K. D. Raney. "Helicase-catalysed translocation and strand separation." Biochemical Society Transactions 33, no. 6 (October 26, 2005): 1474–78. http://dx.doi.org/10.1042/bst0331474.

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Helicases are molecular-motor enzymes that manipulate DNA or RNA during replication, repair, recombination, transcription, translation and processing of nucleic acids. The mechanisms for helicase activity have been studied intensely over the past decade. Recent advances in our understanding of the helicase mode of action have led to a general convergence of models that describe this diverse class of enzymes. One mechanism has been proposed that appears to have withstood the test of time, namely the inchworm mechanism. As the name implies, this mechanism involves a process whereby a helicase maintains at least two sites of contact with the nucleic acid. These binding sites can move relative to one another in a sequential fashion, resulting in net movement of the enzyme along the nucleic acid. The inchworm mechanism appears to be applicable to oligomeric states beyond the simple monomeric molecular motor. Although there are certainly many pertinent questions that remain unanswered, striking similarities in both form and function of seemingly disparate enzymes are becoming evident.
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Keçeci, Kübra, Balázs Lemmer, Szabolcs Kertész, Gábor Keszthelyi-Szabó, Zsuzsanna László, and Cecilia Hodúr. "The effect of the implementation of ultrasound in enzyme separation." Analecta Technica Szegedinensia 9, no. 2 (June 12, 2015): 34–41. http://dx.doi.org/10.14232/analecta.2015.2.34-41.

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Enzymes are biological catalysts that generally are designed to do one job well, but to do one job only. Therefore, the enzymes that catalyze the hydrolysis of cellulose to sugar do not break down the sugars. Enzymatic hydrolysis processes have been under development for only 10 years. The important research issues include understanding the processes necessary to render the crystalline cellulose easily digestible, understanding and improving the basic mechanisms in the hydrolysis step, and developing better and less expensive enzymes. The other way to make a process less expensive may be the recycling of enzymes. The essential unit operation in the bioethanol production is the cellulose enzymatic degradation, so the question of recycling is very important. In our work the sonication assisted ultrafiltration was investigated as a potential method for enzyme recycling. The results showed the ultrasound effects the permeate flux since the resistance is reduced by the sonication. The sonicated enzyme keeps its activity so the recycling mechanism might be used for bioethanol production.
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Kazenwadel, F., H. Wagner, B. E. Rapp, and M. Franzreb. "Optimization of enzyme immobilization on magnetic microparticles using 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) as a crosslinking agent." Analytical Methods 7, no. 24 (2015): 10291–98. http://dx.doi.org/10.1039/c5ay02670a.

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Zeitoune, Jessica Florinda. "Enzime recovery by ultrafiltration from broth." Analecta Technica Szegedinensia 8, no. 2 (May 12, 2014): 23–27. http://dx.doi.org/10.14232/analecta.2014.2.23-27.

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Recycling waste products of food industry is important: one side because of the environmental aspects and the other side because of the economic reasons. The one of the most preferred basic material for second generation bio-fuels might be the tobacco (Ábel et al. 2011). The cost of the process depends on the cost of the hydrolysis of cellulose/lignocelluloses i.e. and the cost of the enzymes. These enzymes are very expensive and that is why it is so important to find a good enzyme recovery method. In my research the membrane separation was used for enzyme recovery. Different polyether-sulphone membranes with cut-off value of 7 kDa (PES7) and 10 kDa (PES10) were used for separation the hydrolyzate.
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WANG, DANHUI, ZIYUAN WANG, FEI HE, AMANDA J. KINCHLA, and SAM R. NUGEN. "Enzymatic Digestion for Improved Bacteria Separation from Leafy Green Vegetables." Journal of Food Protection 79, no. 8 (August 1, 2016): 1378–86. http://dx.doi.org/10.4315/0362-028x.jfp-15-581.

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ABSTRACT An effective and rapid method for the separation of bacteria from food matrix remains a bottleneck for rapid bacteria detection for food safety. Bacteria can strongly attach to a food surface or internalize within the matrix, making their isolation extremely difficult. Traditional methods of separating bacteria from food routinely involve stomaching, blending, and shaking. However, these methods may not be efficient at removing all the bacteria from complex matrices. Here, we investigate the benefits of using enzyme digestion followed by immunomagnetic separation to isolate Salmonella from spinach and lettuce. Enzymatic digestion using pectinase and cellulase was able to break down the structure of the leafy green vegetables, resulting in the detachment and release of Salmonella from the leaves. Immunomagnetic separation of Salmonella from the liquefied sample allowed an additional separation step to achieve a more pure sample without leaf debris that may benefit additional downstream applications. We have investigated the optimal combination of pectinase and cellulase for the digestion of spinach and lettuce to improve sample detection yields. The concentrations of enzymes used to digest the leaves were confirmed to have no significant effect on the viability of the inoculated Salmonella. Results reported that the recovery of the Salmonella from the produce after enzyme digestion of the leaves was significantly higher (P <0.05) than traditional sample preparation methods to separate bacteria (stomaching and manually shaking). The results demonstrate the potential for use of enzyme digestion prior to separation can improve the efficiency of bacteria separation and increase the likelihood of detecting pathogens in the final detection assay.
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Sher, Hassan, Hazrat Ali, Muhammad H. Rashid, Fariha Iftikhar, Saif-ur-Rehman, Muhammad S. Nawaz, and Waheed S. Khan. "Enzyme Immobilization on Metal-Organic Framework (MOF): Effects on Thermostability and Function." Protein & Peptide Letters 26, no. 9 (September 16, 2019): 636–47. http://dx.doi.org/10.2174/0929866526666190430120046.

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MOFs are porous materials with adjustable porosity ensuing a tenable surface area and stability. MOFs consist of metal containing joint where organic ligands are linked with coordination bonding rendering a unique architecture favouring the diverse applications in attachment of enzymes, Chemical catalysis, Gases storage and separation, biomedicals. In the past few years immobilization of soluble enzymes on/in MOF has been the topic of interest for scientists working in diverse field. The activity of enzyme, reusability, storage, chemical and thermal stability, affinity with substrate can be greatly improved by immobilizing of enzyme on MOFs. Along with improvement in enzymes properties, the high loading of enzyme is also observed while using MOFs as immobilization support. In this review a detail study of immobilization on/in Metalorganic Frameworks (MOFs) have been described. Furthermore, strategies for the enzyme immobilization on MOFs and resulting in improved catalytic performance of immobilized enzymes have been reported.
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Ullah, Najeeb, Mujaddad Ur Rehman, Abid Sarwar, Muhammad Nadeem, Rubina Nelofer, Hafiz Abdullah Shakir, Muhammad Irfan, et al. "Purification, Characterization, and Application of Alkaline Protease Enzyme from a Locally Isolated Bacillus cereus Strain." Fermentation 8, no. 11 (November 11, 2022): 628. http://dx.doi.org/10.3390/fermentation8110628.

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Among the microbial enzymes protease and amylase are the most valuable enzymes which have been has diversified applications and used extensively because of their capabilities in the degradation of organic wastes, application in biofuels, agricultural, pharmaceuticals, chemical and biotechnological industries. The aim of the current research work was the purification, characterization and application of alkaline proteases extracted from Bacillus cereus AUST-7. Various concentrations of ammonium sulphate were applied for enzyme precipitation. Sephadex-G 100 was used in FPLC system for separation of protease from other proteins. SDS-PAGE was used to measure the molecular weight of required alkaline protease. Relative activities were determined against different pH, temperature, and incubation period to measure the enzymes activity. Stability of pH, temperature and various metal ions and inhibiter were also studied. Purified enzymes were applied on the goat skin to explore the dehairing efficacy. A 6.5 purification fold and 1163.50 U/mg of specific activity were obtained at 70% saturation and 35. 91 purification fold and 8902 U/mg of specific activity were observed after FPLC separation. The 35 kDa molecular size of protease enzyme was exhibited on the SDS-PAGE. The purified enzyme was stable at pH 10, temperature 55 °C and 35 min of incubation period. The purified enzyme was found to be stable at pH 8–11, thermo-stability at 50 °C and phenyl methyl sulphonyl fluoride (PMSF) and di-isopropyl fluorophosphates (DFP) inhibited the enzyme activity. The enzyme has good potential as dehairing agent in leather industries.
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Dissertations / Theses on the topic "Enzymes Separation"

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Haileselassie, Seble Sereke Berhan. "Production of enzyme-modified cheese and bioactive peptides by Lactobacillus and commercial enzymes." Thesis, National Library of Canada = Bibliothèque nationale du Canada, 1999. http://www.collectionscanada.ca/obj/s4/f2/dsk2/ftp03/MQ50782.pdf.

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Perraud, Xavier. "Characterization of lipoxygenases and associated enzymes from selected microorganisms." Thesis, National Library of Canada = Bibliothèque nationale du Canada, 2000. http://www.collectionscanada.ca/obj/s4/f2/dsk1/tape2/PQDD_0032/NQ64642.pdf.

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Wang, Yan. "Pretreatment and Enzymatic Treatment of Spruce : A functional designed wood components separation for a future biorefinery." Doctoral thesis, KTH, Träkemi och massateknologi, 2014. http://urn.kb.se/resolve?urn=urn:nbn:se:kth:diva-150395.

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The three main components of wood, namely, cellulose, hemicellulose, and lignin, can be used in various areas. However, since lignin covalently crosslinks with wood polysaccharides creating networks that is an obstacle for extraction, direct extraction of different wood components in high yield is not an easy matter. One potential approach to overcome such obstacles is to treat the wood with specific enzymes that degrade the networks by specific catalysis. However, the structure of wood is so compact that the penetration of the wood fibers by large enzyme molecules is hindered. Thus, the pretreatment of wood prior to the application of enzymes is necessary, for “opening” the structure. One pretreatment method that was performed in this thesis is based on kraft pulping, which is a well-established and industrialized technique. For untreated wood, the wood fibers cannot be attacked by the enzymes. A relatively mild pretreatment was sufficient for wood polysaccharides hydrolyzed by a culture filtrate. A methanol-alkali mixture extraction was subsequently applied to the samples that were pretreated with two types of hemicellulases, Gamanase and Pulpzyme HC, respectively. The extraction yield increased after enzymatic treatment, and the polymers that were extracted from monocomponent enzyme-treated wood had a higher degree of polymerization. Experiments with in vitro prepared lignin polysaccharide networks suggested that the increased extraction was due to the enzymatic untying. However, the relatively large loss of hemicellulose, particularly including (galacto)glucomannan (GGM), represents a problem with this technique. To improve the carbohydrate yield, sodium borohydride (NaBH4), polysulfide and anthraquinone were used, which increased the yields from 76.6% to 89.6%, 81.3% and 80.0%, respectively, after extended impregnation (EI). The additives also increased the extraction yield from approximately 9 to 12% w/w wood. Gamanase treatment prior to the extraction increased the extraction yield to 14% w/w wood. Sodium dithionite (Na2S2O4) is an alternative reducing agent for the preservation of hemicelluloses because it is less expensive than metal hydrides and only contains sodium and sulfur, which will not introduce new elements to the recovery system. Moreover, Na2S2O4has the potential to be generated from black liquor. Na2S2O4 has some preservation effect on hemicelluloses, and the presence of Na2S2O4 also contributed to delignification. The extraction yield increased to approximately 15% w/w wood. Furthermore, Na2S2O4 has been applied in the kraft pulping process of spruce. The yield and viscosity increased, while the Klason lignin content and kappa number decreased, which represents a beneficial characteristic for kraft pulp. The brightness and tensile strength of the resulting sheets also improved. However, the direct addition of Na2S2O4 to white liquor led to greater reject content. This problem was solved by pre-impregnation with Na2S2O4 and/or mild steam explosion (STEX) prior to the kraft pulping process. Following Na2S2O4 pre-impregnation and mild STEX, the obtained kraft pulp had substantially better properties compared with the properties exhibited after direct addition of Na2S2O4 to the white liquor. The wood structure opening efficiency of mild STEX alone was also tested. The accessibility of the wood structure to enzymes was obtained even at very modest STEX conditions, according to a reducing sugar analysis, and was not observed in untreated wood chips, which were used as a reference. The mechanical effect of STEX appears to be of great importance at lower temperatures, and both chemical and mechanical effects occur at higher STEX temperatures.

QC 20140903

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Rodrigues, Eliana Maria Gonçalves. "Extração liquido-liquido de xilanase por micela reversa numa microcoluna de campanulas pulsadas." [s.n.], 2001. http://repositorio.unicamp.br/jspui/handle/REPOSIP/267501.

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Orientador: Elias Basile Tambourgi
Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia Quimica
Made available in DSpace on 2018-07-28T22:32:01Z (GMT). No. of bitstreams: 1 Rodrigues_ElianaMariaGoncalves_D.pdf: 3569293 bytes, checksum: b7c2f26dc727eabf16b3528d0a1813bb (MD5) Previous issue date: 2001
Resumo: Neste trabalho, foi estudada uma microcoluna agitada por campânulas pulsadas, visando promover um eficiente contato entre as fases através de uma agitação suave, aumentando assim o tempo de contato entre elas no interior da microcoluna e também evitando a desnaturação da enzima. O objetivo deste estudo visou a recuperação da xilanase, produzida pelo fungo Penicillium janthinellum, através da técnica de extração líquido - líquido por micela reversa, para o interior micelar. Para tanto, foi utilizado o agente tensoativo catiônico BDBAC (c1oreto de benzil dodecil bis (hidroxietil) amônio). Foram utilizados planejamentos estatísticos com o intuito de se realizar uma triagem das variáveis significativas no processo: freqüência de pulsação das campânulas, razão entre as fases aquosa/orgânica e condutividade da fase aquosa; a metodologia de superficie de resposta foi empregada para a quantificação dos níveis das mesmas. O trabalho resultou em dois modelos matemáticos, um que representa a recuperação da enzima pura, e outro que representa a recuperação da enzima presente no extrato enzimático bruto. Foram previstos e observados experimentalmente para ambos os modelos obtidos, rendimento em atividade na ordem de 140% para a enzima pura e 43% para o extrato enzimático. Verificou-se neste trabalho que, a metodologia estatística empregada foi de extrema importância e que, a microcoluna estudada tem operação estável e altos rendimentos em atividade foram alcançados
Abstract: In this work, it was studied a microcolumn agitated by puIsed caps, due to promote an efficient contact between the phases in the column and also to avoid the enzyme denaturation and the loss of main proteins properties. This work deals with the purification of xylanase produced by Penicillium janthinellum by liquid-liquid extraction on reverse micelles, to micellar inner, utilizing a cationic surfactant BDBAC (N-benzyI-N-dodecyl-N-bis(2-hydroxyethyl)). It was used design statistical with the intention of realized selection of the main variables in the process: frequency pulsed, volumetric flow and ionic strength; the response surface methodology was employed to the quantified levels of this. This resulted in two major mathematical models: one representing the use of pure enzyme and the other the use of enzymatic extract. It was predict and observed experimentally for both models, obtained activity yield in the order of 140% to the pure enzyme and 43% to the enzymatic extract. It was observed in this work that, the methodology statistical employed was extremely important and that the microcolumn used were stable operation and that high activity yield was reached.
Doutorado
Desenvolvimento de Processos Biotecnologicos
Doutor em Engenharia Química
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Pigeon, Dominique. "Etude des enzymes de synthèse des catécholamines et phosphorylation de la tyrosine hydroxylase de phéochromocytome de rat." Paris 6, 1986. http://www.theses.fr/1986PA066168.

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La structure et le régulation des enzymes de synthèse des catécholamines ont été étudiées dans le phéochromocytome de rat. Nous avons mis au point un protocole permettant la séparation des trois premiers enzymes de la voie de synthèse. Ceci a permis de purifier partiellement la dopamine beta -hydroxylase et de mettre en évidence son inhibiteur endogène, qui a été décrit pour la première fois. Nous avons montré qu'une activité protéine kinase était copurifiée avec la tyrosine hydroxylase, qu'elle phosphoryle. Cette activité a été caractérisé et la séquence primaire du site de phosphorylation a été déterminée.
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Sadler, Andrew Michael. "The separation and immobilisation of a yeast intracellular enzyme." Thesis, University of Surrey, 1988. http://epubs.surrey.ac.uk/847980/.

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The aim of this work was to investigate the problems involved in production and separation of a yeast microsomal enzyme, cytochrome P450. Immobilisation of the microsomal preparation, and utilisation of the immobilised system for the removal of polycyclic aromatic hydrocarbons, particularly the carcinogen benzo(a)pyrene [B(a)P], from aqueous systems was also investigated. High recoveries of the microsomal enzyme were obtained using rapid, low-speed centrifugation (5 minutes, 3000xg) by prior precipitation with a non-ionic polymer, polyethylene glycol (PEG). PEG was found to effectively replace centrifugal acceleration. A semi-empirical mathematical model of the process based on the size distribution of the agglomerates formed and the proportion of the agglomerates sedimented by centrfugation was developed. The effect of PEG was consistent with its increasing the mean effective diameter of protein agglomerates in proportion to PEG concentration to an exponent of 0.619 and with its increasing the spread of the size distribution in proportion to the mean effective agglomerate diameter. Binding spectra studies established that B(a)P binds to the active site of yeast cytochrome P450 and is unaffected by PEG. The B(a)P assay by fluorescence following hexane extraction was also unaffected by PEG. The stability of the microsomal cytochrome P450 preparation at different temperatures was investigated. The enzyme half-life was 66 minutes at 37° and was also unaffected by PEG. The enzyme preparation was satisfactorily immoblilsed by encapsulation in calcium alginate gel and enzyme distribution profiles were determined in sections of alginate stained with coomasie blue by a novel technique using a scanning optical densitometer, Diffusivity coefficients of B(a)P and NADP in alginate gel beads were determined as 7.5 and 2.5 x 10[-10] m[2]/S, respectively by the Tanaka method, fitting solute depletion profiles to Crank's theoretical model. The microsomal enzyme preparation immobilised in alginate beads was used to remove B(a)P from aqueous solution. B(a)P removal was shown to be by a non-specific affinity absorption mechanism and the removal profile was found to correspond well to a theoretical model of a diffusion-reaction system, for an affinity ligand system. The activaiton energy for deactivation of microsomal P450 was measured as 33.56kJ/mole.
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Olceroglu, Ayse Hande. "Chiral Separations By Enzyme Enhanced Ultrafiltration: Fractionation Of Racemic Benzoin." Master's thesis, METU, 2006. http://etd.lib.metu.edu.tr/upload/12607460/index.pdf.

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In this study, a methodology for separation of chiral molecules, by using enhanced ultrafiltration system was developed. Benzoin was the model chiral molecule studied. In the scope of developing this methodology, some parameters were investigated in the preliminary ultrafiltration experiments in order to set the operation conditions for enhanced ultrafiltration experiments. Due to the slight solubility of benzoin in pure water, 15% (v/v) Polyethylene glycol (PEG 400) and 30 % (v/v) Dimethyl sulfoxide (DMSO) were selected as cosolvents. Because of the high retention capacity of RC-10000 Da membranes for benzoin, a membrane saturation strategy was developed. In polymer enhanced ultrafiltration (PEUF) experiments bovine serum albumin (BSA) was used as ligand. Effects of ligand concentration and pH on total benzoin retention and on enantiomeric excess (ee %) were investigated. Benzoin concentration was almost kept constant at ~10 ppm and ~50 ppm for 15% (v/v) PEG 400 and 30 % (v/v) DMSO cosolvents, respectively. It was observed that the increase either in pH or in BSA concentration yielded an increase in total benzoin retention. In 15% (v/v) PEG 400-water, with BSA concentration of 10000 ppm, at pH 10, total benzoin retention reached to 48.7%. For this cosolvent, at different pH values and at different BSA concentrations, all ee % values were about or less than 10%. When 50000 ppm BSA was dissolved in 30 % (v/v) DMSO-water, total benzoin retention increased to 41.3% at pH 10 and ee % reached 16.7 % at pH 11. In enzyme enhanced ultrafiltration (EEUF) experiments, specific to benzoin, apo form of Benzaldehyde Lyase (BAL, E.C. 4.1.2.38) was used as ligand. These experiments were performed with constant ~ 10 ppm benzoin concentration in only 15% (v/v) PEG 400 &ndash
water solvent. Effect of BAL concentration on total benzoin retention and ee% was investigated. It was found that
for all the studied BAL concentrations in the range of 650- 1936 ppm total benzoin retention and ee % were kept almost constant at ~75% and ~60%, respectively.
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Prinz, Axel [Verfasser]. "Enzyme Separation Using Aqueous Two-Phase Extraction: Experiment, Model and Simulation / Axel Prinz." München : Verlag Dr. Hut, 2014. http://d-nb.info/1053859686/34.

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Chen, Cheau-Yun. "Studies of Enzyme Mechanism Using Isotopic Probes." Thesis, North Texas State University, 1987. https://digital.library.unt.edu/ark:/67531/metadc331996/.

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The isotope partitioning studies of the Ascaris suum NAD-malic enzyme reaction were examined with five transitory complexes including E:NAD, E:NAD:Mg, E:malate, E:Mg:malate, and E:NAD:malate. Three productive complexes, E:NAD, E:NAD:Mg, and E:Mg:malate, were obtained, suggesting a steady-state random mechanism. Data for trapping with E:14C-NAD indicate a rapid equilibrium addition of Mg2+ prior to the addition of malate. Trapping with 14C-malate could only be obtained from the E:Mg2+:14C-malate complex, while no trapping from E:14C-malate was obtained under feasible experimental conditions. Most likely, E:malate is non-productive, as has been suggested from the kinetic analysis. The experiment with E:NAD:malate could not be carried out due to the turnover of trace amounts of malate dehydrogenase in the pulse solution. The equations for the isotope partitioning studies varying two substrates in the chase solution in an ordered terreactant reaction were derived, allowing a determination of the relative rates of substrate dissociation to the catalytic reaction for each of the productive transitory complexes. NAD and malate are released from the central complex at an identical rate, equal to the catalytic rate.
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Chen, Zhiqiang. "NANOMETER-SCALE MEMBRANE ELECTRODE SYSTEMS FOR ACTIVE PROTEIN SEPARATION, ENZYME IMMOBILIZATION AND CELLULAR ELECTROPORATION." UKnowledge, 2014. http://uknowledge.uky.edu/cme_etds/33.

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Automated and continuous processes are the future trends in downstream protein purification. A functionalized nanometer-scale membrane electrode system, mimicking the function of cell wall transporters, can selectively capture genetically modified proteins and subsequently pump them through the system under programmed voltage pulses. Numerical study of the two-step pulse pumping cycles coupled with experimental His-GFP releasing study reveals the optimal 14s/1s pumping/repel pulse pumping condition at 10 mM bulk imidazole concentration in the permeate side. A separation factor for GFP: BSA of 9.7 was achieved with observed GFP electrophoretic mobility of 3.1×10-6 cm2 s-1 V-1 at 10 mM bulk imidazole concentration and 14 s/1 s pumping/repel duration. The purification of His6-OleD Loki variant directly from crude E. coli extracts expression broth was demonstrated using the pulse pumping process, simplifying the separation process as well as reducing biopharmaceutical production costs. The enzymatic reactions showed that His6-OleD Loki was still active after purification. A nanoporous membrane/electrode system with directed flow carrying reagents to sequentially attached enzymes to mimic nature’s enzymes-complex system was demonstrated. The substrates residence time on the immobilized enzyme can be precisely controlled by changing the pumping rate and thereby prevent a secondary hydrolysis reaction. Immobilized enzyme showed long term storage longevity with activity half-life of 50 days at 4℃ and the ability to be regenerated. One-step immobilization and purification of His-tagged OleD Loki variant directly from expression broth, yielded 98% Uridine Diphosphate glycosylation and 80% 4-methylumbelliferone glycosylation conversion efficiency for the sequential reaction. A flow-through electroporation system, based on a novel membrane/electrode design, for the delivery of membrane-impermeant molecules into Model Leukocyte cells was demonstrated. The ability to apply low voltage between two short distance electrodes contributes to high cell viability. The flow-through system can be easily scaled-up by varying the micro-fluidic channel geometry and/or the applied voltage pulse frequency. More importantly, the system allows the electrophoretical pumping of molecules from the reservoir across the membrane/electrode system to the micro-fluidic channel for transfection, which reduces large amount of reagents used.
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Books on the topic "Enzymes Separation"

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Electrophoresis of enzymes: Laboratory methods. Berlin: Springer-Verlag, 1994.

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Nath, Gupta Munishwar, ed. Methods for affinity-based separations of enzymes and proteins. Basel: Birkhäuser Verlag, 2002.

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1939-, Karger Barry L., and Hancock William S, eds. High resolution separation and analysis of biological macromolecules. San Diego: Academic Press, 1996.

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Gupta, Munishwar Nath, ed. Methods for Affinity-Based Separations of Enzymes and Proteins. Basel: Birkhäuser Basel, 2002. http://dx.doi.org/10.1007/978-3-0348-8127-2.

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Acquaah, George. Practical protein electrophoresis for genetic research. Portland, Or: Dioscorides Press, 1992.

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(Editor), John N. Abelson, Melvin I. Simon (Editor), and Helmut Sies (Editor), eds. Carotenoids, Part A, Chemistry, Separation, Quantitation, and Antioxidation, Volume 213 (Methods in Enzymology). Academic Press, 1992.

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(Editor), John N. Abelson, Melvin I. Simon (Editor), and Helmut Sies (Editor), eds. Carotenoids, Part A, Chemistry, Separation, Quantitation, and Antioxidation, Volume 213 (Methods in Enzymology). Academic Press, 1992.

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(Editor), John N. Abelson, Melvin I. Simon (Editor), Barry L. Karger (Editor), and William Hancock (Editor), eds. Methods in High Resolution Separation and Analysis of Biological Macromolecules : Applications (Methods in Enzymology Series, Vol 271, Part B) (Methods in Enzymology). Academic Press, 1996.

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(Editor), John N. Abelson, Melvin I. Simon (Editor), Harry Walter (Editor), and Göte Johansson (Editor), eds. Aqueous Two-Phase Systems (Methods in Enzymology). Academic Press, 1994.

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Abelson, John N., Harry Walter, Melvin I. Simon, and Göte Johansson. Aqueous Two-Phase Systems. Elsevier Science & Technology Books, 1994.

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Book chapters on the topic "Enzymes Separation"

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Rothe, Gunter M. "A Compilation of Protocols to Visualize Enzymes Following Electrophoretic Separation." In Electrophoresis of Enzymes, 181–271. Berlin, Heidelberg: Springer Berlin Heidelberg, 1994. http://dx.doi.org/10.1007/978-3-642-79069-0_6.

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Humphrey, Cara E., Marwa Ahmed, Ashraf Ghanem, and Nicholas J. Turner. "Application of Enzymes in Kinetic Resolutions, Dynamic Kinetic Resolutions and Deracemization Reactions." In Separation of Enantiomers, 123–60. Weinheim, Germany: Wiley-VCH Verlag GmbH & Co. KGaA, 2014. http://dx.doi.org/10.1002/9783527650880.ch4.

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Gupta, Munishwar N., and Ipsita Roy. "Affinity-Based Separation: An Overview." In Methods for Affinity-Based Separations of Enzymes and Proteins, 1–15. Basel: Birkhäuser Basel, 2002. http://dx.doi.org/10.1007/978-3-0348-8127-2_1.

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Wesselingh, J. A. "Large Scale Separation of Intracellular Enzymes (how not to do it)." In Recent Advances in Biotechnology, 69–88. Dordrecht: Springer Netherlands, 1992. http://dx.doi.org/10.1007/978-94-011-2468-3_4.

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Persson, Lars-Olof, and Göte Johansson. "Separation of Calvin Cycle Enzymes, from Spinach Chloroplasts, by Affinity Partitioning with Triazine Dye Ligands." In Progress in Photosynthesis Research, 495–98. Dordrecht: Springer Netherlands, 1987. http://dx.doi.org/10.1007/978-94-017-0516-5_105.

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Deshpande, S. S. "Separation and Solid-Phase Systems." In Enzyme Immunoassays, 193–228. Boston, MA: Springer US, 1996. http://dx.doi.org/10.1007/978-1-4613-1169-0_7.

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Johansson, Göte. "Affinity Partitioning of Enzymes." In Separations Using Aqueous Phase Systems, 7–14. Boston, MA: Springer US, 1989. http://dx.doi.org/10.1007/978-1-4684-5667-7_2.

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Iorio, G., V. Calabro’, and S. Todisco. "Enzyme Membrane Reactors." In Membrane Processes in Separation and Purification, 149–67. Dordrecht: Springer Netherlands, 1994. http://dx.doi.org/10.1007/978-94-015-8340-4_8.

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Rothe, Gunter M. "Methods for Separating Native Enzymes." In Electrophoresis of Enzymes, 71–125. Berlin, Heidelberg: Springer Berlin Heidelberg, 1994. http://dx.doi.org/10.1007/978-3-642-79069-0_3.

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Jaklitsch, Anna. "Separation-Free Enzyme Immunoassay for Haptens." In Enzyme-Mediated Immunoassay, 33–55. Boston, MA: Springer US, 1985. http://dx.doi.org/10.1007/978-1-4684-5012-5_3.

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Conference papers on the topic "Enzymes Separation"

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Menasni, S., W. Hornebeck, L. Robert, and Y. Legrand. "ELASTASE TYPE ACTIVITY OF ENDOTHELIAL CELLS." In XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1643360.

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Elastin degrading enzymes have been reported in the vessel wall and both fibroblasts and smooth muscle cells have been shown to produce elastase type enzymes in culture. Data is presented here showing that porcine aortic endothelial cells produce enzyme activities hydrolyzing elastin and synthetic substrates I Sue Ala Ala Ala nitroanilide, SAPNAI considered specific for elastase. Enzyme activity against the SAPNA but not against H-elastin was found to be associated with the cells after triton lysis .This activity was not secreted into the culture medium . The elastolytic activity has been partially characterized in relation to the kinetic of hydrolysis, pH optimum and susceptibility to different inhibitors. These studies revealed the presence of at least two enzymes: a metalo-protease with a pH optimum of 7.5 which accounts for approx. 80% of the total activity, and a serine protease with pH optimum of 8.0 which accounts for the remaining 20% . When the conditioned culture medium was studied, virtually no proteolytic activity could be detected even after activation with an organomercurial agent. However fractionation of the culture medium by gel filtration on HPLC resulted in elastolytic activity both against H-elastin and SAPNA. Proteolytic activity against casein could also be revealed after separation on SDS-PAGE. It is likely that these separation techniques remove an inhibitor also produced by the endothelial cells and allow the expression of proteolytic activity. That the elastolytic activity and the caseinolytic activity revealed by HPLC and PAGE respectively represent the activity of the same enzyme hase not yet been determined, and its relationship to the Stromelysin described by Herron et al(J. Biol. Chem. , 1986, 261. 2810-2813) in rabbit brain capillary endothelial cells is being investigated.
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Wyatt, Karla E. K., Jonathan W. Bourne, and Peter A. Torzilli. "Deformation-Dependent Enzyme Cleavage of Collagen." In ASME 2007 Summer Bioengineering Conference. American Society of Mechanical Engineers, 2007. http://dx.doi.org/10.1115/sbc2007-176502.

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Collagen degradation is a mechanism for normal musculoskeletal development and extracellular matrix (ECM) maintenance, and in response to trauma, disease and inflammation. Matrix metalloproteinases (MMP-1, 8, and 13, the collagenases) are the primary enzymes that act to degrade collagen. These MMPs gain access to the collagen triple helix by binding to the enzyme’s attachment domain along the α-chains, followed by separation (unwinding) of the α-chains to expose the 3/4–1/4 cleavage site, and then cleavage of the α-chain by the enzyme’s catalytic domain [3, 5].
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Kuyas, C., A. Haeberli, and P. W. Straub. "SEPARATION OF FIBRINOGEN FRAGMENTS ON GLYPRQARGPROLYS-FRACTOGEL." In XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1642885.

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The tetrapeptide GlyProArgPro, which corresponds to the newly exposed N-terminal sequence of fibrin a-polypeptide chain after the action of thraribin, has a binding site in the C-terminal part of the ;-chain as suggested by several authors. Using Gly-ProArgProLys-Fractogel (GPRPK) chrcmatography we tried to isolate a fibrinogen fragment, obtained with different enzymes and conditions, which includes the binding site for GPRP. Human fibrinogen was digested by plasmin in presence and absence of Ca-ions, and the resulting lysates were applied in 0.05 M triethanolamine (TEA), 0.1 M NaCl pH 7.4 to the GPRPK-Fractogel. The gel was washed extensively with TEA-buffer and the adsorbed protein was eluted with 6M urea in TEA-buffer. The protein containing fractions were analyzed iiununologically with anti-fragment E- and with anti-fragment D antibodies. Human fibrinogen was also digested with endopeptidase Arg-C in O.IM NaHco3 pH 8.0 at 37C over night. The enzyme cleaves fibrinogen to fragments, one of which comprising the y-chain sequence 275-375 which is said to contain the fibrin polymerization site. The Arg C-lysate was chromatographed on GPRPK-Fractogel. All fragments were analyzed by SDS-electrophoresis and by reversed-phase HPLC.Fragment D1 was the only fibrinogen fragment which was adsorbed on GPRPK-Fractogel. All other assayed fibrinogen fragments obtained by enzymatic cleavage, showed no affinity to GPRPK-Fractogel. These results demonstrate that for the binding of GPRP to fibrinogen a conformationally intact γ-chain remnant of the fragment D is required.One step chrcmatography using GPRPK-Fractogel can thus also be used to isolate fragment D1 in high purity fran plasmin lysates of fibrinogen.
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Bogojevic, Oliver, Carl Arevang, and Zheng Guo. "Synthesis of complex phospholipid species." In 2022 AOCS Annual Meeting & Expo. American Oil Chemists' Society (AOCS), 2022. http://dx.doi.org/10.21748/rlyh7861.

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Phospholipids are essential for the preservation of life on the planet and carry numerous critical roles and functions, including being the main constituents of the cell-membranes in both eukaryotic and prokaryotic cells, providing (more bioavailable) energy, and maintaining chemical and electrical processes in the body. The structural characteristics of phospholipids can vary greatly among species, however, commonly consist of a hydrophilic region (phosphate-containing head-group) and a hydrophobic region (fatty acids, €œtails€), providing the amphiphilic features and unique functions. The countless number of possible configurations enables the continuous synthesis of novel phospholipid species. The synthesis of specific phospholipids, so-called €œdesigner-phospholipids€, is commonly carried out through modifications of more common and easily accessible phospholipid species, catalyzed by the use of either non-specific chemical catalysts or specific enzymes. Enzymatic methods, being most prominent, are often using biphasic reaction systems, allowing for the easy reuse of enzymes and separation of polar compounds, offering more environmentally friendly approaches'. The synthesis of complex phospholipids such as cardiolipins (CLs) and bis(mono/di-acylglycero)phopshates (BMPs/BDPs) have significant value as they carry the unique ability to contain multiple fatty acids, which in turn can be linked to a range of positive health effects. The positive health effects of fish oils (EPA/DHA) are today a hot topic, which in combination with complex phospholipids present great potential for future applications. Additionally, new phospholipid species are continuously under development utilizing completely new synthetic systems with environmentally friendly approaches' in focus. Modern methods centralized on the combinatorial use of ionic liquids and enzymes for the production of novel phospholipids species reduce the use of organic solvents, allowing for the incorporation of fatty acid esters of hydroxy fatty acids (FAHFAs) into phospholipids. The science behind the synthesis of phospholipids is continuously developing for an increased amount of different applications.
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Trachtenberg, Michael C., Martin L. McGregor, Chingkuang Tu, Philip J. Laipis, Richard C. Willson, John F. Kennedy, Marion Paterson, and Frederick B. Rudolph. "Enzyme-Enhanced Membranes for Gas Separation." In International Conference On Environmental Systems. 400 Commonwealth Drive, Warrendale, PA, United States: SAE International, 1999. http://dx.doi.org/10.4271/1999-01-1961.

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Jordan, R. E., J. Kilpatrick, J. Nelson, J. O. New gren, and M. A. Fournel. "HEPARIN DIRECTS THE INACTIVATION OF ANTITHROMBIN BY ELASTASE." In XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1643770.

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In apparent contradiction to its anticoagulant activity, we have observed a previously undetected, and potentially opposing function for heparin: a distinct heparin-dependency for the in vitro inactivation of highly-purified human antithrombin by neutrophil elastase. Similar to its ability to accelerate antithrombin-mediated inhibition of coagulation enzymes, anticoagulantly-active heparin was also found to stimulate the rate of inactivation of antithrombin by the neutrophil enzyme.In the absence of heparin, or in the presence of the heparin antagonists platelet factor 4 or polybrene, little or no inactivation of antithrombin occurred. Catalytic amounts of heparin and elastase caused the complete inactivation of antithrombin (approximate molar ratio of 1:1:400 respectively) in 5-10 minutes. The loss of heparin binding affinity by the elastase-cleaved form of antithrombin permitted its separation from active antithrombin by heparin-agarose chromatography.The purified elastase-inactivated antithrombin was injected into rabbits for determination of its comparative clearance behavior. In contrast to intact, functional antithrombin (t 1/2 >30 hours) and the thrombin-antithrombin (T-AT) complex (t 1/2 previously shown to be minutes), elastase-inactivated antithrombin circulated for approximately 13 hours. This prolonged clearance relative to the T-AT complex may suggest an alternative explanation for the circulating, non-functional antithrombin observed in certain coagulopathic states. In summary, these results point to a potential and unexpected role for heparin in directing the inactivation of antithrombin and suggest a possible in vivo mechanism for neutralizing the usually non-thrombogenic nature of the vascular lining.
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Udoh, Tinuola, and Osadebamen Aigbodion. "Investigation on Effect of Enzyme on Oil-Brine Emulsification." In SPE Nigeria Annual International Conference and Exhibition. SPE, 2022. http://dx.doi.org/10.2118/211906-ms.

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Abstract In this paper, the capacity of enzyme to influence brine-in-oil and oil-in-brine emulsions was investigated. The emulsion stability index method was used to monitor the effect of varied enzyme concentrations (1-, 5- and 10 wt.%) on oil-brine emulsion stability and separation process. The result of the study shows that the addition of different concentrations of enzyme to oil-brine mixtures enhanced the mixing and separation of the emulsions at varied capacities. Faster oil-brine separation was observed with increase in enzyme concentrations, but better mixing and higher emulsion stability was observed with lower concentration of enzyme. The result of this study is of a great significance to enzyme enhanced oil recovery application process in which good oil-brine mixture is require for the recovery of the residual oil saturation from the reservoir rock pores and the separation of oil and brine that is required after production at the surface.
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Gray, E., P. J. Kerry, S. J. Edwards, and T. W. Barrowcliffe. "PROCOAGULANT ACTIVITY OF PLATELET LIPCKYGENASE PRODUCTS." In XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1643945.

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Arachidomc acid is metabolised by cyclo-oxygenase and lipoxygenase enzymes in platelets. Cyclo-oxygenase produces the highly pro-aggregatory thromboxane A2 but the physiological significance of the lipoxygenase pathway in platelets remains uncertain. Arachidonic acid can also be converted to biologically active peroxides by free radical induced autoxidation, and previous studies have shown that such products generate large amounts of thrombin in platelet-free plasma, via their interaction with plasma lipoproteins.In the present study, we have investigated the procoagulant activities of platelet lipoxygenase products and compared them with autoxidised arachidonic acid. Platelet concentrates were incubated with arachidonic acid and indomethacin for 30 minutes and the products extracted with ethyl acetate. After drying down, reconstitution in ethanol and partitioning with petrolehm ether to remove unchanged acid, contaminating platelet phospholipid was removed by TLC. By inclusion of 14C-arachidonic acid, average conversion was estimated as 17% in six experiments.The products promoted the generation of large amounts of thrombin in platelet-free plasma; the peak thrombin averaged 23.1 iu/ml with three different batches. The activity was similar to that of a procoagulant phospholipid (PL) but, unlike PL, the lipoxygenase products did not shorten the kaolin recalcification time, confirming the absence of platelet PL contamination, and were virtually inactive in lipoprotein-free plasma. The activity of the lipoxygenase products was therefore similar to that of autoxidised arachidonic acid in its requirement for plasma lipoproteins. Further TLC separation of the products showed that the activity was associated with a minor component of the mixture which was active at plasma concentrations below 10 μg/ml.These results suggest a possible role for platelet lipoxygenase products in the coagulation system and provide a novel link between platelets, lipoproteins and coagulation which could be inportant in the pathogenesis of atherosclerosis.
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Ju, Lu-Kwang, Abdullah Al Loman, Md Fauzul Kabir, Qian Li, and S. M. Mahfuzul Islam. "Enzyme-based soy processing." In 2022 AOCS Annual Meeting & Expo. American Oil Chemists' Society (AOCS), 2022. http://dx.doi.org/10.21748/hrib7435.

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Soybeans contain three major components: protein (ca. 40%), carbohydrate (25–30%) and oil (18–20%). To maximize value and minimize waste in soy processing, all these components should be collected and utilized. Current processing was originally designed to maximize oil extraction. It tends to make protein and carbohydrate separation from the remaining meal more difficult and, as a result, can limit their uses and reduce their value. We have been developing an enzyme-based processing method which, in a single step, enables the recovery of oil, protein, and sugar in separate streams. Further, oil and protein present in soybeans as individually “packaged” oil bodies (oleosomes) and protein bodies. This solvent-free, enzyme-based processing allows collection of intact oleosomes and protein bodies without alteration by heat, solvent, or mechanical pressing. This new processing method can maximize the recovery of nutritional and industrial/economic value of all major soybean components. We also develop and optimize the production of enzyme particularly suitable for soy processing.
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Oncescu, Vlad, and David Erickson. "A Microfabricated Enzyme-Free Glucose Fuel Cell for Implantable Devices." In ASME 2011 International Mechanical Engineering Congress and Exposition. ASMEDC, 2011. http://dx.doi.org/10.1115/imece2011-62893.

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In the past decade the scientific community has showed considerable interest in the development of implantable medical devices. Such devices have low power requirements and can potentially be operated through fuel cells using reactants present in the body such as glucose and oxygen instead of non-rechargeable lithium batteries. In this paper we present a thin, enzyme-free fuel cell with high current density and good stability at a current density of 10μA cm−2. The fuel cell uses a stacked electrode design in order to achieve glucose and oxygen separation. In addition, it uses a porous carbon paper support for the anodic catalyst layer which reduces the amount of platinum or other noble metal catalysts required for fabricating high surface area electrodes with good reactivity. The peak power output of the fuel cell is approximately 2μW cm−2 and has a sustainable power density of 1.5μW cm−2 at 10μA cm−2. An analysis on the effects of electrode thickness and inter electrode gap on the maximum power output of the fuel cell is also performed.
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Reports on the topic "Enzymes Separation"

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Hawes, M. C. Role of pectolytic enzymes in the programmed separation of cells from the root cap of higher plants. Final report. Office of Scientific and Technical Information (OSTI), March 1995. http://dx.doi.org/10.2172/41268.

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Carpita, Nicholas C., Ruth Ben-Arie, and Amnon Lers. Pectin Cross-Linking Dynamics and Wall Softening during Fruit Ripening. United States Department of Agriculture, July 2002. http://dx.doi.org/10.32747/2002.7585197.bard.

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Our study was designed to elucidate the chemical determinants of pectin cross-linking in developing fruits of apple and peach and to evaluate the role of breakage cross-linkages in swelling, softening, and cell separation during the ripening. Peaches cell walls soften and swell considerably during the ripening, whereas apples fruit cells maintain wall firmness but cells separate during late stages of ripening. We used a "double-reduction" technique to show that levels of non-methyl esters of polyuronic acid molecules were constant during the development and ripening and decreased only in overripe fruit. In peach, methyl and non-methyl esters increased during the development and decreased markedly during the ripening. Non-methyl ester linkages in both fruit decreased accompanied fruit softening. The identity of the second component of the linkage and its definitive role in the fruit softening remain elusive. In preliminary examination of isolated apples cell walls, we found that phenolic compounds accumulate early in wall development but decrease markedly during ripening. Quantitative texture analysis was used to correlate with changes to wall chemistry from the fresh-picked ripe stage to the stage during storage when the cell separation occurs. Cell wall composition is similar in all cultivars, with arabinose as the principal neutral sugar. Extensive de-branching of these highly branched arabinans pre-stages softening and cell-cell separation during over-ripening of apple. The longer 5-arabinans remain attached to the major pectic polymer rhamnogalacturonan I (RG I) backbone. The degree of RG I branching, as judged from the ratios of 2-Rha:2,4-Rha, also decreases, specially after an extensive arabinan de-branching. Loss of the 4-Rham linkages correlated strongly with the softening of the fruit. Loss of the monomer or polymer linked to the RG I produce directly or indirectly the softening of the fruit. This result will help to understand the fruit softening and to have better control of the textural changes in fruit during the ripening and especially during the storage. 'Wooliness', an undesirable mealy texture that is induced during chilling of some peach cultivars, greatly reduces the fruit storage possibilities. In order to examine the hypothesis that the basis for this disorder is related to abnormality in the cell wall softening process we have carried out a comparative analysis using the resistant cultivar, Sunsnow, and a sensitive one, Hermosa. We investigated the activity of several pectin- and glycan-modifying enzymes and the expression of their genes during ripening, chilling, and subsequent shelf-life. The changes in carbohydrate status and in methyl vs. non-methyl uronate ester levels in the walls of these cultivars were examined as well to provide a basis for comparison of the relevant gene expression that may impact appearance of the wooly character. The activities of the specific polygalacturonase (PGase) and a CMC-cellulase activities are significantly elevated in walls of peaches that have become wooly. Cellulase activities correlated well with increased level of the transcript, but differential expression of PGase did not correspond with the observed pattern of mRNA accumulation. When expression of ethylene biosynthesis related genes was followed no significant differences in ACC synthase gene expression was observed in the wooly fruit while the normal activation of the ACC oxidase was partially repressed in the Hermosa wooly fruits. Normal ripening-related loss of the uronic acid-rich polymers was stalled in the wooly Hermosa inconsistent with the observed elevation in a specific PGase activity but consistent with PG gene expression. In general, analysis of the level of total esterification, degree of methyl esterification and level of non-methyl esters did not reveal any major alterations between the different fruit varieties or between normal and abnormal ripening. Some decrease in the level of uronic acids methyl esterification was observed for both Hermosa and Sunsnow undergoing ripening following storage at low temperature but not in fruits ripening after harvest. Our results support a role for imbalanced cell wall degradation as a basis for the chilling disorder. While these results do not support a role for the imbalance between PG and pectin methyl esterase (PME) activities as the basis for the disorder they suggest a possible role for imbalance between cellulose and other cell wall polymer degradation during the softening process.
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Shahak, Yosepha, and Donald R. Ort. Physiological Bases for Impaired Photosynthetic Performance of Chilling-Sensitive Fruit Trees. United States Department of Agriculture, May 2001. http://dx.doi.org/10.32747/2001.7575278.bard.

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Chilling-sensitivity is an important agricultural problem in both the U.S. and Israel. Most research attention has focused so far on herbaceous crop plants, even though the problem is also acute in the fruit tree industry. Under BARD funding we made substantial progress in identifying the mechanisms involved in the disruption of photosynthesis following a chill in mango. Our investigation with fruit trees has been substantially accelerated by drawing on our knowledge and experience with herbaceous crops. The four original research objectives, focused or discovering the underlying mechanisms of chill-induced inhibition of photosynthesis in fruit trees, and the main achievements are listed below. [1] Separating stomatal from non-stomatal components of chilling on photosynthesis in fruit trees. We found evidence that the dark chill-induced inhibition of photosynthesis in mango was E combination of both stomatal and mesophyll components. [2] Differentiating photo damage from light-induced photo protection of photosystem II (PSII). Dark chilling exacerbate high light photoinhibition, as a result of primary inhibition in the carbor reduction cycle. Nevertheless, in Israeli orchards we observed chronic photoinhibition of PSII photochemistry in the winter. This photo damage was reversible over a few days if sunlight was attenuated with filters or night temperature rose. Practical implications of this finding deserve further investment. Additional achievement was the development of a new biophysical tool to study macro-structural changes of LHCII particles in intact, attached leaves. [3] Determine the role of oxidative stress in the dark-chilling-induced inhibition, with emphasis on oxygen radical scavenging, lipid peroxidation and redox-controlled carbon-cycle enzymes. We found an increase in lipid peroxidation following a dark chill, and partial protective effects or an antioxidant. However, the photoinhibition observed in mango orchards in Israel during the winter did not appear to be a general oxidative stress. [4] Investigate whether chilling interferes with the diurnal and circadian rhythm of gene expression of key photosynthetic proteins as has been shown for chilling-sensitive crop plants. The results indicated that most of the circadian rhythm in photosynthesis was due to reduced lea: internal CO2 concentrations during the subjective night, as a result of rhythmic stomatal closure Chilling-induced interference with circadian timing in mango, does not play the central role in chilling inhibition of photosynthesis that has previously been demonstrated in certain chilling sensitive herbaceous plants. Practical implications of the research achievements are feasible, but require few more years of research.
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