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Chitinolytic enzymes from bacterium inhabiting human gastrointestinal tract - critical parameters of protein isolation from anaerobic culture
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SYSNO ASEP 0363003 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Chitinolytic enzymes from bacterium inhabiting human gastrointestinal tract - critical parameters of protein isolation from anaerobic culture Author(s) Dušková, Jarmila (UMCH-V) RID
Tishchenko, Galina (UMCH-V) RID
Ponomareva, E. (RU)
Šimůnek, Jiří (UZFG-Y) RID
Koppová, Ingrid (UZFG-Y) RID
Skálová, Tereza (UMCH-V) RID
Štěpánková, Andrea (UMCH-V)
Hašek, Jindřich (UMCH-V) RID
Dohnálek, Jan (UMCH-V) RIDSource Title Acta Biochimica Polonica - ISSN 0001-527X
Roč. 58, č. 2 (2011), s. 261-263Number of pages 3 s. Language eng - English Country PL - Poland Keywords chitinolytic enzymes ; anaerobic cultivation ; protein isolation Subject RIV EE - Microbiology, Virology R&D Projects GA310/09/1407 GA ČR - Czech Science Foundation (CSF) GA305/07/1073 GA ČR - Czech Science Foundation (CSF) CEZ AV0Z40500505 - UMCH-V (2005-2011) AV0Z50450515 - UZFG-Y (2005-2011) UT WOS 000292350900018 Annotation The object of this study are chitinolytic enzymes produced by bacterium Clostridium paraputrificum 14 isolated from the gastrointestinal tract of a healthy human. In particular, we focus on the development of purification protocols, determination of properties of the enzymes and their activity profiles. The process of bacteria cultivation and isolation of chitinolytic complex of enzymes was optimized. A range of various purification procedures were used such as ultrafiltration, precipitation, chromatographic separations (ion-exchange, size exclusion, chromatofocusing) in altered combinations. The optimal purification protocol comprises two or three steps. Individual samples were analyzed by SDS/PAGE electrophoresis and after renaturation their activity could be detected using zymograms. Mass spectroscopy peptide fragment analysis and MALDI analysis of the purest samples indicate presence of endochitinase B (molecular mass about 85 kDa) and of 60-kDa endo- and exochitinases. Workplace Institute of Macromolecular Chemistry Contact Eva Čechová, cechova@imc.cas.cz ; Tel.: 296 809 358 Year of Publishing 2012 Electronic address http://www.actabp.pl/pdf/2_2011/261.pdf
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