Počet záznamů: 1
Chitinolytic enzymes from bacterium inhabiting human gastrointestinal tract - critical parameters of protein isolation from anaerobic culture
0363003 - UMCH-V 2012 RIV PL eng J - Článek v odborném periodiku
Dušková, Jarmila - Tishchenko, Galina - Ponomareva, E. - Šimůnek, Jiří - Koppová, Ingrid - Skálová, Tereza - Štěpánková, Andrea - Hašek, Jindřich - Dohnálek, Jan
Chitinolytic enzymes from bacterium inhabiting human gastrointestinal tract - critical parameters of protein isolation from anaerobic culture.
Acta Biochimica Polonica. Roč. 58, č. 2 (2011), s. 261-263 ISSN 0001-527X
Grant CEP: GA ČR GA310/09/1407; GA ČR GA305/07/1073
Výzkumný záměr: CEZ:AV0Z40500505; CEZ:AV0Z50450515
Klíčová slova: chitinolytic enzymes * anaerobic cultivation * protein isolation
Kód oboru RIV: EE - Mikrobiologie, virologie
Impakt faktor: 1.491, rok: 2011
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.
Trvalý link: http://hdl.handle.net/11104/0199145