- Tytuł:
- Chitinolytic enzymes from bacterium inhabiting human gastrointestinal tract - critical parameters of protein isolation from anaerobic culture
- Autorzy:
-
Dušková, Jarmila
Tishchenko, Galina
Ponomareva, Evgenia
Šimůnek, Jiří
Koppová, Ingrid
Skálová, Tereza
Štěpánková, Andrea
Hašek, Jindřich
Dohnálek, Jan - Powiązania:
- https://bibliotekanauki.pl/articles/1039931.pdf
- Data publikacji:
- 2011
- Wydawca:
- Polskie Towarzystwo Biochemiczne
- Tematy:
-
chitinolytic enzymes
anaerobic cultivation
protein isolation - Opis:
- The object of this study are chitinolytic enzymes produced by bacterium Clostridium paraputrificum J4 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 showing specific activities of endo-, exo-chitinase and N-acetyl-β-glucosaminidase 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.
- Źródło:
-
Acta Biochimica Polonica; 2011, 58, 2; 261-263
0001-527X - Pojawia się w:
- Acta Biochimica Polonica
- Dostawca treści:
- Biblioteka Nauki
Artykuł