Chitinolytic enzymes from bacterium inhabiting human gastrointestinal tract - critical parameters of protein isolation from anaerobic culture

• Autorzy: Duskova J. , Tishchenko G. , Ponomareva E. , Simunek J. , Koppova I. , Skalova T. , Stepankova A. , Hasek J. , Dohnalek J.
• Języki publikacji: EN

Abstrakty

EN

 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.

• Wydawca: -
• Czasopismo: Acta Biochimica Polonica
• Rocznik: 2011
• Tom: 58
• Numer: 2
• Opis fizyczny: p.261-163,fig.,ref.

Twórcy

autor

Duskova J.

• Institute of Macromolecular Chemistry, Academy of Sciences of the Czech Republic, Praha, Czech Republic

autor

Tishchenko G.

autor

Ponomareva E.

autor

Simunek J.

autor

Koppova I.

autor

Skalova T.

autor

Stepankova A.

autor

Hasek J.

autor

Dohnalek J.

Bibliografia

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Uwagi

PL

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