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2011 | 58 | 2 | 261-263
Article title

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

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EN
Abstracts
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.
Publisher

Year
Volume
58
Issue
2
Pages
261-263
Physical description
Dates
published
2011
received
2011-03-05
revised
2011-05-12
accepted
2011-06-08
(unknown)
2011-06-13
Contributors
  • Institute of Macromolecular Chemistry, Academy of Sciences of the Czech Republic, Praha, Czech Republic
  • Institute of Macromolecular Chemistry, Academy of Sciences of the Czech Republic, Praha, Czech Republic
  • Institute of Macromolecular Compounds, Russian Academy of Sciences, St. Petersburg, Russia
  • Institute of Animal Physiology and Genetics, Academy of Sciences of the Czech Republic, Praha, Czech Republic
  • Institute of Animal Physiology and Genetics, Academy of Sciences of the Czech Republic, Praha, Czech Republic
  • Institute of Macromolecular Chemistry, Academy of Sciences of the Czech Republic, Praha, Czech Republic
  • Institute of Macromolecular Chemistry, Academy of Sciences of the Czech Republic, Praha, Czech Republic
  • Institute of Animal Physiology and Genetics, Academy of Sciences of the Czech Republic, Praha, Czech Republic
author
  • Institute of Macromolecular Chemistry, Academy of Sciences of the Czech Republic, Praha, Czech Republic
References
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Document Type
Publication order reference
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YADDA identifier
bwmeta1.element.bwnjournal-article-abpv58p261kz
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