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Excretome of the chitinolytic bacterium Clostridium paraputrificum J4
- 1.0387248 - ÚŽFG 2013 RIV CZ eng J - Journal Article
Šimůnek, Jiří - Koppová, Ingrid - Tiščenko, Galina - Dohnálek, Jan - Dušková, Jarmila
Excretome of the chitinolytic bacterium Clostridium paraputrificum J4.
Folia Microbiologica. Roč. 57, č. 4 (2012), s. 335-339. ISSN 0015-5632. E-ISSN 1874-9356
R&D Projects: GA ČR GA310/09/1407
Institutional research plan: CEZ:AV0Z50450515; CEZ:AV0Z40500505
Keywords : chitinase * purification * enzymes
Subject RIV: EE - Microbiology, Virology
Impact factor: 0.791, year: 2012
A strictly anaerobic mesophilic chitinolytic bacterial strain identified as Clostridium paraputrificum J4 was isolated from human feces. In response to various types of growth substrates, the bacterium produced an array of chitinolytic enzymes representing significant components of the J4 strain secretome. The excreted active proteins were characterized by estimating the enzymatic activities of endochitinase, exochitinase, and N-acetylglucosaminidase induced by cultivation in medium M-10 with colloidal chitin. The enzyme activities produced by J4 strain cultivated in medium M-10 with glucose were significantly lower. The spectrum of extracellularly excreted proteins was separated by SDS-PAGE. The chitinase variability was confirmed on zymograms of renatured SDS-PAGE. The enzymes were visualized under ultraviolet light by using 4-methylumbelliferyl derivatives of N-acetyl-beta-d-glucosaminide, N,NA '-diacetyl-beta-d-chitobiose, or N,NA ',NEe-triacetyl-beta-d-chitotriose for beta-N-acetylglucosaminidase, chitobiosidase, or endochitinase activities, respectively. Protein components of the secretome were separated by 2D-PAGE analysis. The distinct protein bands were excised, isolated, and subsequently characterized by using MALDI-TOF/TOF tandem mass spectrometry. The final identification was performed according to sequence homology by database searching.
Permanent Link: http://hdl.handle.net/11104/0216465
Number of the records: 1