Purification and characterization of an extreme halothermophilic protease from a halophilic bacterium Chromohalobacter sp. TVSP101
Braz. j. microbiol
;
40(1): 12-19, Jan.-Mar. 2009. ilus, graf, tab
Article
in English
| LILACS
| ID: lil-513109
ABSTRACT
An extreme halophilic bacterium was isolated from solar saltern samples and identified based on biochemical tests and 16S r RNA sequencing as Chromohalobacter sp. strain TVSP101. The halophilic protease was purified using ultrafiltration, ethanol precipitation, hydrophobic interaction column chromatography and gel permeation chromatography to 180 fold with 22% yield. The molecular mass of the protease determined by SDS PAGE was 66 kDa. The purified enzyme was salt dependent for its activity and stability with an optimum of 4.5 M NaCl. The optimum temperature for maximum protease activity was 75ºC. The protease was optimally active at pH 8 and retained more than 80% of its activity in the range of pH 7-10. Sucrose and glycine at 10% (w/v) were the most effective osmolytes, retained 100% activity in the absence of NaCl. The activity was completely inhibited by ZnCl2 (2 mM), 0.1% SDS and PMSF (1mM). The enzyme was not inhibited by 1mM of pepstatin, EDTA and PCMB. The protease was active and retained 100% it activity in 10% (v/v) DMSO, DMF, ethanol and acetone.
Full text:
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Index:
LILACS (Americas)
Main subject:
Peptide Hydrolases
/
Solvents
/
Euryarchaeota
/
Halomonadaceae
/
Enzyme Activation
Language:
English
Journal:
Braz. j. microbiol
Journal subject:
Microbiology
Year:
2009
Type:
Article
Affiliation country:
India
Institution/Affiliation country:
Gulbarga University/IN
/
Pune University/IN
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