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Purification and characterization of an extracellular haloalkaline protease produced by the moderately halophilic bacterium, Salinivibrio sp. strain AF-2004

Paper ID Volume ID Publish Year Pages File Format Full-Text
18288 42717 2007 7 PDF Available
Title
Purification and characterization of an extracellular haloalkaline protease produced by the moderately halophilic bacterium, Salinivibrio sp. strain AF-2004
Abstract

Among several strains of moderately halophilic bacteria that were isolated from various areas in Iran, strain AF-2004 was selected as the best producer of extracellular protease and was used for further studies. Phenotypic classification and 16S rRNA sequence analysis placed AF-2004 in the genus Salinivibrio. Maximal protease production was detected at the end of exponential growth phase in the medium containing 5% (w/v) NaCl. This protease was purified to homogeneity by a combination of acetone precipitation, Q-Sepharose ion exchange and Sephacryl S-200 gel filtration chromatography. The enzyme was a monomeric protease with a relative molecular mass of 38–43 kDa by SDS-PAGE and gel filtration chromatography. The specific activity of the purified enzyme was determined to be 171.6 μmol of tyrosine/min per mg of protein using casein as a substrate. The enzyme exhibited its optimal activity at 65 °C, pH 8.5, and 0–0.5 M NaCl with a high tolerance to salt concentrations of up to 4 M. The protease was identified as a zinc-metalloprotease, which was strongly inhibited by EDTA and 1,10-phenanthroline, and the N-terminal amino acid sequence determined showed high similarity to the zinc-metalloproteases from Vibrio species.

Keywords
Moderately halophilic bacteria; Salinivibrio; Haloalkaline protease; Zinc-metalloprotease
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Purification and characterization of an extracellular haloalkaline protease produced by the moderately halophilic bacterium, Salinivibrio sp. strain AF-2004
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Publisher
Database: Elsevier - ScienceDirect
Journal: Enzyme and Microbial Technology - Volume 40, Issue 2, 4 January 2007, Pages 266–272
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us