Iranian Journal of Biotechnology, ( ISI ), Volume (10), No (2), Year (2012-3) , Pages (120-128)

Title : ( Purification and characterization of a thermostable neutrophilic metalloprotease from Pseudomonas sp. DR89 )

Authors: Ahmad Asoodeh , hossein mohammadian musa abadi ,

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A novel neutrophilic metalloprotease was isolated from Pseudomonas sp. DR89 isolate which was identified in a mineral spring in Iran. The enzyme was purified from the isolate to 21-folds in a three-step procedure involving ammonium sulfate precipitation, Q-Sepharose ionic exchange and Sephadex G-100 gel filtration chromatography. Resuts showed that the enzyme was active at high temperatures and in a wide-range pH of 5-11 with the optimum of 8.0. The zymogram and sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis revealed the presence of one protease with a molecular weight of 74 kDa. The enzyme activity was decreased by Zn2+, Mn2+, H2O2 and cetyl trimethylammonium bromide (CTAB), whereas its activity was increased by Ca2+, Mg2+, Cu2+ and dimethyl sulfoxide (DMSO). Na+, phenylmethyl sulfonylfluoride (PMSF), β-mercaptoethanol, sodium dodecyl sulfate (SDS), and Triton X-100 did not show a considerable effect on its activity. Casein was a better substrate than bovin serum albumin (BSA) and gelatin for this enzyme. The kinetic parameters (Km and Vmax) of the purified protease towards caseinolytic activity were also determined. These properties of the enzyme make it suitable for use in food industries.

Keywords

Neutrophilic protease; phylogenetic tree; thermostability; purification;
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@article{paperid:1027270,
author = {Asoodeh, Ahmad and Mohammadian Musa Abadi, Hossein},
title = {Purification and characterization of a thermostable neutrophilic metalloprotease from Pseudomonas sp. DR89},
journal = {Iranian Journal of Biotechnology},
year = {2012},
volume = {10},
number = {2},
month = {March},
issn = {1728-3043},
pages = {120--128},
numpages = {8},
keywords = {Neutrophilic protease; phylogenetic tree; thermostability; purification; characterization},
}

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%0 Journal Article
%T Purification and characterization of a thermostable neutrophilic metalloprotease from Pseudomonas sp. DR89
%A Asoodeh, Ahmad
%A Mohammadian Musa Abadi, Hossein
%J Iranian Journal of Biotechnology
%@ 1728-3043
%D 2012

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