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Purification and characterization of a cold active alkaline protease from Stenotrophomonas sp., isolated from Kashmir, India

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Title Purification and characterization of a cold active alkaline protease from Stenotrophomonas sp., isolated from Kashmir, India
 
Creator Saba, Iram
Qazi, Parvaiz H.
Rather, Shabir A.
Dar, Refaz A.
Qadri, Qurrat A.
Ahmad, Nasier
Johri, Sarojini
Taneja, Subash C.
Shawl, Sami
 
Subject Biological Sciences
 
Description A Psychrotolerant alkaline protease producing bacterium IIIM-ST045 was isolated from a soil sample collected from the Thajiwas glacier of Kashmir, India and identified as Stenotrophomonas sp. on the basis of its biochemical properties and 16S ribosomal gene sequencing. The strain could grow well within a temperature range of 4–37�C however, showed optimum growth at 15�C. The strain was found to over-produce proteases when it was grown in media containing lactose as carbon source (157.50 U mg-1). The maximum specific enzyme activity (398 U mg-1) was obtained using soya oil as nitrogen source, however, the inorganic nitrogen sources urea,ammonium chloride and ammonium sulphate showed the lowest production of 38.9, 62.2 and 57.9 U mg-1. The enzyme was purified to 18.45 folds and the molecular weight of the partially purified protease was estimated to be *55 kDa by SDS-PAGE analysis. The protease activity increased as the increase in enzyme concentration while as the optimum enzyme activity was found when casein (1% w/v) was used as substrate. The enzyme was highly active
over a wide range of pH from 6.5 to 12.0 showing optimum
activity at pH 10.0. The optimum temperature for the enzyme was 15�C. Proteolytic activity reduced gradually with higher temperatures with a decrease of 56% at 40�C.
The purified enzyme was checked for the removal of protein
containing tea stains using a silk cloth within a temperature range of 10–60�C. The best washing efficiency
results obtained at low temperatures indicate that the
enzyme may be used for cold washing purposes of delicate
fabrics that otherwise are vulnerable to high temperatures.
 
Date 2011
 
Type Article
PeerReviewed
 
Format application/pdf
 
Identifier http://iiim.csircentral.net/321/1/10.1007s11274-011-0905-1.pdf
Saba, Iram and Qazi, Parvaiz H. and Rather, Shabir A. and Dar, Refaz A. and Qadri, Qurrat A. and Ahmad, Nasier and Johri, Sarojini and Taneja, Subash C. and Shawl, Sami (2011) Purification and characterization of a cold active alkaline protease from Stenotrophomonas sp., isolated from Kashmir, India. World Journal of Microbiology and Biotechnology. ISSN 0959-3993
 
Relation http://dx.doi.org/10.1007/s11274-011-0905-1
http://iiim.csircentral.net/321/