Purification of Protease from Pseudomonas thermaerum XXXX Isolated from Poultry Waste Site.

@entity8862 @entity10694 @entity8798 @entity1025 @entity8863 @entity4859 @entity3281 @entity1046 @entity1042 @entity1324 @entity653 @entity23593 @entity3733 @entity7935 @entity141 @entity5723
An extracellular protease was purified from Pseudomonas thermaerum @entity23593 a new strain identified by morphological, biochemical and 16S rDNA sequencing. It was isolated from soil of Poultry waste site at Ghazipur near Ghaziabad, Delhi. The strain produces extra cellular protease in the culture media that was maintained at 37  C, 140 rpm. The media was harvested for protease after 48 hrs of incubation at 37  C in basal media supplemented with 1% casein. We report 6.08 fold purification of enzyme following @entity4859 precipitation and DEAE-cellulose chromatography. The molecular weight of the enzyme was estimated to be approximately 43,000 daltons as shown by casein zymography studies. The optimum pH for the proteolytic activity was pH 8.0 and enzyme remained stable between pH 5 -11 at 60  C. Interestingly @entity3281 (5mM) activated enzyme activity by 5 fold, while @entity8863 , @entity1025 and @entity141 moderately activated enzyme activity, where as @entity8862 , @entity8798 and @entity10694 inhibited enzyme activity. The protease produced was stable in presence of 50 % (v/v) @entity1046 and @entity1324 . @entity5723 , @entity1042 and @entity7935 increased protease activity by 2.7, 1.3 and 1.1 fold respectively but was inhibited in presence of @entity653 and @entity3733 . This organic solvent-stable protease could be used as a biocatalyst for enzymatic peptide synthesis.
