Applications of A novel alkaline metalloprotease purified by Pseudomonas aeruginosa MH298778
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Abstract
Proteases, an important class of industrially important enzymes, are widely used in detergents, leather processing, silk processing, medicine, food, animal feed, environmental protection and chemical processing. In this study an alkaline protease was isolated from the sediment sample collected from specific site of Lucknow, India and identified as Pseudomonas aeruginosa MH298778 on the basis of 16S rDNA gene sequencing and biochemical properties. The isolated protease was purified to homogeneity using ammonium sulphate precipitation, dialysis and ion exchange chromatography with a fold purification of 2.28 and a recovery of 35.91%. The enzyme has a relative molecular weight of 30 kDa, pH and temperature optima for this protease were 9.0 and 40ᵒC. The activity was stable between a pH ranged of 8.0-10.0. Metal ions such as Ca2+ and Mg2+ stimulated the protease activity up to 115 and 109% respectively. Selected metals stimulated the protease activity where as EDTA completely inhibited indicating it as 'metalloprotease'. The protease was found to be stable in some commonly used commercial detergents tested (Wheel, Rin, Ariel and Tide). It retained 70% residual activity after 3hrs of incubation with Wheel, 71% with Rin, 73% with Ariel and 52% with Tide. The maximum stability was observed with Ariel. The supplementation of the enzyme preparation in detergents could significantly improve the cleansing performance towards the blood stains and suggest its possible use as a laundry additive. The protease was also found efficient of degrading 60% of poultry feathers in 48hrs showing its keratin degrading potential therefore can also be used as an additive in poultry industry in waste treatment.
Key words: Alkaline protease, Purification, Characterization, Detergent compatibility, washing performance, feather degradation, keratin, Pseudomonas aeruginosa MH298778
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