Monday, July 27, 2009 - 8:50 AM
S18

Isolation of Protease and Keratinase From Microbes Isolated From Ghazipur Poultry Waste Site, Ghaziabad, India

Smriti Gaur, Sarita Gupta, and Neeraj Wadhwa. Biotechnology, Department of Biotechnology, Jaypee Institute of Information Technology University, A-10 sec 62, ,, Noida, India

Feathers are produced in large amount as waste by poultry product processing plants. Structural keratin can be degraded by some proteolytic micro-organisms which degrade keratin after hydrolysis, the feather can be converted to feed stuffs, fertilizers, glues & films. The  soil sample was collected from Ghazipur poultry waste site, Ghaziabad, India were reinoculated in enrichment medias Horikoshi media (I), Feather meal media 1 (II), Feather meal media 2 (III). Culture supernatant were harvested and checked for proteases and keratinases activity using appropriate substrates. The optimal medium for enzyme production is media III. Further synthesis of keratinases and proteases was done in the selection media. We report that keratinase was purified 1.463 fold after 0-80% ammonium sulphate precipitation. Colonies producing clear zone in feather meal agar were selected and identified as GW2, GW3 and GW4 were able to degrade chicken and pigeon feathers whereas only one colony GW1 was showed degradation on skim milk agar plates. This strain could not degrade the chicken and pigeon feathers but was able to produced protease.The strain GW1 produces protease specifically in the culture media that was maintained at 37°C and at 140 rpm for 48 hrs. We report 2.33 fold purification of enzyme following 0-60% ammonium sulphate precipitation. The optimum pH for the proteolytic activity was pH 8.0 and enzyme remained stable between pH 5 -11 at 60°C. Interestingly Mn2+ (5mM)  strongly activated enzyme activity by 5 fold.

 


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