Author(s):

Sanjay Kumar Yadav, Poonam Singh, Kashyap Kumar Dubey and Bhanu P Singh

Abstract: Partial purification of crude enzyme was carried out by ammonium sulphate precipitation, dialysis and ion exchange chromatography. Alkaline protease was purified upto 2.19 fold as compared with crude enzyme. The apparent molecular weight of partially purified enzyme was determined as 48 kDa by SDS-PAGE and the same was confirmed from zymography. The maximum alkaline protease production was achieved with 7% inoculums size at pH 10, 36 ºC incubation temperature, with impellor tip velocity of 4714 cm/min, and 48hrs of incubation period. Alkaline protease activity was completely inhibited by PMSF; while EDTA showed no inhibition on protease activity that is suggested that the enzyme is a serine protease. The Km and Vmax for the partially purified alkaline protease were 4.343mg and 1mg/min^-1ml^-1 respectively. Alkaline protease was used for hide deharing of goat and found that complete deharing was achieved with protease unit 200 U/ml and in 6 hrs having Mn²⁺ concentration of 10 mM with 50 ⁰C temperature and 10pH

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