Production and Partial Characterization of Extracellular Protease from Bacillus sp. GJP2 Isolated from a Hot Spring

Abstract

In the present investigation extracellular protease production is reported from a moderately thermotolerant bacterial strain GJP2, isolated from a hot spring. The enzyme production conditions were optimized by determining maltose as the best carbon source and a combination of yeast extract and peptone as the most suitable nitrogen sources at pH 8.5 and temperature of 50 °C. A total of 31.7 fold enzyme purification was achieved by precipitating the culture supernatant with 70 % (NH4)2SO4 followed by Sephadex G-100 and ion exchange chromatography. Electrophoresis analysis of the purified enzyme revealed its molecular weight approximately as 64 kDa. The purified enzyme showed maximum activity at pH and temperature of 8.5 and 50 °C, respectively. Based on the partial 16S rRNA sequencing and biochemical properties the bacterium was identified as a Bacillus sp.