Production, optimization and partial purification of protease from terrestrial bacterium Exiguobacterium profundam sp. MM1

A new potent protease producing terrestrial bacterium was isolated and identified as Exiguobacterium profundam sp. MMI by 16 S rRNA gene sequencing. Protease production was enhanced by optimizing the culture conditions. The nutritional factors such as carbon and nitrogen sources, substrate sources and also physical parameters like incubation time, pH, and temperature were optimized for the maximum yield of protease. Studies on the effect of different carbon and nitrogen sources revealed that glucose and peptone enhances the enzyme production. The optimum temperature and pH for enzyme production were at 35 °C and pH 9 respectively. The protease enzyme from Exiguobacterium profundam sp. MMI MG951843.1 was partially purified by ammonium sulphate precipitation and was electrophoresed on 10% (w/v) SDS-PAGE using standard protein markers. The electrophoretic profile of the partially purified enzyme extract shows seven bands with the value of PM obtained from the curve calibration markers to known weight were 48.3KDa. 37.2 kDa, 28.2 kDa, 26.5 kDa, 24.5 kDa,20.4 kDa,16.5 kDa. The band with 48.3KDa shows presence of protease in the sample and further the caseinolytic activity of the partially purified protease enzyme was determined by zymogram.