Extraction and Optimization of Protease Activity by Bacillus Subtitles from Different Sources and Finding out the best Source for Optimized Production in future

Screening and isolation of protease producing strain of bacteria i.e. Bacillus subtilis were carried out by extraction from three different sources i.e. soil, sea water, and degraded abattoir waste from of different locations. This paper gives a study of optimized production of protease from the strains of Bacillus subtilis which shows different criteria of production due to varying sources. Firstly, the protease producing bacterial strain was isolated from sea water and identified by 16S rRNA sequencing. Optimization revealed that the most suitable Nitrogen source was beef extract and carbon source was glucose. 7% NaCl conc. showed highest yield. Most suitable pH was 9 and temperature at 40ºC (1). Secondly, in the study of bacterial strain from degraded abattoir waste, three isolates were found namely yellow-, white- and orange-colored bacteria. Amongst them, white colored colony was found to be more suitable for protease production. The morphological, cultural, biochemical and 16S rRNA identified the bacterial strain. Physical and chemical parameters were optimized for maximum protease production and optimum temperature and pH was found to be 40ºC at pH 14. Glucose as a carbon source and yeast extract as a nitrogen source further stimulated the production process giving maximum protease activity (4). Lastly, Screening and isolation of protease producing strains of bacteria were carried out from four different soil samples collected from various places in Bangalore. The isolates were positive on skim milk agar (1%) and thus are selected as protease producing strain. The organisms were tested for various biochemical tests, which lead to their identification. Optimal growth temperature and pH were found at 37ºC and 8.0 respectively. It was also optimized for carbon test and nitrogen test with optimal growth in dextrose and peptone respectively. Enzyme production was carried in 1 litre of optimized media in the fermenter at 37ºC for 48 hours at pH 8.0. Harvested protease product was purified by salt precipitation method. Finally, the enzyme protease was purified by column chromatography. The protein was characterized using SDS-PAGE. (3) Hence, these results showed that Bacillus subtilis under study is a good producer of protease which can be beneficial for industries in future needs for the protease production.