ISSN 2756-3391
African Journal of Parasitology Research ISSN 2756-3391 Vol. 12 (9), pp. 001-006, September, 2024. Available online at www.internationalscholarsjournals.org © International Scholars Journals
Full Length Research paper
Extraction, Purification, and Scalable Production of Protease Enzyme from bacillus subtilis
Gitishree Das1* and M.P. Prasad2
1Biotechnology Laboratory, Crop Improvement Division, Central Rice Research Institute, Cuttack, Orissa, India
2Sangenomics Research Laboratory, Bangalore
Accepted 26 April, 2024
Abstract
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 as Bacillus subtilis producing protease enzyme. These Bacillus subtilis could group upto 40ºC and pH range 6-9 with optimal growth temperature and pH 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. This result showed that Bacillus subtilis under study is a good producer of extra cellular protease, which can be beneficial for industries.
Keywords: Bacillus subtilis, protease, fermenter, chromatography.