Production and Immobilization of Alpha Amylase by Using Bacillus subtilis

Abstract

The α-amylase producing organisms were isolated from soil sample, such as Bacillus sp (2 isolates), Bacillus cereus (1 isolates), Bacillus subtilis (2 isolates) by basal medium and identified by standard biochemical test. The high yielding strain was identified as Bacillus subtilis and used for amylase production. Different  parameters  like temperature (10°C, room temperature (25°-30°C), 37°C & 55°C different pH (6.0, 6.5, 7.0, 7.5, 8.0) and incubation periods (1-5 days) were used for the α-amylase production. The isolate was inoculated to the production medium and incubated, then it was centrifuged  and the supernatant containing crude enzyme extract is used for protein estimation and activity. It was found that α-amylase production and activity was high at 37°Cat pH 7.0 in 48 hours. The organism was grown in the optimum conditions mentioned above and the enzyme α-amylase was immobilized by alginate gel entrapment method. From the organism the plasmid and the chromosomal DNA was isolated and detected.