Molecular identification of newly isolated bacillus strains from poultry farm and optimization of process parameters for enhanced production of extracellular amylase using OFAT method

Abstract

Bacillus sp. has been used as attractive and efficient industrial organisms in the last few decades due to their rapid growth rates and their potential to secrete extracellular enzymes into the medium. Amylase obtained from bacteria has gained immense interest due to its biotechnological potential in various industries. The aim of this experimental context was to isolate novel strains of Bacillus sp. from poultry farm, screening and optimization of different parameters for the enhanced production of extracellular amylase using traditional OFAT method (One Factor At a Time). Five hyperamylase producing Bacillus strains were identified after preliminary screening of poultry feces soil sample. The strains were showing maximum amylase production at pH ranging from 6-9. On the other hand strains were showing more enzyme activity at 35 and 45°C. The agitation speed of 130 and 150 rpm was found to be suitable for maximum enzyme production by Bacillus sp. The amylase activity of each isolate was reduced with increase in incubation time from 24-96 h. Starch and yeast extract were found to be the suitable carbon and nitrogen source in order to enhance the enzyme production. The enzyme obtained from the isolates was stable upto 60°C and pH 9.0 for 4 h of incubation. Percentage GC-content of 16S rRNA sequences showed a significant effect on amylase production at high temperature. The present study clearly indicates that amylase production was 2 fold higher when optimized conditions were used. The crude amylase obtained from the isolates showed stability at high temperature and pH. As to the best of our knowledge this present investigation makes a novel contribution to research on amylase production from new Bacillus strains of poultry farm by demonstrating the isolation, molecular characterization and enhanced amylase activity at optimal conditions using OFAT method.