European Journal of Biomedical and Pharmaceutical Sciences

PRODUCTION AND PURIFICATION OF ?-AMYLASE FROM BACILLUS SPECIES ISOLATED FROM SOIL USING AGRO WASTE RICE HUSK IN SOLID STATE FERMENTATION

M. Shakti Chandra Vadhana, T. Monica and G. Sona*

ABSTRACT

Amylase is an enzyme that catalyzes the breakdown of starch into sugars. The amylases obtained from microorganisms have a broad spectrum of industrial uses as they are more stable than plant and animal amylases. The major advantage of using microorganisms for the production of amylases is the economical bulk production capacity, and also microbes are easy to manipulate to derive enzymes of desired nature. The present study focuses on the isolation of amylase producing Bacillus species from soil sample and production of bacterial amylase. In this study, we have also compared the amylase produced in normal basal medium and in media in which rice husk has been given as a specific substrate with basal medium. The amylase enzyme produced was estimated by DNS method, in which it was observed that when rice husk was used as a substrate it gave 59% higher production of amylase compared to the normal basal medium. The amylase produced in normal basal medium was 7.27 U/ml whereas amylase produced in basal medium with rice husk as specific substrate was 18.1 U/ml. The amylase enzyme was then extracted from the medium by ammonium sulfate precipitation method and purified by dialysis. For further analysis and characterization, the enzyme was subjected to SDS-PAGE analysis in which molecular weight of amylase was determined to be approximately 50 kDa. Hence it was concluded that agro waste rice husk can be used as an effective and economical substrate for the production of microbial amylase.

Keywords: ?-Amylase, Bacillus subtilis, Dinitrosalicylic acid, agro waste, rice-husk.