OPTIMIZATION OF PRODUCTION PARAMETERS FOR ALKALINE PROTEASE PRODUCTION FROM BACILLUS SPP.

Kanupriya Miglani, Jaswinder Kaur and Rajesh kumar* * Corresponding author: email: dahiya76@gmail.com Department of Biotechnology, University Institute of Engineering & Technology, Kurukshetra University Kurukshetra, Haryana ABSTRACT

Proteases are the enzymes that catalyse hydrolytic reactions in which protein molecules are degraded to peptides and amino-acids. Microbial proteases are extracellular in nature and are directly secreted into the fermentation broth by the producer, thus simplifying downstream processing of the enzyme as compared to the proteases obtained from plants and animals. Alkaline proteases have attracted attention due to their multitude industrial applications such as fertilizers, detergents, leather, pharmaceutical and biotechnological applications such as peptide synthesis, silver recovery from used X-ray or photographic films and proteinaceous fodder from waste feathers or keratin containing materials. In the present work alkaline protease producing bacteria Bacillus K3, a close relative of Bacillus ayrabhattai was isolated from the soil samples collected from poultry waste dumping site, areas near butcher shop and milk vendors in Kurukshetra and Ludhiana and the parameters for the production of enzyme were optimized. Out of all carbon sources studied a concentration of 10.0 g/l of lactose gave the maximum protease titre (625.73U/ml). Evaluation of nitrogen sources illustrated that yeast extract and casein when used in combination at a concentration of 20.0 g/l (10.0 g/l each) gave the maximum production with the protease titre of 779.29 U/ml. Out of various salts studied CaCl2 when used at the concentration of 4.0 g/l gave the maximum protease titre (950.36 U/ml). Production was increased when the inoculum size was varied from 1% to 5% (v/v) and it decreased with further increase in the inoculum size. The pH and temperature were optimized to 8 and 37 C respectively for the maximum production. Finally the production carried out under fully optimized conditions resulted in 6.66 fold increase in alkaline protease production.