Asian Journal of Microbiology, Biotechnology & Environmental Sciences Paper

Vol 17, Issue 1, 2015; Page No.(79-85)

PURIFICATION AND CHARACTERIZATION OF AMIDASE FROM BACILLUS SP. APB-6.

SUDHA BATTA, POONAM KUMARI AND DUNI CHAND

Abstract

Amidases (EC 3.5.1.4) hydrolyzes amides with the production of the corresponding carboxylic acids and ammonia and are the most widely used amide-hydrolyzing enzymes in industry. It holds a great potential for biotechnological applications in organic acids synthesis, bioremediation, waste water treatment, synthesis of pharmaceutical chemical like anti-cancerous and anti-HIV Hydroxamic acids etc. Here, we report the purification of amidase from Bacillus sp. APB-6 and after purification the enzyme was characterized for optimum activity. The enzyme was purified by about 21.7 fold with specific activity 19.5 U/mg protein. A purified amidase enzyme of 44kDa was obtained after initially precipitated with 60% Acetone. The acetone precipitated enzyme was then loaded on to the anion exchange column (EAECellulose) and then the protein was analysed by SDS-PAGE. The optimum temperature and pH of the enzyme was 55pC and 7.5 respectively. The Km and Vmax values for the purified amidase of APB-06 were 200mM and the Vmax was 100 μmoles/mg/min The enzyme was highly specific to substrate acetamide and less activity was detected in the presence of other amides, such as butyramide, cyanamide, nicotinamide and benzamide.

Enter your contact information below to receive full paper.
Your Name :
Email:
Phone:
City:
Cost of Full Paper: Rs.75 for Indian Nationals or $20 (USD) for international subscribers.
By clicking on Request Paper you Agree to pay the above mentioned cost per paper.