ISOLATION, PRODUCTION AND REACTION CONDITIONS OPTIMIZATION OF AMIDASE OF BACILLUS SP. MNB1

Abstract

Amidase (EC 3.5.1.4) is a nitrile metabolizing enzyme, belongs to the family of hydrolases, which acts on carbon-nitrogen bonds other than peptide bonds. It usually occurs in prokaryotic and eukaryotic as an inducible intracellular enzyme, which is involved in nitrogen metabolism. It catalyzes hydrolysis of amide to carboxylic acid and ammonium. Amidases find wide applications in various industries as commodity chemicals (e.g. acrylic acid, nicotinic acid etc.) or pharmaceutically important acids (organic acids and hydroxamic acids). In current reasearch project the isolation, production and reaction conditions optimization of mesophilic amidase were undertaken. An aliphatic selective amide degrading gram positive Bacillus sp. MNB1 strain was isolated from thermal spring soil of Manikaran, Himachal Pradesh, India. Amidase production and reaction conditions were optimized. Amidase of Bacillus sp. MNB1 exhibited maximum biomass and activity in M2 medium at temperature 37ºC, pH 7.0 in the presence of acetamide as inducer. The enzyme showed maximum activity at pH 7.0, temperature 37 ºC and 40-50 mM of acetamide as substrate. The enzyme exhibited activity in 30-50% cut of ammonium sulphate precipitation and eluted fraction 0.1-0.5M exhibited amidase activity (secondary activity of amidase) confirmed its presence in the strain Bacillus sp. MNB1.