Production and purification of a solvent resistant esterase from Bacillus licheniformis S-86

TORRES SEBASTIAN,; BAIGORI, MARIO D.; ASHOK PANDEY; CASTRO, GUILLERMO R

APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY

Humana Press

Lugar: New York; Año: 2008 vol. 151 p. 221 - 232

0273-2289

New thermophilic and organic-solvent-tolerant Bacillus licheniformis S-86 strain is able to produce two active and solvent-stable esterases. Production of type I and II esterases was substantially enhanced when oils and surfactants were supplied as carbon sources. Grape oil (0.1% v/v) and Tween 20 to 60 (0.1% v/v) had enhanced enzyme production between 1.6 to 2.2-folds. Type II esterase was purified to homogeneity in five-step procedure. This esterase was purified 76.7-folds with a specific activity of 135 U mg-1. Molecular mass of the enzyme was estimated to be 38.4 kDa by SDS-PAGE. Type II esterase was active mostly on esters with short acyl chains, which allowed to classify the enzyme as a carboxylesterase with a Km of 80.2 mmol l-1 and a Vmax of 256.4 μmol min-1 mg-1 for p-nitrophenyl acetate. Also, B. licheniformis S-86 type II esterase displayed activity in presence of water-miscible organic solvents at 50% concentration and stability after one-hour incubation.