Production and purification of a solvent-resistant esterase from Bacillus licheniformis S-86.
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- and 2.2-folds. Type II esterase was purified to homogeneity in a five-step procedure. This esterase was purified 76.7-fold with a specific activity of 135 U mg(-1). Molecular mass of the enzyme was estimated to be 38.4 kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Type II esterase was active mostly on esters with short acyl chains, which allowed to classify the enzyme as a carboxylesterase with a K (m) of 80.2 mmol l(-1) and a V (max) of 256.4 micromol 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 1-h incubation.
Planta Piloto de Procesos Industriales Microbiológicos PROIMI, Tucumán, Argentina.
Sebastián
S
Torres
Mario D
MD
Baigorí
Ashok
A
Pandey
Guillermo R
GR
Castro
D001407:Bacillus / Q000187:drug effects / Q000201:enzymology*; D002265:Carboxylic Ester Hydrolases / Q000096:biosynthesis* / Q000187:drug effects / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D007700:Kinetics; D010938:Plant Oils / Q000494:pharmacology; D011136:Polysorbates / Q000494:pharmacology; D012997:Solvents / Q000494:pharmacology; D013379:Substrate Specificity; D013501:Surface-Active Agents / Q000494:pharmacology; D027843:Vitis
D010938:Plant Oils; D011136:Polysorbates; D012997:Solvents; D013501:Surface-Active Agents; D002265:Carboxylic Ester Hydrolases