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Production and purification of an alkaline lipase from Bacillus sp. for enantioselective resolution of (±)-Ketoprofen butyl ester

Author:
Saraswat, Rashmi, Bhushan, Indu, Gupta, Pankaj, Kumar, Vivek, Verma, Vijeshwar
Source:
3 Biotech 2018 v.8 no.12 pp. 491
ISSN:
2190-572X
Subject:
Bacillus subtilis, Pseudozyma antarctica, agitation, biomass, calcium chloride, carboxylic ester hydrolases, enantioselectivity, ketoprofen, metal ions, molecular weight, olive oil, pH, peptones, pharmaceutical industry, response surface methodology, solvents, surfactants, temperature
Abstract:
The present study was conducted to purify lipase from indigenous Bacillus subtilis strain Kakrayal_1 (BSK-L) for enantioselective resolution of racemic-ketoprofen. The production of lipase (BSK-L) was optimized using Plackett–Burman and central composite design of response surface methodology (RSM). The optimized media containing olive oil (3.5%), MnSO₄ (8 mM), CaCl₂ (5 mM), peptone (20 g/l), pH (8), agitation (180 rpm) and temperature (37 °C) resulted in maximum lipase production of 7500 U/g of cell biomass. The lipase was purified using sequential method to an overall purification fold of 13% with 20% recovery, 882 U/mg specific activity and a molecular weight of 45 kDa. Optimal pH and temperature of purified lipase were found to be 8 and 37 °C, respectively. Furthermore, BSK-L displayed good stability with various organic solvents, surfactants and metal ions. Kₘ and Vₘₐₓ values of lipase were observed to be 2.2 mM and 6.67 mmoles of product formed/min/mg, respectively. The racemic ketoprofen butyl ester was hydrolyzed using lipase with 49% conversion efficiency and 69% enantiomeric excess (ee) which was superior to the commercially procured lipase (Candida antarctica lipase). Thus, this enzyme could be considered as a promising candidate for the pharmaceutical industry.
Agid:
6216149