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


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. K m and V max 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.

SUBMITTER: Saraswat R 

PROVIDER: S-EPMC6242800 | biostudies-literature | 2018 Dec

REPOSITORIES: biostudies-literature

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

Saraswat Rashmi R   Bhushan Indu I   Gupta Pankaj P   Kumar Vivek V   Verma Vijeshwar V  

3 Biotech 20181119 12


The present study was conducted to purify lipase from indigenous <i>Bacillus subtilis</i> 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%), MnSO<sub>4</sub> (8 mM), CaCl<sub>2</sub> (5 mM), peptone (20 g/l), pH (8), agitation (180 rpm) and temperature (37 °C) resulted in maximum lip  ...[more]

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2014-06-17 | GSE38276 | GEO