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Extracellular solvent stable cold-active lipase from psychrotrophic Bacillus sphaericus MTCC 7526: partial purification and characterization

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Abstract

Psychrotropic Bacillus sphaericus producing solvent stable cold-active lipase upon growth at low temperature was isolated from Gangotri glacier. Optimal parameters for lipase production were investigated and the strain was able to produce lipase even at 15 °C. An incubation period of 48 h and pH 8 was found to be conducive for cold-active lipase production. The addition of trybutyrin as substrate and lactose as additional carbon source increased lipase production. The enzyme was purified up to 17.74-fold by ammonium sulphate precipitation followed by DEAE cellulose column chromatography. The optimum temperature and pH for lipase activity were found to be 15 °C and 8.0, respectively. The lipase was found to be stable in the temperature range 20–30 °C and the pH range 6.0–9.0. The protein retained more than 83 % of its initial activity after exposure to organic solvents. The lipase exhibited significant stability in presence of acetone and DMSO retaining >90 % activity. The enzyme activity was inhibited by 10 mM CuSO4 and EDTA but showed no loss in activity after incubation with other metals or inhibitors examined in this study.

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Acknowledgement

The authors are grateful to Department of Biotechnology Govt. of India for the grant (BT/PR2639/PID/24/112/2001) received to carry out this study.

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  1. Babu Joseph

    Present address: College of Applied Medical Sciences, Shaqra University, P.B. No. 1383, Shaqra, Kingdom of Saudi Arabia

Authors and Affiliations

  1. Sam Higginbottom Institute of Agriculture, Technology and Sciences, Allahabad, 211 007, Uttar Pradesh, India

    Babu Joseph

  2. Department of Biological Sciences, School of Basic Sciences, Sam Higginbottom Institute of Agriculture, Technology and Sciences, Allahabad, 211 007, Uttar Pradesh, India

    Pramod W. Ramteke

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  1. Babu Joseph
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Correspondence to Babu Joseph.

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Joseph, B., Ramteke, P.W. Extracellular solvent stable cold-active lipase from psychrotrophic Bacillus sphaericus MTCC 7526: partial purification and characterization. Ann Microbiol 63, 363–370 (2013). https://doi.org/10.1007/s13213-012-0483-y

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  • DOI: https://doi.org/10.1007/s13213-012-0483-y

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