Abstract
In current study, lipase from a thermotolerant Bacillus subtilis TTP-06 was purified in a stepwise manner by using ammonium sulfate precipitation and column chromatography. Thenceforth, it was subjected to sodium dodecyl sulfate- and native-polyacrylamide gel electrophoresis to check the homogeneity of the purified enzyme. The ideal substrate concentration, pH, temperature, reaction duration and lipase specificity were identified. With a yield of 11.02%, purified lipase displayed activity of 8.51 U/mg. Thenceforward, the homogeneously purified enzyme was considered to be a homo-dimer of 30 kDa subunits. Enzyme had Km and Vmax value of 9.498 mM and 19.92 mol mg−1 min−1, respectively. Additionally, the matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) method was used to investigate the purified lipase and estimate its 3-D structure, which revealed a catalytic triad of serine, aspartate and histidine.
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Acknowledgements
Council of Scientific and Industrial Research (CSIR), Pusa, New Delhi, INDIA, is thankfully acknowledged for providing financial assistance in the form of SRF (Award no.: 09/237(170)/2018-EMR-I). Authors are highly thankful to Department of Biotechnology, Ministry of Science and Technology, Govt. of India, for providing financial support and all necessary facilities to Department of Biotechnology, Himachal Pradesh University, Shimla, India.
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Kaur, M., Kumar, R., Katoch, P. et al. Purification and characterization of extracellular lipase from a thermotolerant strain: Bacillus subtilis TTP-06. 3 Biotech 13, 343 (2023). https://doi.org/10.1007/s13205-023-03717-6
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DOI: https://doi.org/10.1007/s13205-023-03717-6