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Solvent-Tolerant Acyltransferase from Bacillus sp. APB-6: Purification and Characterization

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Abstract

Amidase from Bacillus sp. APB-6 with very good acyltransferase activity was purified to homogeneity with a purification fold of 3.68 and 53.20% enzyme yield. The purified protein's subunit molecular mass was determined approximately 42 kDa. Hyperactivity of the enzyme was observed at pH 7.5 (150 mM, potassium-phosphate buffer) and 50 °C of incubation. An enhancement in activity up to 42% was recorded with ethylenediaminetetraacetic acid and dithiothreitol. The kinetic parameter Km values for substrates: acetamide and hydroxylamine-hydrochloride were 73.0 and 153 mM, respectively. Further, the Vmax for acyltransferase activity was 1667 U/mg of protein and the Ki for acetamide was calculated as 37.0 mM. The enzyme showed tolerance to various organic solvents (10%, v/v) and worked well in the biphasic reaction medium. The acyltransferase activity in presence of solvents i.e. biphasic medium may prove highly favorable for the transformation of hydrophobic amides, which otherwise is not possible in simple aqueous phase.

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Acknowledgements

Department of Biotechnology, Himachal Pradesh University, Shimla is duly acknowledged for extending their research laboratory facility.

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Authors and Affiliations

  1. Department of Reproductive Biology, All India Institute of Medical Sciences, New Delhi, 110029, India

    Deepak Pandey

  2. Department of Biotechnology, Himachal Pradesh University, Shimla, HP, 171005, India

    Sanjay K. S. Patel, Rajendra Singh, Vikram Thakur & Duni Chand

  3. Faculty of Applied Sciences and Biotechnology, Shoolini University, Solan, HP, 173229, India

    Pradeep Kumar

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  1. Deepak Pandey
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Pandey, D., Patel, S.K.S., Singh, R. et al. Solvent-Tolerant Acyltransferase from Bacillus sp. APB-6: Purification and Characterization. Indian J Microbiol 59, 500–507 (2019). https://doi.org/10.1007/s12088-019-00836-8

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