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Cloning, expression and characterization of a novel esterase from Bacillus pumilus

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Abstract

The gene encoding esterase (CE1) from Bacillus pumilus ARA with a calculated molecular weight of 28.4 kDa was cloned, sequenced and efficiently expressed in Escherichia coli. The open reading frame of 747 nucleotides encoded a protein, which was classified as a carboxylesterase with an identity of 87 % to esterase from Bacillus subtilis 168. Recombinant CE1 was purified in a single step to electrophoretic homogeneity by IMAC (Ni2+). The enzyme displayed maximum activity toward p-nitrophenyl (pNP) acetate at 37–40 °C and pH 6.5–7.0. It was stable in the pH range from 6.5 to 8.0, and at temperature from 25 to 40 °C. Among four p-nitrophenyl esters tested, the best substrate was pNP acetate with K m and k cat values of 0.33 mM and 4.07 s−1, respectively. Amounts of 2 mM Ca2+ and Co2+ significantly increased the esterase activity to 190 and 121 %, respectively. These results suggest that CE1 has very attractive applications of increasing feed digestibility in animal nutrition in this moderate temperature range.

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Acknowledgments

We thank Prof. Weilan Shao and Dr. Jianjun Pei for providing Bacillus pumilus ARA. This work was supported by grants from NSF of Jiangsu Province of China (BK2010545).

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Authors and Affiliations

  1. Department of Food Science and Nutrition, GinLing College, Nanjing Normal University, Nanjing, People’s Republic of China, 210097

    Yemin Xue

  2. Department of Microbiology, College of Life Science, Nanjing Normal University, Nanjing, People’s Republic of China, 210046

    Jingjing Zhang, Xixi Wu, Huihui Sun & Chunying Xu

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  1. Yemin Xue
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  2. Jingjing Zhang
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Correspondence to Yemin Xue.

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Xue, Y., Zhang, J., Wu, X. et al. Cloning, expression and characterization of a novel esterase from Bacillus pumilus . Ann Microbiol 63, 879–885 (2013). https://doi.org/10.1007/s13213-012-0539-z

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  • DOI: https://doi.org/10.1007/s13213-012-0539-z

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