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Identification and characterization of a novel cold-adapted esterase from a metagenomic library of mountain soil

  • Biocatalysis
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

A novel lipolytic enzyme was isolated from a metagenomic library after demonstration of lipolytic activity on an LB agar plate containing 1% (w/v) tributyrin. A novel esterase gene (estIM1), encoding a lipolytic enzyme (EstIM1), was cloned using a shotgun method from a pFosEstIM1 clone of the metagenomic library, and the enzyme was characterized. The estIM1 gene had an open reading frame (ORF) of 936 base pairs and encoded a protein of 311 amino acids with a molecular mass 34 kDa and a pI value of 4.32. The deduced amino acid sequence was 62% identical to that of an esterase from an uncultured bacterium (ABQ11271). The amino acid sequence indicated that EstIM1 was a member of the family IV of lipolytic enzymes, all of which contain a GDSAG motif shared with similar enzymes of lactic acid microorganisms. EstIM1 was active over a temperature range of 1–50°C, at alkaline pH. The activation energy for hydrolysis of p-nitrophenyl propionate was 1.04 kcal/mol, within a temperature range of 1–40°C. The activity of EstIM1 was about 60% of maximal even at 1°C, suggesting that EstIM1 is efficiently cold-adapted. Further characterization of this cold-adapted enzyme indicated that the esterase may be very valuable in industrial applications.

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Acknowledgments

This work was supported by an HTS-based Integrated Technology Development grant (no. 2008-04171) from the Ministry of Education, Science and Technology through the National Research Foundation of Korea, and a basic research grant from KRIBB.

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Authors and Affiliations

  1. Microbe-based Fusion Technology Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), 1404 Sinjeong-dong, Jeongeup, Jeonbuk, 580-185, Republic of Korea

    Kyong-Cheol Ko, Soon-Ok Rim, Yunjon Han, Bong Seok Shin, Jong Hyun Choi & Jae Jun Song

  2. Department of Biological Sciences, College of Natural Sciences, Chonnam National University, 300 Yong-Bong Dong, Buk-Gu, Gwangju, 500-757, Republic of Korea

    Bong Seok Shin & Geun-Joong Kim

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  1. Kyong-Cheol Ko
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  2. Soon-Ok Rim
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Correspondence to Jong Hyun Choi or Jae Jun Song.

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Ko, KC., Rim, SO., Han, Y. et al. Identification and characterization of a novel cold-adapted esterase from a metagenomic library of mountain soil. J Ind Microbiol Biotechnol 39, 681–689 (2012). https://doi.org/10.1007/s10295-011-1080-y

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  • DOI: https://doi.org/10.1007/s10295-011-1080-y

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