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Characterization and a point mutational approach of a psychrophilic lipase from an arctic bacterium, Bacillus pumilus

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Abstract

A bacterium with lipolytic activity was isolated from the Chukchi Sea within the Arctic Ocean. The lipase BpL5 from the isolate, Bacillus pumilus ArcL5, belongs to subfamily 4 of lipase family I. The optimum pH and temperature of the recombinant enzyme BpL5, as expressed in Escherichia coli, were 9.0 and 20 °C, respectively. The enzyme retained 85 % of its activity at 5 °C. There was a significant difference between temperatures for maximal activity (20 °C) and for protein denaturation (approx. 45 °C). The enzyme preferred middle-chain (C8) p-nitrophenyl substrates. Two mutants, S139A and S139Y, were rationally designed based on the 3D-structure model, and their activities were compared with that of the wild type. The both mutants showed significantly improved activity against tricaprylin.

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Acknowledgments

Experiment of Circular Dichroism spectroscopy was carried out by Mr. Hackwon Do. This research was supported by a Grant from the Ministry of Oceans and Fisheries (PM12030).

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Authors and Affiliations

  1. Division of Life Sciences, Korea Polar Research Institute (KOPRI), 26, Songdomirae-ro, Yeonsu-gu, Incheon, 406-840, Republic of Korea

    Ah Ram Wi, Sunghui Kim, Ha Ju Park, Dockyu Kim, Se Jong Han, Joung Han Yim & Han-Woo Kim

  2. Department of Biotechnology & Bioengineering, Dong-Eui University, 995 Eomgwangno, Busanjin-gu, Busan, 614-714, Republic of Korea

    Sung-Jong Jeon

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  1. Ah Ram Wi
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  3. Sunghui Kim
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  4. Ha Ju Park
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  5. Dockyu Kim
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  6. Se Jong Han
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  8. Han-Woo Kim
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Correspondence to Han-Woo Kim.

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Wi, A.R., Jeon, SJ., Kim, S. et al. Characterization and a point mutational approach of a psychrophilic lipase from an arctic bacterium, Bacillus pumilus . Biotechnol Lett 36, 1295–1302 (2014). https://doi.org/10.1007/s10529-014-1475-8

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  • DOI: https://doi.org/10.1007/s10529-014-1475-8

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