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Characterization of Novel Family IV Esterase and Family I.3 Lipase from an Oil-Polluted Mud Flat Metagenome

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Abstract

Two genes encoding lipolytic enzymes were isolated from a metagenomic library constructed from oil-polluted mud flats. An esterase gene, est3K, encoded a protein of 299 amino acids (ca. 32,364 Da). Est3K was a family IV esterase with typical motifs, HGGG, and HGF. Although est3K showed high identity to many genes with no information on their enzymatic properties, Est3K showed the highest identity (36 %) to SBLip5.1 from forest soil metagenome when compared to the enzymes with reported properties. A lipase gene, lip3K, encoded a protein of 616 amino acids (ca. 64,408 Da). Lip3K belonged to family I.3 lipase with a C-terminal secretion signal and showed the highest identity (93 %) to the lipase of Pseudomonas sp. MIS38. The presence of several newly identified conserved motifs in Est3K and Lip3K are suggested. Both Est3K and Lip3K exerted their maximal activity at pH 9.0 and 50 °C. The activity of Lip3K was significantly increased by the presence of 30 % methanol. The ability of the enzymes to retain activities in the presence of methanol and the substrates may offer a merit to the biotechnological applications of the enzymes such as transesterification. The activity and the thermostability of Lip3K were increased by Ca2+. Est3K and Lip3K preferred p-nitrophenyl butyrate (C4) and octanoate (C8), respectively, as the substrate and acted independently on the substrates with no synergistic effect.

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Acknowledgments

This study was supported by the Ministry of Environment of the Republic of Korea Grant No. 2008-05001-0033-0) and partially by the Suncheon Research Center for Natural Medicines, Republic of Korea.

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Authors and Affiliations

  1. Department of Pharmacy, Sunchon National University, Suncheon, 540-950, Republic of Korea

    Hee Jung Kim, Yu Seok Jeong, Hyun Woo Lee & Hoon Kim

  2. Suncheon Research Center for Natural Medicines, Suncheon, 540-950, Republic of Korea

    Won Kyeong Jung & Hoon Kim

  3. Department of Agricultural Chemistry, Sunchon National University, Suncheon, 540-950, Republic of Korea

    Sung Kyum Kim, Jungho Kim & Hoon Kim

  4. Department of Environmental Education, Sunchon National University, Suncheon, 540-950, Republic of Korea

    Hyung-Yeel Kahng

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  1. Hee Jung Kim
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Kim, H.J., Jeong, Y.S., Jung, W.K. et al. Characterization of Novel Family IV Esterase and Family I.3 Lipase from an Oil-Polluted Mud Flat Metagenome. Mol Biotechnol 57, 781–792 (2015). https://doi.org/10.1007/s12033-015-9871-4

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