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Sugar Ester Synthesis by Thermostable Lipase from Streptomyces thermocarboxydus ME168

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Abstract

The extracellular lipase from Streptomyces thermocarboxydus ME168 was purified to 9.5-fold with 20% yield, following concentration by acetone precipitation, ion exchange chromatography (Resource Q) and gel filtration chromatography (Superdex 200), respectively. The purified enzyme had an apparent molecular mass of 21 kDa by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The N-terminal sequence of the lipase was ASDFDDQILG and was different from most other reported lipase. The enzyme showed maximum activity at 50 °C with the half-life of 180 min at 65 °C. It showed high stability at a broad pH range of 5.5–9.5 and was thermostable at the temperature range of 25–60 °C. The K m and V max were 0.28 mM and 1,428 U/mg, respectively, using p-nitrophenyl palmitate as substrate. It was active toward p-nitrophenyl ester with medium to long acyl chain (C8–C16). Lipase activity was inhibited by Zn2+, dithiothreitol (DTT), EDTA and some organic solvents, e.g., ethanol, acetone, dioxane, acetronitrile, tert-butanol and pyridine. Immobilized crude lipase of S. thermocarboxydus ME168 on celite could be used to synthesize sugar esters from glucose and vinyl acetate, vinyl butyrate or vinyl caproate in tert-butanol:pyridine (55:45 v/v) at 45 °C with conversion yields of 93, 67 and 55%, respectively.

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Acknowledgments

The financial support by the Royal Golden Jubilee Program (RGJ, Thailand) to Thanongsak Chaiyaso was highly appreciated. The partial support from German Academic Exchange Service (DAAD, Germany) was also gratefully acknowledged.

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Authors and Affiliations

  1. Division of Biotechnology, Faculty of Agro-Industry, Chiang Mai University, Chiang Mai, 50100, Thailand

    Phisit Seesuriyachan & Thanongsak Chaiyaso

  2. Department of Industrial Biotechnology, Faculty of Agro-Industry, Prince of Songkla University, Hat Yai, 90112, Thailand

    Aran H-Kittikun & Poonsuk Prasertsan

  3. Department of Microbiology and Bioprocess Technology, Institute of Biochemistry, University of Leipzig, 04103, Leipzig, Germany

    Wolfgang Zimmermann

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  1. Aran H-Kittikun
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  2. Poonsuk Prasertsan
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  4. Phisit Seesuriyachan
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  5. Thanongsak Chaiyaso
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Correspondence to Aran H-Kittikun.

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H-Kittikun, A., Prasertsan, P., Zimmermann, W. et al. Sugar Ester Synthesis by Thermostable Lipase from Streptomyces thermocarboxydus ME168. Appl Biochem Biotechnol 166, 1969–1982 (2012). https://doi.org/10.1007/s12010-012-9624-9

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