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Expression of a lipase on the cell-surface of Escherichia coli using the OmpW anchoring motif and its application to enantioselective reactions

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Abstract

Microbial-surface display is the expression of proteins or peptides on the surface of cells by fusing an appropriate protein as an anchoring motif. Here, the outer membrane protein W (OmpW) was selected as a fusion partner for functional expression of Pseudomonas fluorescence SIK W1 lipase (TliA) on the cell-surface of Escherichia coli. Localization of the truncated OmpW-TliA fusion protein on the cell-surface was confirmed by immunoblotting and functional assay of lipase activity. Enantioselective hydrolysis of rac-phenylethyl butanoate by the displayed lipase resulted in optically active (R)-phenyl ethanol with 96 % enantiomeric excess and 44 % of conversion in 5 days. Thus, a small outer membrane protein OmpW, is a useful anchoring motif for displaying an active enzyme of ~50 kDa on the cell-surface and the surface-displayed lipase can be employed as an enantioselective biocatalyst in organic synthesis.

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Acknowledgments

This work was supported by the Basic Science Research Program (2010-0008826) through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (MEST). Further support from Priority Research Centers Program through the NRF funded by the MEST (2012-0006693) is appreciated.

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Authors and Affiliations

  1. Division of Drug Discovery Research, Korea Research Institute of Chemical Technology, P.O. Box 107, 141 Gajeong-ro, Yuseong-gu, Daejeon, 305-600, Republic of Korea

    Hyuk Lee & Seong Ho Kim

  2. Department of Environmental Engineering and Energy (Undergraduate Program) and Department of Energy Science and Technology (Graduate Program), Myongji University, San 38-2, Nam-dong, Cheoin-gu, Yongin-si, Gyeonggi-do, 449-728, Republic of Korea

    Si Jae Park

  3. Department of Biomolecular and Chemical Engineering, Dongyang University, 145 Dongyang-daero, Punggi-eup, Yeongju, Gyeongbuk, 750-711, Republic of Korea

    Mee-Jung Han

  4. Bio 1 Team, R&D Center, Samsung Petrochemical Co., Ltd., Nongseo-dong, Giheung-gu, Yongin-si, Gyeonggi-do, 446-712, Republic of Korea

    Gyeong Tae Eom

  5. Industrial Biochemicals Research Group, Research Center for Biobased Chemistry, Division of Convergence Chemistry, Korea Research Institute of Chemical Technology, P.O. Box 107, 141 Gajeong-ro, Yuseong-gu, Daejeon, 305-600, Republic of Korea

    Min-Jung Choi, Young Hoon Oh, Bong Keun Song & Seung Hwan Lee

Authors
  1. Hyuk Lee
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  2. Si Jae Park
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  3. Mee-Jung Han
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  6. Seong Ho Kim
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  7. Young Hoon Oh
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  9. Seung Hwan Lee
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Corresponding author

Correspondence to Seung Hwan Lee.

Additional information

Hyuk Lee and Si Jae Park are first co-authors and equally contributed to this study.

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Lee, H., Park, S.J., Han, MJ. et al. Expression of a lipase on the cell-surface of Escherichia coli using the OmpW anchoring motif and its application to enantioselective reactions. Biotechnol Lett 35, 1677–1683 (2013). https://doi.org/10.1007/s10529-013-1260-0

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  • DOI: https://doi.org/10.1007/s10529-013-1260-0

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