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Heterologous expression of lipase in Escherichia coli is limited by folding and disulfide bond formation

  • Applied Genetics and Molecular Biotechnology
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Abstract

Functional expression of lipase B from Pseudozyma antarctica (PalB) in the cytoplasm of Escherichia coli BL21(DE3) and its mutant derivative Origami B(DE3) was explored. Coexpression of DsbA was found to be effective in enhancing PalB expression. The improvement was particularly pronounced with Origami B(DE3) as a host, suggesting that both folding and disulfide bond formation may be major factors limiting PalB expression. Fusion tag technique was also explored by constructing several PalB fusions for the evaluation of their expression performance. While the solubility was enhanced for most PalB fusions, only the DsbA tag was effective in boosting PalB activity, possibly by both enhanced solubility and correct disulfide bond formation. Our results suggest that PalB activity is closely associated with correct disulfide bond formation, and increased solubilization by PalB fusions does not necessarily result in activity enhancement.

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Acknowledgments

This study is supported by the Natural Sciences and Engineering Research Council (NSERC) of Canada and the Canada Research Chair (CRC) program. We thank Deb K. Chatterjee, Didier Busso, Nikolay K. Tzvetkov, and EMBL for providing various vectors in this study.

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Authors and Affiliations

  1. Department of Chemical Engineering, University of Waterloo, 200 University Avenue West, Waterloo, Ontario, Canada, N2L 3G1

    Yali Xu, Amrita Yasin, Raymond Tang, Jeno M. Scharer, Murray Moo-Young & C. Perry Chou

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  1. Yali Xu
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  2. Amrita Yasin
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  3. Raymond Tang
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  4. Jeno M. Scharer
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  5. Murray Moo-Young
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  6. C. Perry Chou
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Correspondence to C. Perry Chou.

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Xu, Y., Yasin, A., Tang, R. et al. Heterologous expression of lipase in Escherichia coli is limited by folding and disulfide bond formation. Appl Microbiol Biotechnol 81, 79–87 (2008). https://doi.org/10.1007/s00253-008-1644-6

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  • DOI: https://doi.org/10.1007/s00253-008-1644-6

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