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Exceptional thermal stability and organic solvent tolerance of an esterase expressed from a thermophilic host

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Abstract

A protein expression system recently developed for the thermophilic crenarchaeon Sulfolobus islandicus was employed to produce recombinant protein for EstA, a thermophilic esterase encoded in the same organism. Large amounts of protein were readily obtained by an affinity protein purification, giving SisEstA. Upon Escherichia coli expression, only the thioredoxin-tagged EstA recombinant protein was soluble. The fusion protein was then purified, and removing the protein tag yielded EcSisEstA. Both forms of the thermophilic EstA enzyme were characterized. We found that SisEstA formed dimer exclusively in solution, whereas EcSisEstA appeared solely as monomer. The former exhibited a stronger resistance to organic solvents than the latter in general, having a much higher temperature optimum (90°C vs. 65°C). More strikingly, SisEstA exhibited a half-life that was more than 32-fold longer than that of EcSisEstA at 90°C. This indicated that thermophilic enzymes yielded from homologous expression should be better biocatalysts than those obtained from mesophilic expression.

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Acknowledgment

We thank anonymous reviewers for the constructive suggestions for improving the manuscript. This research is supported by the State Key Laboratory of Agricultural Microbiology hosted by Huazhong Agricultural University and by a special grant from the same university, and by the Danish Free Research Council/FTP (grant no. 09-062932), Denmark to Q.S., and by a grant from the Science and Technology Department of Hubei Provincial Government, China to Y.L.

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Authors and Affiliations

  1. State Key Laboratory of Agricultural Microbiology, College of Life Science and Technology, Huazhong Agricultural University, Wuhan, 430070, People’s Republic of China

    Yuxia Mei, Nan Peng, Shumiao Zhao, Yongmei Hu, Huacai Wang & Yunxiang Liang

  2. Archaeal Centre, Department of Biology, University of Copenhagen, Ole Maaløes Vej 5, DK-2200, Copenhagen N, Denmark

    Nan Peng & Qunxin She

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  1. Yuxia Mei
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  2. Nan Peng
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  3. Shumiao Zhao
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  4. Yongmei Hu
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  5. Huacai Wang
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  6. Yunxiang Liang
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  7. Qunxin She
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Correspondence to Yunxiang Liang or Qunxin She.

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Mei, Y., Peng, N., Zhao, S. et al. Exceptional thermal stability and organic solvent tolerance of an esterase expressed from a thermophilic host. Appl Microbiol Biotechnol 93, 1965–1974 (2012). https://doi.org/10.1007/s00253-011-3504-z

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