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Polycationic amino acid tags enhance soluble expression of Candida antarctica lipase B in recombinant Escherichia coli

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Abstract

Lipase (EC 3.1.1.3) is a popular enzyme used as an ingredient in detergents and biocatalyst in many biochemical reactions. Lipase is usually expressed in Escherichia coli as an inactive inclusion body and at a low level. In this study, Candida antarctica lipase B (CalB) was fused with various polycationic amino acid tags and expressed in E. coli in order to increase a soluble expression level. By induction with 1.0 mM IPTG, the authentic and fused CalBs were expressed at 27–56% of total protein. The 10-arginine and 10-lysine tags fused at the C-terminal of CalB significantly increased the solubility of CalB by five- to ninefold, relative to the case of the authentic CalB expressed in a recombinant E. coli Origami 2TM (DE3) strain. Among a series of the C-terminal poly-arginine tags, the recombinant CalB combined with the 10-arginine tag (CalB-R10) possessed the highest lipase specific activity of 9.5 ± 0.03 U/mg protein, corresponding to a fourfold enhancement compared with the authentic CalB.

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Acknowledgments

This research was supported by Basic Science Research Program (2010-0015928) through the National Research Foundation of Korea (NRF), and the Advanced Biomass R&D Center (ABC) of Korea Grant (2010-0029799) funded by the Ministry of Education, Science and Technology, and in part by the new faculty research program 2009 of Kookmin University in Korea.

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Authors and Affiliations

  1. Department of Agricultural Biotechnology and Center for Agricultural Biomaterials, Seoul National University, Seoul, 151-921, Korea

    Hyun-Jung Jung, Sun-Ki Kim, Won-Ki Min & Jin-Ho Seo

  2. Division of Forest Bioenergy, Korea Forest Research Institute, Seoul, 130-712, Korea

    Sung-Suk Lee

  3. Department of Biological and Chemical Engineering, Hongik University, Jochiwon, Chungnam, 339-701, Korea

    Kyungmoon Park

  4. Department of Advanced Fermentation Fusion Science and Technology, Kookmin University, Seoul, 136-702, Korea

    Yong-Cheol Park

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  1. Hyun-Jung Jung
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  2. Sun-Ki Kim
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  3. Won-Ki Min
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  4. Sung-Suk Lee
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  5. Kyungmoon Park
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  6. Yong-Cheol Park
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  7. Jin-Ho Seo
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Correspondence to Yong-Cheol Park or Jin-Ho Seo.

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Jung, HJ., Kim, SK., Min, WK. et al. Polycationic amino acid tags enhance soluble expression of Candida antarctica lipase B in recombinant Escherichia coli . Bioprocess Biosyst Eng 34, 833–839 (2011). https://doi.org/10.1007/s00449-011-0533-z

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  • DOI: https://doi.org/10.1007/s00449-011-0533-z

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