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Co-Expression of a Thermally Stable and Methanol-Resistant Lipase and Its Chaperone from Burkholderia cepacia G63 in Escherichia coli

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Abstract

Biodiesel biosynthesis with enzymatic transesterification is considered green, sustainable, and environmentally friendly method. Lipase from Burkholderia cepacia G63 has excellent catalytic properties in biodiesel production. Lipase chaperones promote secretion and folding of enzymes, thereby enhancing enzymatic activity. In the current study, heterologous co-expression of lipase (lipA) and chaperone (lipB) was achieved in Escherichia coli through codon optimization. The enzymatic activity of purified and renatured lipAB was 2080.23 ± 19.18 U/g at 50 °C and pH 8.0. Moreover, lipAB showed increased resistance to pH and temperature changes, and lipAB retained stable catalytic properties after treatment with metal ions, organic solvents, and surfactants, namely Mg2+, methanol, and Triton-100X. Besides, using recombinant lipase lipAB as catalysts, biodiesel was synthesized using rapeseed oil under 50 °C for 72 h with a yield of 90.23%. Thus, the current study confirmed that co-expression of lipase and its chaperone is an effective strategy to enhance enzyme activity and improve the biochemical profile, meanwhile, showing that lipAB is a promising biocatalyst for biodiesel production.

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Data Availability

The raw/processed data required to reproduce these findings cannot be shared at this time as the data also forms part of an ongoing study.

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Funding

This work was supported by the Guangdong Special Support Program (No. 2017TX04Z109), the National Natural Science Foundation of China (No. 51606201), Science and Technology Planning Project of Guangdong Province (No. 2016A010104008), the Natural Science Foundation of Guangdong Province (No. 2017A010104010), the National Key Research and Development Program of China (No. 2019YFB1504003), and the Projects of International Cooperation and Exchanges NSFC (No. 51861145103).

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Authors and Affiliations

  1. Guangzhou Institute of Energy Conversion, Chinese Academy of Sciences, CAS Key Laboratory of Renewable Energy, Guangdong Provincial Key Laboratory of New and Renewable Energy Research and Development, Guangzhou, 510640, China

    Jun Zhang, Miao Tian, Xiaoyan Chen, Pengmei Lv, Wen Luo, Zhiyuan Wang, Jingliang Xu & Zhongming Wang

  2. University of China Academy of Sciences, Beijing, 100049, China

    Jun Zhang, Miao Tian & Xiaoyan Chen

  3. School of Chemical Engineering, Zhengzhou University, Zhengzhou, 450001, China

    Jingliang Xu

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  1. Jun Zhang
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  2. Miao Tian
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  4. Pengmei Lv
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Contributions

J. Z analyzed data, wrote, reviewed, and edited the manuscript. J. Z, P M. L, and J L. X conceived and designed the research. M. T and X Y. C contributed new reagents or analytical tools, and reviewed and edited the manuscript. W. L and Z Y. W conducted experiments. P M. L, J L. X, and Z M. W reviewed and edited the manuscript. All authors read and approved the manuscript.

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Correspondence to Pengmei Lv or Jingliang Xu.

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Zhang, J., Tian, M., Chen, X. et al. Co-Expression of a Thermally Stable and Methanol-Resistant Lipase and Its Chaperone from Burkholderia cepacia G63 in Escherichia coli. Appl Biochem Biotechnol 193, 717–729 (2021). https://doi.org/10.1007/s12010-020-03453-0

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