Co-Expression of a Thermally Stable and Methanol-Resistant Lipase and Its Chaperone from Burkholderia cepacia G63 in Esc
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Co-Expression of a Thermally Stable and Methanol-Resistant Lipase and Its Chaperone from Burkholderia cepacia G63 in Escherichia coli Jun Zhang 1,2 & Miao Tian 1,2 & Xiaoyan Chen 1,2 & Pengmei Lv 1 & Wen Luo 1 & Zhiyuan Wang 1 & Jingliang Xu 1,3 & Zhongming Wang 1 Received: 14 September 2020 / Accepted: 8 November 2020/ # Springer Science+Business Media, LLC, part of Springer Nature 2020
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. Keywords Lipase . Lipase chaperone . Co-expression . Escherichia coli . Biodiesel
* Pengmei Lv [email protected] * Jingliang Xu [email protected]
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
2
University of China Academy of Sciences, Beijing 100049, China
3
School of Chemical Engineering, Zhengzhou University, Zhengzhou 450001, China
Applied Biochemistry and Biotechnology
Introduction Biodiesel is mainly produced by chemical methods, but it has caused excessive environmental pollution and energy consumption [13, 25]. However, the transesterification of short-chain alcohols via lipase to synthesize biodiesel is a green production method. This method has characteristics of less by-products, low energy consumption, and environmental friendliness, and is expected to be applied in industrial biodiesel production. Lipase is an important industrial biocatalyst, widely used in bioenergy and pharmaceutical fields [19]. Moreover, most of lipases derived from Burkholderia cepacia are very suitable for biodiesel production due to its favorable thermal stability, high methanol tolerance, and broad adaptability to oil source [27], especially the lipase from B. cepacia G63 strain, such as preserved enzymatic activity for long time in certain organic solvents or at high tem
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