High-efficiency expression of the thermophilic lipase from Geobacillus thermocatenulatus in Escherichia coli and its app
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ORIGINAL ARTICLE
High‑efficiency expression of the thermophilic lipase from Geobacillus thermocatenulatus in Escherichia coli and its application in the enzymatic hydrolysis of rapeseed oil Jun Zhang1,3 · Miao Tian1,3 · Pengmei Lv1 · Wen Luo1 · Zhiyuan Wang1 · Jingliang Xu1,2 · Zhongming Wang1 Received: 3 September 2020 / Accepted: 27 October 2020 © King Abdulaziz City for Science and Technology 2020
Abstract Long-chain fatty acids are widely used in food and chemical industries, and the enzymatic preparation of fatty acids is considered an environmentally friendly process. In the present study, long-chain fatty acids were prepared by the enzymatic hydrolysis of rapeseed oil with a genetically engineered lipase. Because thermophilic lipase has strong stability at higher temperatures, it was more suitable for the industrial production of long-chain fatty acids. Therefore, the thermophilic lipase BTL2 from Geobacillus thermocatenulatus was efficiently expressed in E. coli BL21(DE3) cells with an enzyme activity of 39.50 U/mg followed by gene codon optimisation. Experimental results showed that the recombinant lipase BTL2 exhibited excellent resistance to certain organic solvents (n-hexane, benzene, ethanol, and butanol). The metal cation Ca2+ and the non-ionic surfactant Triton-100X enhanced enzyme activity by 7.36% and 56.21% respectively. Moreover, the acid value of the liberated long-chain fatty acids by hydrolysing rapeseed oil was approximately 161.64 mg KOH/g at 50 °C in 24 h, the hydrolytic conversion rate was 91.45%, and the productivity was approximately 6.735 mg KOH/g h. These results suggested that the recombinant lipase BTL2 has excellent hydrolytic performance for rapeseed oil and showed great potential for the enzymatic preparation of long-chain fatty acids. Keywords Thermophilic lipase · Escherichia coli · Enzymatic hydrolysis · Rapeseed oil
Introduction
Electronic supplementary material The online version of this article (https://doi.org/10.1007/s13205-020-02517-6) contains supplementary material, which is available to authorized users. * 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
School of Chemical Engineering, Zhengzhou University, Zhengzhou 450001, China
3
University of China Academy of Sciences, Beijing 100049, China
Lipase (EC 3.1.1.3) is any enzyme that catalyses the hydrolysis of triglycerides. Its diverse array of uses can be seen in many fields ranging from food and medicine to detergents and biofuels (Sarmah et al. 2018). In recent years, thermophilic lipases have gained increasing attention because of its stability at elevated temperatures (He et al. 2016). Lipase 2 from Geobacillus thermocatenulatus (BTL2) belongs to the lipase family 1.5, which is a typical thermophilic lipase with excellent catalytic properties such as
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