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.
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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), the National Natural Science Foundation of China (No. 51903236) and the Projects of International Cooperation and Exchanges NSFC (No. 51861145103).
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Conceptualization, Methodology, Software, Investigation, Writing—original draft: ZJ; Validation, Formal analysis, Visualization, Software: TM; Supervision, Data curation: LW and WZY; Resources, Writing—review & editing, Supervision, Data curation: LPM, XJL and WZM.
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Zhang, J., Tian, M., Lv, P. et al. High-efficiency expression of the thermophilic lipase from Geobacillus thermocatenulatus in Escherichia coli and its application in the enzymatic hydrolysis of rapeseed oil. 3 Biotech 10, 523 (2020). https://doi.org/10.1007/s13205-020-02517-6
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DOI: https://doi.org/10.1007/s13205-020-02517-6