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High-Level Production of Recombinant Lipase by Fed-Batch Fermentation in Escherichia coli and Its Application in Biodiesel Synthesis from Waste Cooking Oils

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Abstract

The enzymatic production of biodiesel from waste cooking oils (WCOs) offers a green and sustainable solution for the liquid fuel manufacture as well as waste resource recovery. In present study, liquid lipase was used to simplify the catalysis process, thereby reducing biodiesel production costs. An engineered Escherichia coli expressing Geobacillus thermocatenulatus lipase 2 (GTL2) was screened at an enzyme activity of 6.96 U/mg, after evaluating the propagating stability of the recombinant plasmids exceeding 86.11%. Through the beneficial feeding strategy and effective pH control, high-level production of GTL2 by fed-batch fermentation was achieved with an enzyme activity of 434.32 U/mg, which was almost 62 times that of shake flask fermentation. In addition, liquid GTL2 was used to prepare fatty acid methyl esters (FAMEs) using WCOs. The effects of the reaction time, catalyst loading, temperature, and methanol-to-oil molar ratio on FAMEs production using WCOs were explored, and a maximum FAMEs yield of 96.62% was achieved under optimized conditions. These results indicate that liquid GTL2 is a promising biocatalyst for efficient utilization of WCOs in the synthesis of biodiesel and provide a novel enzymatic process for biodiesel reducing the cost of production.

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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 Natural Science Foundation of Guangdong Province of China (2020A1515010513), Program for Scientific Research Start-up Funds of Guangdong Ocean University (060302042110), National Natural Science Foundation of China (51903236 and 51606201), Projects of International Cooperation and Exchanges NSFC (51861145103), and Program of CAS Key Laboratory of Renewable Energy (E029010801).

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Authors and Affiliations

  1. College of Food Science and Technology, Guangdong Provincial Key Laboratory of Aquatic Product Processing and Safety, Guangdong Province Engineering Laboratory for Marine Biological Products, Guangdong Provincial Engineering Technology Research Center of Seafood, Key Laboratory of Advanced Processing of Aquatic Product of Guangdong Higher Education Institution, Guangdong Ocean University, Zhanjiang, 524088, China

    Jun Zhang

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

    Jun Zhang, Wen Luo, Zhiyuan Wang, Yiaoyan Chen, Junying Fu & Pengmei Lv

  3. Collaborative Innovation Center of Seafood Deep Processing, Dalian Polytechnic University, Dalian, 116034, China

    Jun Zhang

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

    Jingliang Xu

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  1. Jun Zhang
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  2. Wen Luo
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  3. Zhiyuan Wang
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Contributions

Jun Zhang: Conceptualization, methodology, investigation, software, data curation, writing—original draft, review and editing. Yiaoyan Chen and Jingliang Xu: Review and editing. Wen Luo, Zhiyuan Wang, Junying Fu, and Pengmei Lv: Data curation, funding acquisition, project administration, supervision, validation.

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Correspondence to Zhiyuan Wang or Pengmei Lv.

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Zhang, J., Luo, W., Wang, Z. et al. High-Level Production of Recombinant Lipase by Fed-Batch Fermentation in Escherichia coli and Its Application in Biodiesel Synthesis from Waste Cooking Oils. Appl Biochem Biotechnol 195, 432–450 (2023). https://doi.org/10.1007/s12010-022-04146-6

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