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A combination fermentation strategy for simultaneously increasing cellular NADP(H) level, biomass, and enzymatic activity of glufosinate dehydrogenase in Escherichia coli

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Abstract

Oxidoreductase is one of the most important biocatalysts for the synthesis of various chiral compounds. However, their whole-cell activity is frequently affected by an insufficient supply of expensive nicotinamide cofactors. This study aimed to overcome such shortcomings by developing a combination fermentation strategy for simultaneously increasing intracellular NADP(H) level, biomass, and glufosinate dehydrogenase activity in E. coli. The results showed that the feeding mode of NAD(H) synthesis precursor and lactose inducer had essential effects on the accumulation level of intracellular NADPH. Adding 40 mg L−1 of L-aspartic acid to the medium increased the intracellular NADP(H) concentration by 36.3%. Under the pH-stat feeding mode and adding 0.4 g L−1 h−1 lactose, the NADP(H) concentration, biomass, and GluDH activity in the 5-L fermenter reached 445.7 μmol L−1, 21.7 gDCW L−1, and 8569.3 U L−1, respectively. As far as we know, this is the highest reported activity of GluDH in the fermentation broth. Finally, the 5000-L fermenter was successfully scaled up to use this fermentation approach. The combination fermentation strategy might serve as a useful approach for the high-activity fermentation of other NADPH-dependent oxidoreductases.

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Data availability

The datasets used or analyzed during the current study are available from the corresponding author on reasonable request.

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Acknowledgements

This research/project is supported by the National Natural Science Foundation of China (NO. 21978268) and the National Key Research and Development Program of China (grant 2019YFA0905000).

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Authors and Affiliations

  1. Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou, 310014, China

    Shuping Zou, Jiawei Lu, Bing Zhang, Xia Li, Zhentao Jiang, Yaping Xue & Yuguo Zheng

  2. Engineering Research Center of Bioconversion and Biopurification of Ministry of Education, Zhejiang University of Technology, Hangzhou, 310014, China

    Shuping Zou, Jiawei Lu, Bing Zhang, Xia Li, Zhentao Jiang, Yaping Xue & Yuguo Zheng

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  1. Shuping Zou
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  2. Jiawei Lu
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  3. Bing Zhang
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  4. Xia Li
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  5. Zhentao Jiang
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  6. Yaping Xue
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Contributions

SZ: conceptualization, investigation, formal analysis, data curation, writing—original draft. JL: investigation, formal analysis, validation, writing—original draft. BZ: investigation, data curation. XL: investigation, data curation. ZJ: investigation, data curation. YX: writing—review and editing, funding acquisition. YZ: writing—review and editing, funding acquisition.

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Correspondence to Yaping Xue.

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Zou, S., Lu, J., Zhang, B. et al. A combination fermentation strategy for simultaneously increasing cellular NADP(H) level, biomass, and enzymatic activity of glufosinate dehydrogenase in Escherichia coli. Bioprocess Biosyst Eng 46, 867–878 (2023). https://doi.org/10.1007/s00449-023-02871-8

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