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Enhancing menaquinone-7 biosynthesis through strengthening precursor supply and product secretion

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Abstract

Menaquinone-7 (MK-7) is an important class of vitamin K2 that is essential in human health and can prevent osteoporosis and cardiovascular disease. However, due to the complex synthesis pathway, the synthesis efficiency is low. The main objective of this study was to explore the effect of enhanced supply of precursors in Bacillus natto. Three precursors of pyruvate, shikimic acid, and sodium glutamate were chosen to investigate the effect of enhanced supply of precursors on MK-7 synthesis. Then, the optimal concentrations, different combinations, and different adding times were systematically studied, respectively. Results showed that the combination of shikimic acid and sodium glutamate could boost MK-7 production by 2 times, reaching 50 mg/L of MK-7 titer and 0.52 mg/(L·h) of MK-7 productivity. Furthermore, adding shikimic acid and sodium glutamate initially and feeding pyruvate at 48 h and 72 h increased MK-7 production to 58 mg/L. At the same time, the expression of the three related genes was also significantly upregulated. Subsequently, a new fermentation strategy combining the precursors enhancement and product secretion was proposed to enhance MK-7 yield and MK-7 productivity to 63 mg/L and 0.45 mg/(L·h). This study proposed a new fermentation regulation strategy for the enhancement of vitamin K2 biosynthesis.

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Data availability

The data that support the fndings of this study are available from the corresponding author upon reasonable request.

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Funding

This work was funded by the National Key Research and Development Program of China (No. 2019YFA0905700), the National Natural Science Foundation of China No. 21878151), the Natural Science Foundation of Jiangsu Province (BK20211535), the Jiangsu Synergetic Innovation Center for Advanced Bio-Manufacture (XTD2213), and Jiangsu Province “333” project (2022).

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Authors and Affiliations

  1. College of Biotechnology and Pharmaceutical Engineering, Nan**g Tech University, No. 30 South Puzhu Road, Nan**g, 211816, People’s Republic of China

    Pengchen Hu, Cheng Peng, Bei Zhang, Xuechao Hu, Ripon Baroi Milon & Lu**g Ren

  2. Shanghai JanStar Technology Development Co., Ltd., No. 1288, Huateng Road, Shanghai, People’s Republic of China

    Xuechao Hu

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  1. Pengchen Hu
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  2. Cheng Peng
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Contributions

Hu PC and Peng C designed the research, Ren LJ concept for this study and overall supervision of all experiments done, Zhang B and MRB conducted the research and analyzed the data, Hu XC helped with the statistics, Hu PC wrote the article.

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Correspondence to Lu**g Ren.

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Hu, P., Peng, C., Zhang, B. et al. Enhancing menaquinone-7 biosynthesis through strengthening precursor supply and product secretion. Bioprocess Biosyst Eng 47, 211–222 (2024). https://doi.org/10.1007/s00449-023-02955-5

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  • DOI: https://doi.org/10.1007/s00449-023-02955-5

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