Abstract
Menaquinone-7 (MK-7), a highly valuable member of the vitamin K2 series, is an essential nutrient for humans. In this study, to develop engineered Escherichia coli strains for MK-7 production, heterogeneous heptaprenyl pyrophosphate synthetase (HepPPS) was introduced, and MK-7 production was first achieved in engineered E. coli by overexpression of Bacillus subtilis-derived HepPPS (BsHepPPS). Then, by optimizing the enzyme expression of the heterogenous mevalonic acid (MVA) pathway and the BsHepPPS, the titre of MK-7 increased to 2.3 μM, which was 22-fold higher than that of the original strain. The competitive pathways of MK-7 were further investigated by deletion of ubiCA or ispB. Finally, the scale-up fermentation of the engineered E. coli in a 5-L fermenter was studied under aerobic conditions using glucose, and 13.6 μM (8.8 mg/L) MK-7 was achieved. Additionally, metabolite analysis revealed a new bottleneck in the MK-7 pathway at ubiE, suggesting an avenue for further optimization. This report is the first to describe the metabolic engineering of MK-7 in E. coli, which provides a new perspective for MK-7 production.
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All data generated or analysed during this study are included in this published article [and its supplementary information files]. The authors agree that any materials newly described in the study are available to members of the scientific community for noncommercial purposes, if necessary, via an appropriate Materials Transfer Agreement between the interested parties.
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Acknowledgements
This study was funded by the National Key Research and Development Project of China (2018YFA0901400) and the National Natural Science Foundation of China Grant (31670051).
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This study was funded by the National Key Research and Development Project of China (2018YFA0901400) and the National Natural Science Foundation of China Grant (31670051).
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QG carried out the main work, collected and analysed the data, and drafted the manuscript. HC, WW and BL participated in the research. BL supervised the work and participated in data analysis and revised the manuscript. YT and JH participated in the conception and finalized the manuscript. All authors read and approved the final manuscript.
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Gao, Q., Chen, H., Wang, W. et al. Menaquinone-7 production in engineered Escherichia coli. World J Microbiol Biotechnol 36, 132 (2020). https://doi.org/10.1007/s11274-020-02880-9
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DOI: https://doi.org/10.1007/s11274-020-02880-9