Abstract
Monascus sp. is an important food microbial resource with the production of cholesterol-lowering agent lovastatin and other healthy metabolites. However, the mycotoxin citrinin naturally produced by Monascus sp. and the insufficient productivity of lovastatin limit its large-scale use in food industry. The aim of this paper is to modify a lovastatin-producing strain Monascus pilosus GN-01 through metabolic engineering to obtain a citrinin-free M. pilosus strain with higher yield of lovastatin. The citrinin synthesis regulator gene ctnR was firstly disrupted to obtain GN-02 without citrinin production. Based on that, the lovastatin biosynthesis genes (mokC, mokD, mokE, mokF, mokH, mokI, and LaeA) were, respectively, overexpressed, and pigment-regulatory gene (pigR) was knocked out to improve lovastatin production. The results indicated ctnR inactivation effectively disrupted the citrinin release by M. pilosus GN-01. The overexpression of lovastatin biosynthesis genes and pigR knockout could lead higher contents of lovastatin, of which pigR knockout strain achieved 76.60% increase in the yield of lovastatin compared to GN-02. These studies suggest that such multiplex metabolic pathway engineering in M. pilosus GN-01 is promising for high lovastatin production by a safe strain for application in Monascus-related food.
Key points
• Disruption of the regulator gene ctnR inhibited citrinin production of M. pilosus.
• Synchronous overexpression of biosynthesis gene enhanced lovastatin production.
• pigR knockout enhanced lovastatin of ΔctnR strain of M. pilosus.
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On reasonable request, the corresponding authors will provide the datasets created and/or analyzed during the current work.
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This work is financially supported by the grant from National Natural Science Foundation of China (32001634) and Natural Science Foundation of Fujian Province, China (2020J01488).
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HXK and GTL contributed equally to this work. HXK and GTL conducted experiments and wrote the manuscript. XXQ and LJ conceived and designed research. All authors have read and approved the final manuscript.
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Hong, X., Guo, T., Xu, X. et al. Multiplex metabolic pathway engineering of Monascus pilosus enhances lovastatin production. Appl Microbiol Biotechnol 107, 6541–6552 (2023). https://doi.org/10.1007/s00253-023-12747-2
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DOI: https://doi.org/10.1007/s00253-023-12747-2