Enhancing Monacolin K and GABA Biosynthesis in <i>Monascus pilosus</i> via <i>GAD</i> Overexpression: Multi-Omics Elucidation of Regulatory Mechanisms
<i>Monascus</i> produces various bioactive compounds, including monacolin K (MK), γ-aminobutyric acid (GABA), and <i>Monascus</i> pigments (MPs). Studies have shown that overexpressing genes within the MK biosynthetic cluster significantly enhances MK production. Additionally...
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2025-07-01
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| author | Wenlan Mo Yiyang Cai Simei Huang Lishi Xiao Yanfang Ye Bin Yang Chan Zhang Zhiwei Huang |
| author_facet | Wenlan Mo Yiyang Cai Simei Huang Lishi Xiao Yanfang Ye Bin Yang Chan Zhang Zhiwei Huang |
| author_sort | Wenlan Mo |
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| description | <i>Monascus</i> produces various bioactive compounds, including monacolin K (MK), γ-aminobutyric acid (GABA), and <i>Monascus</i> pigments (MPs). Studies have shown that overexpressing genes within the MK biosynthetic cluster significantly enhances MK production. Additionally, MK synthesis in <i>Monascus</i> is regulated by other genes. Based on previous transcriptomic analyses conducted in our laboratory, a significant positive correlation was identified between the expression level of the <i>GAD</i> gene and MK production in <i>M. pilosus</i>. In this study, the <i>GAD</i> gene from <i>M. pilosus</i> was selected for overexpression, and a series of engineered <i>M. pilosus</i> strains were constructed. Among the 20 PCR-positive transformants obtained, 13 strains exhibited MK production increases of 12.84–52.50% compared to the parental strain, while 17 strains showed GABA production increases of 17.47–134.14%. To elucidate the molecular mechanisms underlying the enhanced production of MK and GABA, multi-omics analyses were performed. The results indicated that <i>GAD</i> overexpression likely promotes MK and GABA synthesis in <i>M. pilosus</i> by regulating key genes (e.g., <i>HPD</i>, <i>HGD</i>, and <i>FAH</i>) and metabolites (e.g., α-D-ribose-1-phosphate, β-alanine) involved in pathways such as tyrosine metabolism, phenylalanine metabolism, the pentose phosphate pathway, propanoate metabolism, and β-alanine metabolism. These findings provide theoretical insights into the regulatory mechanisms of MK and GABA biosynthesis in <i>Monascus</i> and suggest potential strategies for enhancing their production. |
| format | Article |
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| institution | Kabale University |
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| language | English |
| publishDate | 2025-07-01 |
| publisher | MDPI AG |
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| series | Journal of Fungi |
| spelling | doaj-art-4786b07b1852474caadf6c6ff3c7fea22025-08-20T03:35:28ZengMDPI AGJournal of Fungi2309-608X2025-07-0111750610.3390/jof11070506Enhancing Monacolin K and GABA Biosynthesis in <i>Monascus pilosus</i> via <i>GAD</i> Overexpression: Multi-Omics Elucidation of Regulatory MechanismsWenlan Mo0Yiyang Cai1Simei Huang2Lishi Xiao3Yanfang Ye4Bin Yang5Chan Zhang6Zhiwei Huang7Engineering Research Centre of Fujian-Taiwan Special Marine Food Processing and Nutrition (Ministry of Education), Fujian Agriculture and Forestry University, Fuzhou 350002, ChinaEngineering Research Centre of Fujian-Taiwan Special Marine Food Processing and Nutrition (Ministry of Education), Fujian Agriculture and Forestry University, Fuzhou 350002, ChinaEngineering Research Centre of Fujian-Taiwan Special Marine Food Processing and Nutrition (Ministry of Education), Fujian Agriculture and Forestry University, Fuzhou 350002, ChinaEngineering Research Centre of Fujian-Taiwan Special Marine Food Processing and Nutrition (Ministry of Education), Fujian Agriculture and Forestry University, Fuzhou 350002, ChinaEngineering Research Centre of Fujian-Taiwan Special Marine Food Processing and Nutrition (Ministry of Education), Fujian Agriculture and Forestry University, Fuzhou 350002, ChinaEngineering Research Centre of Fujian-Taiwan Special Marine Food Processing and Nutrition (Ministry of Education), Fujian Agriculture and Forestry University, Fuzhou 350002, ChinaSchool of Food and Health, Beijing Technology & Business University (BTBU), Beijing 100048, ChinaEngineering Research Centre of Fujian-Taiwan Special Marine Food Processing and Nutrition (Ministry of Education), Fujian Agriculture and Forestry University, Fuzhou 350002, China<i>Monascus</i> produces various bioactive compounds, including monacolin K (MK), γ-aminobutyric acid (GABA), and <i>Monascus</i> pigments (MPs). Studies have shown that overexpressing genes within the MK biosynthetic cluster significantly enhances MK production. Additionally, MK synthesis in <i>Monascus</i> is regulated by other genes. Based on previous transcriptomic analyses conducted in our laboratory, a significant positive correlation was identified between the expression level of the <i>GAD</i> gene and MK production in <i>M. pilosus</i>. In this study, the <i>GAD</i> gene from <i>M. pilosus</i> was selected for overexpression, and a series of engineered <i>M. pilosus</i> strains were constructed. Among the 20 PCR-positive transformants obtained, 13 strains exhibited MK production increases of 12.84–52.50% compared to the parental strain, while 17 strains showed GABA production increases of 17.47–134.14%. To elucidate the molecular mechanisms underlying the enhanced production of MK and GABA, multi-omics analyses were performed. The results indicated that <i>GAD</i> overexpression likely promotes MK and GABA synthesis in <i>M. pilosus</i> by regulating key genes (e.g., <i>HPD</i>, <i>HGD</i>, and <i>FAH</i>) and metabolites (e.g., α-D-ribose-1-phosphate, β-alanine) involved in pathways such as tyrosine metabolism, phenylalanine metabolism, the pentose phosphate pathway, propanoate metabolism, and β-alanine metabolism. These findings provide theoretical insights into the regulatory mechanisms of MK and GABA biosynthesis in <i>Monascus</i> and suggest potential strategies for enhancing their production.https://www.mdpi.com/2309-608X/11/7/506<i>Monascus pilosus</i><i>Glutamate decarboxylase</i> genemonacolin Kγ-aminobutyric acidoverexpression |
| spellingShingle | Wenlan Mo Yiyang Cai Simei Huang Lishi Xiao Yanfang Ye Bin Yang Chan Zhang Zhiwei Huang Enhancing Monacolin K and GABA Biosynthesis in <i>Monascus pilosus</i> via <i>GAD</i> Overexpression: Multi-Omics Elucidation of Regulatory Mechanisms Journal of Fungi <i>Monascus pilosus</i> <i>Glutamate decarboxylase</i> gene monacolin K γ-aminobutyric acid overexpression |
| title | Enhancing Monacolin K and GABA Biosynthesis in <i>Monascus pilosus</i> via <i>GAD</i> Overexpression: Multi-Omics Elucidation of Regulatory Mechanisms |
| title_full | Enhancing Monacolin K and GABA Biosynthesis in <i>Monascus pilosus</i> via <i>GAD</i> Overexpression: Multi-Omics Elucidation of Regulatory Mechanisms |
| title_fullStr | Enhancing Monacolin K and GABA Biosynthesis in <i>Monascus pilosus</i> via <i>GAD</i> Overexpression: Multi-Omics Elucidation of Regulatory Mechanisms |
| title_full_unstemmed | Enhancing Monacolin K and GABA Biosynthesis in <i>Monascus pilosus</i> via <i>GAD</i> Overexpression: Multi-Omics Elucidation of Regulatory Mechanisms |
| title_short | Enhancing Monacolin K and GABA Biosynthesis in <i>Monascus pilosus</i> via <i>GAD</i> Overexpression: Multi-Omics Elucidation of Regulatory Mechanisms |
| title_sort | enhancing monacolin k and gaba biosynthesis in i monascus pilosus i via i gad i overexpression multi omics elucidation of regulatory mechanisms |
| topic | <i>Monascus pilosus</i> <i>Glutamate decarboxylase</i> gene monacolin K γ-aminobutyric acid overexpression |
| url | https://www.mdpi.com/2309-608X/11/7/506 |
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