Regulation of MareA Gene on Monascus Growth and Metabolism Under Different Nitrogen Sources.

Abstract

Monascus is a widely used natural microorganism in our country, which can produce useful secondary metabolites. Studies have shown that the nitrogen source directly affects the growth, reproduction, and secondary metabolites of Monascus. As a global transcriptional regulator of nitrogen metabolism, MareA gene is involved in the regulation of secondary metabolism. In this study, we found the MareA gene that is highly homologous to the AreA gene sequence, and used MareA to obtain ΔMareA and OE-MareA. Three strains were cultured with glutamine, urea, NaNO₃, and (NH₄)₂SO₄ nitrogen sources. The Monascus pigments and related genes were analyzed by solid-state fermentation under different nitrogen sources. The results showed that the pigment production of the ΔMareA decreased, but the OE-MareA did the opposite. The secondary metabolites of the three strains were analyzed by HPLC and expression level of pigment biosytnthesis gene was determined by RT-qPCR. The relative expression levels of four key Monascus pigment genes in ΔMareA were significantly upregulated in mppE gene, but downregulated in MpPKS5, mppG, and mppD genes. Monascus pigment genes were increased in OE-MareA. In terms of growth regulation, the expression of VosA and LaeA genes was significantly reduced in ΔMareA, while OE-MareA significantly promoted the expression of GprD genes. The pigment production and gene expression in ΔMareA were significantly lower than that of C100, while the opposite was true of OE-MareA when NaNO₃ was added to the culture medium. In conclusion, MareA gene had different regulatory effects on Monascus growth and pigments metabolism under different nitrogen sources.