单羧酸转运蛋白MpMch2调控紫色红曲霉的性发育及代谢产物的产生

Monocarboxylate transporter MpMch2 regulates the sexual development and metabolite production of Monascus purpureus

乳酸、丙酮酸、酮体等单羧酸盐在生物体的代谢活动中发挥重要作用。MpMch2作为单羧酸转运蛋白,主要负责单羧酸盐的跨膜转运,维持葡萄糖代谢平衡等。【目的】在紫色红曲霉中对单羧酸转运蛋白MpMch2进行功能分析。【方法】以紫色红曲霉Mp-21为出发菌株,通过构建敲除载体,利用同源重组的方式将潮霉素基因替换MpMch2得到缺失株∆MpMch2。观察Mp-21和∆MpMch2在不同培养基上菌落形态、显微形态,测定其红曲色素、γ-氨基丁酸产量、分生孢子、子囊孢子产量等;利用RT-qPCR检测分生孢子、γ-氨基酸丁酸相关基因表达量。【结果】在不同培养基上野生型和∆MpMch2在菌落形态上无显著差异,该基因敲除后菌株分生孢子、子囊孢子产量下降,红曲色素、γ-氨基丁酸产量及相关基因表达量下降。【结论】表明MpMch2基因正调控分生孢子和子囊孢子发育及红曲色素、γ-氨基丁酸产量。

Monocarboxylates such as lactate,pyruvate,and ketone bodies play an important role in the metabolic activities of organisms.As a monocarboxylate transporter,MpMch2 is mainly responsible for the transmembrane transport of monocarboxylates and the maintenance of glucose metabolism balance.[Objective]To analyze the functions of MpMch2 in Monascus purpureus.[Methods]The MpMch2 in M.purpureus Mp-21 was replaced with the hygromycin gene by homologous recombination to construct the deletion strain∆MpMch2.The colony and cell morphology of Mp-21 and∆MpMch2 on different media was observed,and the yield of monascus pigment,γ-aminobutyric acid,conidia and ascospores were determined.The expression levels of genes related to conidia andγ-aminobutyric acid were determined by RT-qPCR.[Results]There was no significant difference in the colony morphology between the wild type and∆MpMch2 on different media.After knockout of MpMch2,the yields of conidia,ascospores,Monascus pigments,andγ-aminobutyric acid decreased,and the expression levels of related genes were down-regulated.[Conclusion]MpMch2 positively regulated the development of conidia and ascospores and the production of Monascus pigments andγ-aminobutyric acid.