As an important functional monoterpene,α-terpineol can be generated through biotransformation.The transcription factor PdTP1 has been previously validated for its beneficial role inα-terpineol production via limonen...As an important functional monoterpene,α-terpineol can be generated through biotransformation.The transcription factor PdTP1 has been previously validated for its beneficial role inα-terpineol production via limonene biotransformation in Penicillium digitatum DSM 62840.However,the mechanistic basis of PdTP1-mediated transcriptional regulation remained elusive.In this study,DNA affinity purification sequencing was employed to identify the potential target genes of PdTP1.In combination with RNA sequencing data,a total of 981 genes were identified as the direct targets of PdTP1,participating in diverse metabolic pathways.Notably,cytochrome P450(CYP450)genes PDIDSM_08220 and PDIDSM_577403 were confirmed to be bound and activated by PdTP1 through yeast one-hybrid and dual luciferase report assays.Overexpression of PDIDSM_08220 and PDIDSM_577403 promoted limonene biotransformation and increasedα-terpineol production,and opposite results were observed after the silencing of these genes.These findings indicated that CYP450 genes PDIDSM_08220 and PDIDSM_577403 were regulated by transcription factor PdTP1,and played crucial roles in the biotransformation of limonene toα-terpineol,which will help us further understand the mechanism ofα-terpineol biosynthesis.展开更多
基金supported by the Science and Technology Project of China Tobacco Henan Industrial Co.,Ltd.(grant number AW2024035)the National Natural Science Foundation of China(grant number 32101868)+8 种基金the China Postdoctoral Science Foundation(grant number 2024M760795)the Postdoctoral Research Funding Project of Henan Province(grant number HN2025039)the Natural Science Research Project of Henan Educational Department(grant number 26ZX017)the Food Engineering Technology Research Center/Key Laboratory of Henan Province,Henan University of Technology(grant number GO202420)Cultivation Programme for Young Backbone Teachers in Henan University of Technology(grant number 21421235)Leading Talent Cultivation Project of Henan University of Technology(grant number 21421319)Henan University of Technology Postdoctoral Scientific Research Fund(grant number 21450084)the Key Research and Development Project of Henan Province(grant number 231111111800,251111113400)Cultivation Project of Tuoxin Team in Henan University of Technology(grant number 2024TXTD07).
文摘As an important functional monoterpene,α-terpineol can be generated through biotransformation.The transcription factor PdTP1 has been previously validated for its beneficial role inα-terpineol production via limonene biotransformation in Penicillium digitatum DSM 62840.However,the mechanistic basis of PdTP1-mediated transcriptional regulation remained elusive.In this study,DNA affinity purification sequencing was employed to identify the potential target genes of PdTP1.In combination with RNA sequencing data,a total of 981 genes were identified as the direct targets of PdTP1,participating in diverse metabolic pathways.Notably,cytochrome P450(CYP450)genes PDIDSM_08220 and PDIDSM_577403 were confirmed to be bound and activated by PdTP1 through yeast one-hybrid and dual luciferase report assays.Overexpression of PDIDSM_08220 and PDIDSM_577403 promoted limonene biotransformation and increasedα-terpineol production,and opposite results were observed after the silencing of these genes.These findings indicated that CYP450 genes PDIDSM_08220 and PDIDSM_577403 were regulated by transcription factor PdTP1,and played crucial roles in the biotransformation of limonene toα-terpineol,which will help us further understand the mechanism ofα-terpineol biosynthesis.
