Medicarpin is an important bioactive compound with multiple medicinal activities,including anti-tumor,anti-osteoporosis,and anti-bacterial effects.Medicarpin is associated with pterocarpans derived from medicinal plan...Medicarpin is an important bioactive compound with multiple medicinal activities,including anti-tumor,anti-osteoporosis,and anti-bacterial effects.Medicarpin is associated with pterocarpans derived from medicinal plants,such as Sophora japonica,Glycyrrhiza uralensis Fisch.,and Glycyrrhiza glabra L.However,these medicinal plants contain only low amounts of medicarpin.Moreover,the planting area for medicarpin-producing plants is limited;consequently,the current medicarpin supply cannot meet the high demands of medicinal markets.In this study,eight key genes involved in medicarpin biosynthesis were identified using comparative transcriptome and bioinformatic analyses.In vitro and in vivo enzymatic reaction confirmed the catalytic functions of candidate enzymes responsible for the biosynthesis of medicarpin and medicarpin intermediates.Further engineering of these genes in Saccharomyces cerevisiae achieved the heterologous biosynthesis of medicarpin using liquiritigenin as a substrate,with a final medicarpin yield of 0.82±0.18 mg/L.By increasing the gene copy numbers of vestitone reductase(VR)and pterocarpan synthase(PTS),the final medicarpin yield was increased to 2.05±0.72 mg/L.This study provides a solid foundation for the economic and sustainable production of medicarpin through a synthetic biology strategy.展开更多
基金supported by the National Natural Science Foundation of China(81903526,81991523,82072240,and 32270192)the Open Project of Chinese Materia Medica First-Class Discipline of Nanjing University of Chinese Medicine(No.2020YLXK008 to W.L.)+3 种基金the Open Project of State Key Laboratory of Drug Research,Shanghai Institute of Materia Medica,Chinese Academy of Sciences(No.SIMM2205KF to W.L.)the Open Project of State Key Laboratory of Microbial Resources,Institute of Microbiology,Chinese Academy of Sciences(No.SKLMR-20220704 to W.L.)Jiangsu Graduate Research Innovation Program Project(KYCX22_2024 to CJ.L.)the Open Funding Project of the State Key Laboratory of Bioreactor Engineering,East China University of Science and Technology to W.L.,the Fok Ying Tung Education Foundation,and Jiangsu Specially-Appointed Professor Talent Program to W.L.
文摘Medicarpin is an important bioactive compound with multiple medicinal activities,including anti-tumor,anti-osteoporosis,and anti-bacterial effects.Medicarpin is associated with pterocarpans derived from medicinal plants,such as Sophora japonica,Glycyrrhiza uralensis Fisch.,and Glycyrrhiza glabra L.However,these medicinal plants contain only low amounts of medicarpin.Moreover,the planting area for medicarpin-producing plants is limited;consequently,the current medicarpin supply cannot meet the high demands of medicinal markets.In this study,eight key genes involved in medicarpin biosynthesis were identified using comparative transcriptome and bioinformatic analyses.In vitro and in vivo enzymatic reaction confirmed the catalytic functions of candidate enzymes responsible for the biosynthesis of medicarpin and medicarpin intermediates.Further engineering of these genes in Saccharomyces cerevisiae achieved the heterologous biosynthesis of medicarpin using liquiritigenin as a substrate,with a final medicarpin yield of 0.82±0.18 mg/L.By increasing the gene copy numbers of vestitone reductase(VR)and pterocarpan synthase(PTS),the final medicarpin yield was increased to 2.05±0.72 mg/L.This study provides a solid foundation for the economic and sustainable production of medicarpin through a synthetic biology strategy.
