Synthetic biology identifies the minimal gene set required for paclitaxel biosynthesis in a plant chassis  

作　　者：Youjun Zhang Lorenz Wiese Hao Fang Saleh Alseekh Leonardo Perez de Souza Federico Scossa John Molloy Mathias Christmann Alisdair R.Fernie 

机构地区：[1]Max-Planck-Institut fur Molekulare Pflanzenphysiologie,Am Muhlenberg 1,14476 Potsdam-Golm,Germany [2]Center of Plant Systems Biology and Biotechnology,4000 Plovdiv,Bulgaria [3]Institute of Chemistry and Biochemistry,Freie Universitat Berlin,TakustraBe 3,14195 Berlin,Germany [4]Department of Biomolecular Systems,Max Planck Institute of Colloids and Interfaces,14476 Potsdam,Germany [5]Research Center for Genomics and Bioinformatics(CREA-GB),Via Ardeatina 546,00178 Rome,Italy

出　　处：《Molecular Plant》2023年第12期1951-1961,共11页分子植物（英文版）

基　　金：the Max Planck Society(Y.Z-.,S.A.,L.P.d.S.,F.S.,and A.R.F.),and Y.Z.,S.A.,and A.R.F.acknowledge the European Union's Horizon 2020 research and innovation programme,project PlantaSYST(SGA-CSA No.739582 under FPA No.664620);the BG05M2OP001-1.003-001-C01 project,financed by the European Regional Development Fund through the Bulgarian"Science and Education for Smart Growth"Operational Programme.J.J.M.thanks the Fonds der Chemischen Industrie,FCI for funding,H.F.thanks the Chinese Scholarship Councilfor funding.

摘　　要：The diterpenoid paclitaxel(Taxol)is a chemotherapy medication widely used as a first-line treatment against several types of solid cancers.The supply of paclitaxel from natural sources is limited.However,missing knowledge about the genes involved in several specific metabolic steps of paclitaxel biosynthesis has rendered it difficult to engineer the full pathway.In this study,we used a combination of transcriptomics,cell biology,metabolomics,and pathway reconstitution to identify the complete gene set required for the heterologous production of paclitaxel.We identified the missing steps from the current model of paclitaxel biosynthesis and confirmed the activity of most of the missing enzymes via heterologous expression in Nicotiana benthamiana.Notably,we identified a new C4β-C20 epoxidase that could overcome the first bottleneck of metabolic engineering.We used both previously characterized and newly identified oxomutases/epoxidases,taxane 1β-hydroxylase,taxane 9aα-hydroxylase,taxane 9α-dioxygenase,and phenylalanine-CoA ligase,to successfully biosynthesize the key intermediate baccatin Ill and to convert baccatin Ill into paclitaxel in N.benthamiana.In combination,these approaches establisha metabolic route to taxoidbiosynthesis and provide insights into the unique chemistry that plants use to generate complex bioactive metabolites.

关 键 词：paclitaxel biosynthesis synthetic biology baccatin Ill biosynthesis TAXUS 

分 类 号：Q94[生物学—植物学]