title: The c4h, tat, hppr and hppd genes prompted engineering of rosmarinic acid biosynthetic pathway in Salvia miltiorrhiza hairy root cultures.
abstract: Rational engineering to produce biologically active plant compounds has been greatly impeded by our poor understanding of the regulatory and metabolic pathways underlying the biosynthesis of these compounds. Here we capitalized on our previously described gene-to-metabolite network in order to engineer rosmarinic acid (RA) biosynthesis pathway for the production of beneficial RA and lithospermic acid B (LAB) in Salvia miltiorrhiza hairy root cultures. Results showed their production was greatly elevated by (1) overexpression of single gene, including cinnamic acid 4-hydroxylase (c4h), tyrosine aminotransferase (tat), and 4-hydroxyphenylpyruvate reductase (hppr), (2) overexpression of both tat and hppr, and (3) suppression of 4-hydroxyphenylpyruvate dioxygenase (hppd). Co-expression of tat/hppr produced the most abundant RA (906 mg/liter) and LAB (992 mg/liter), which were 4.3 and 3.2-fold more than in their wild-type (wt) counterparts respectively. And the value of RA concentration was also higher than that reported before, that produced by means of nutrient medium optimization or elicitor treatment. It is the first report of boosting RA and LAB biosynthesis through genetic manipulation, providing an effective approach for their large-scale commercial production by using hairy root culture systems as bioreactors.
authors.affiliation: Department of Pharmacy, Changzheng Hospital, Second Military Medical University, Shanghai, People's Republic of China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Xiao
authors.forename: Lei
authors.initials: L
authors.lastname: Zhang
authors.forename: Shouhong
authors.initials: S
authors.lastname: Gao
authors.forename: Saengking
authors.initials: S
authors.lastname: Saechao
authors.forename: Peng
authors.initials: P
authors.lastname: Di
authors.forename: Junfeng
authors.initials: J
authors.lastname: Chen
authors.forename: Wansheng
authors.initials: W
authors.lastname: Chen
mesh_terms: D053898:Biosynthetic Pathways / Q000235:genetics*; D002934:Cinnamates / Q000737:chemistry / Q000378:metabolism*; D053630:Depsides / Q000737:chemistry / Q000378:metabolism*; D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant / Q000235:genetics*; D060847:Metabolic Engineering / Q000379:methods*; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D018517:Plant Roots / Q000235:genetics / Q000254:growth & development*; D030821:Plants, Genetically Modified; D010957:Plasmids / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D015203:Reproducibility of Results; D027561:Salvia miltiorrhiza / Q000033:anatomy & histology / Q000235:genetics* / Q000254:growth & development; D046509:Tissue Culture Techniques / Q000379:methods*; D014170:Transformation, Genetic; D000097123:Rosmarinic Acid
chemical_list: D002934:Cinnamates; D053630:Depsides; D010940:Plant Proteins; D012333:RNA, Messenger; D000097123:Rosmarinic Acid