摘要
2-Hydroxyisovalerate(2-HIV)is a value-added chemical that is widely applied in the synthesis of bioactive compounds and polymers.Here,we report an underexplored metabolic route for the de novo production of S-type 2-HIV(S-HIV)in Escherichia coli.In particular,we identified promiscuous activity of 4-hydroxymandelate syn-thase(HmaS)from Amycolatopsis orientalis towards the conversion of 2-keto-4-methyl-pentanoate(2-KMP,an immediate precursor for L-leucine)to S-HIV.Next,we designed a variant HmaS(S201F)with abolished activity for mandelate and 4-hydroxymandelate synthesis,thereby minimizing byproduct formation.Coupled with sys-tematic optimization of the L-leucine biosynthetic pathway,we achieved de novo production of S-HIV at 8.1 mM(0.95 g/L)in shake flasks and 33.9 mM(4.0 g/L)in 2-L fed-batch fermentation.In summary,this work represents the first time to realize the efficient synthesis of S-configuration 2-HIV in metabolically engineered E.coli.
基金
supported by the National Key R&D Program of China(grant no.:2024YFC3407000)
the National Natural Science Foundation of China(grant no.:32270087)
the Fundamental Research Funds for the Central Universities(grant no.:20720240120)
ZhenSheng Biotech.
