A multi-enzyme cascade for efficient production of d-p-hydroxyphenylglycine from l-tyrosine

Abstract In this study, a four-enzyme cascade pathway was developed and reconstructed in vivo for the production of d - p -hydroxyphenylglycine (D-HPG), valuable intermediate used to produce ?-lactam antibiotics fine-chemical synthesis, from l -tyrosine. pathway, catalytic conversion 4-hydroxyphenylglyoxalate by meso -diaminopimelate dehydrogenase Corynebacterium glutamicum ( Cg DAPDH) identified as rate-limiting step, followed application mechanism-guided “conformation rotation” strategy decrease hydride-transfer distance (C6HDAP?C4NNADP) increase DAPDH activity. Introduction best variant generated protein engineering BC621/D120S/W144S/I169P with 5.32 ± 0.85 U·mg ?1 specific activity) into designed resulted D-HPG titer 42.69 g/L 50-g/L -tyrosine 24 h, 92.5% conversion, 71.5% isolated yield, > 99% enantiomeric excess 3-L fermenter. This provides an efficient enzymatic approach industrial cheap amino acids.