Fig. 2
From: CRISPRi-mediated metabolic engineering of E. coli for O-methylated anthocyanin production
Modular ortholog screen identifies several AOMTs that are capable of producing peonidin 3-O-glucoside (P3G) in E. coli. a The upstream module, encoded on low-copy ePathBrick vector pACM4, produces cyanidin 3-O-glucoside (C3G) from (+)-catechin and endogenous UDP-glucose, b all AOMT orthologs were cloned into compatible high-copy ePathBrick vector pETM6 to facilitate rapid screening of downstream modules following combinatorial co-transformation with upstream module. The downstream module converts C3G and endogenous S-adenosylmethionine (SAM) to P3G, c ortholog screen reveals VvAOMT1 and CkmOMT2 as best P3G (blue) producers overall (top), while VvAOMT1 achieved best conversion of the final C3G to P3G step as demonstrated by the high final P3G:C3G titer ratio (bottom)