Shu Wang (Class of ’16) and Huijue Lyu (Class of ’19)
In Vitro Reconstitution of the Remaining Steps in Ovothiol A Biosynthesis: C–S Lyase and Methyltransferase Reactions
Ovothiols are thiolhistidine derivatives. The first step of ovothiol biosynthesis is OvoA-catalyzed oxidative coupling between histidine and cysteine. In this report, the remaining steps of ovothiol A biosynthesis were reconstituted in vitro. ETA_14770 (OvoB) was reported as a PLP-dependent sulfoxide lyase, responsible for mercaptohistidine production. OvoA was found to be a bifunctional enzyme, which mediates both oxidative C–S bond formation and methylation of mercaptohistidine to afford ovothiol A. Besides reconstituting the whole biosynthetic pathway, two unique features proposed in the literature were also examined: a potential cysteine-recycling mechanism of the C–S lyase (OvoB) and the selectivity of the π-N methyltransferase.