Mechanism Behind the Programmed Biosynthesis of Heterotrimeric Fungal Depside Thielavin A

Thielavin A (1) is a fungal depside composed of one 3-methylorsellinic acid and two 3,5-dimethylorsellinic acid units. It displays diverse biological activities. However, the mechanism underlying the assembly of the heterotrimeric structure of 1 remains to be clarified. In this study, we identified the polyketide synthase (PKS) involved in the biosynthesis of 1. This PKS, designated as ThiA, possesses an unusual domain organization with the C-methyltransferase (MT) domain situated at the C-terminus following the thioesterase (TE) domain. Our findings indicated that the TE domain is solely responsible for two rounds of ester bond formation, along with subsequent chain hydrolysis. We identified a plausible mechanism for TE-catalyzed reactions and obtained insights into how a single PKS can selectively yield a specific heterotrimeric product. In particular, the tandem acyl carrier protein domains of ThiA are critical for programmed methylation by the MT domain. Overall, this study highlighted the occurrence of highly optimized domain-domain communication within ThiA for the selective synthesis of 1, which can advance our understanding of the programming rules of fungal PKSs. The polyketide synthase ThiA that forms the heterotrimeric fungal depside thielavin A (1) has been identified and characterized. The thioesterase (TE) domain of ThiA was found to perform two consecutive esterification reactions as well as subsequent hydrolysis, and a plausible reaction mechanism for the TE-catalyzed reactions has been proposed. Furthermore, this study provides insights into how ThiA selectivity yields the heterotrimeric product. image