Heterologous expression of a polyketide synthase ACRTS2 in Aspregillus oryzae produces host selective ACR-toxins: Co-production of minor metabolites.

Previously, we succeeded to produce the core structure of the host-selective ACR-toxin (1) on brown leaf spot on rough lemon when the polyketide synthase ACRTS2 gene was heterologously expressed in Aspergillus oryzae (AO). To confirm the production of 1 in AO, the detection limit and suppressing decarboxylation were improved, and these efforts led us to conclude the direct production of 1 instead of its decarboxylation product. During this examination, minor ACR-toxin-related metabolites were found. Their structure determination enabled us to propose a decarboxylation mechanism and novel branching route forming byproducts from the coupling of the dihydropyrone moiety of 1 with the acetaldehyde and kojic acid abundant in AO. The involvement of putative cyclase ACRTS3 in the chain release of linear polyketide was excluded by the co-expression analysis of ACRTS2 and ACRTS3. Taken together, we concluded the production of 1 in AO is solely responsible for ACRTS2.