Multilayer engineering of Saccharomyces cerevisiae for ovalicin secretion enables selective fungal removal in a skin microbiome mimic

초록

Engineered living cells can serve as functional biological platforms capable of interfacing with complex microenvironments through localized biochemical activity. Here, we established a Saccharomyces cerevisiae chassis engineered for the biosynthesis and extracellular delivery of ovalicin, an epoxide-bearing fungal metabolite with selective antifungal activity. Genome mining of Metarhizium anisopliae E6 using antiSMASH, combined with BLAST analysis based on fumagillin biosynthetic enzymes from Aspergillus fumigatus, enabled reconstruction of a putative ovalicin biosynthetic pathway in S. cerevisiae. Multi-layer engineering, including precursor flux redistribution, endoplasmic reticulum expansion, cytochrome P450 reductase optimization, and membrane transport control, progressively improved pathway performance and increased total ovalicin production to 5.8 mg/L. In particular, co-expression of the ABC transporter PDR5 with the lipid regulator OSH3 redirected metabolite partitioning and increased extracellular ovalicin from 0.2 mg/L to 3.5 mg/L. Direct application of the engineered yeast culture to a skin microbiome-mimicking consortium selectively depleted fungal members while preserving the viability of representative beneficial bacteria. Together, these results establish a yeast chassis that couples metabolite biosynthesis, export, and functional microbiome modulation, and provide an applied genetic and metabolic engineering strategy for constructing living microbial platforms with practical biological function.

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