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Here we present an inexpensive production method for 3-ethylphenol (3-EP) and 3-propylphenol (3-PP) by microbial fermentation of sugars. Heterologous expression in the yeast <jats:italic>Saccharomyces cerevisiae<\/jats:italic> of phosphopantetheinyltransferase-activated 6-methylsalicylic acid (6-MSA) synthase (MSAS) and 6-MSA decarboxylase converted acetyl-CoA as a priming unit via 6-MSA into 3-methylphenol (3-MP). We exploited the substrate promiscuity of MSAS to utilize propionyl-CoA and butyryl-CoA as alternative priming units and the substrate promiscuity of 6-MSA decarboxylase to produce 3-EP and 3-PP in yeast fermentations. Increasing the formation of propionyl-CoA by expression of a bacterial propionyl-CoA synthetase, feeding of propionate and blocking propionyl-CoA degradation led to the production of up to 12.5\u2009mg\/L 3-EP. Introduction of a heterologous \u2018reverse \u00df-oxidation\u2019 pathway provided enough butyryl-CoA for the production of 3-PP, reaching titers of up to 2.6\u2009mg\/L. As the concentrations of 3-alkylphenols are close to the range of the concentrations deployed in tsetse fly traps, the yeast broths might become promising and inexpensive sources for attractants, producible on site by rural communities in Africa.<\/jats:p>","DOI":"10.1038\/s41598-020-66997-5","type":"journal-article","created":{"date-parts":[[2020,6,19]],"date-time":"2020-06-19T10:03:27Z","timestamp":1592561007000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Substrate promiscuity of polyketide synthase enables production of tsetse fly attractants 3-ethylphenol and 3-propylphenol by engineering precursor supply in yeast"],"prefix":"10.1038","volume":"10","author":[{"given":"Julia","family":"Hitschler","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7269-0667","authenticated-orcid":false,"given":"Martin","family":"Grininger","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Eckhard","family":"Boles","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2020,6,19]]},"reference":[{"key":"66997_CR1","doi-asserted-by":"publisher","first-page":"281","DOI":"10.1017\/S0007485300013043","volume":"78","author":"E Bursell","year":"1988","unstructured":"Bursell, E. et al. 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