De novo production of aromatic m-cresol in Saccharomyces cerevisiae mediated by heterologous polyketide synthases combined with a 6-methylsalicylic acid decarboxylase
- PMID: 31193192
- PMCID: PMC6520567
- DOI: 10.1016/j.mec.2019.e00093
De novo production of aromatic m-cresol in Saccharomyces cerevisiae mediated by heterologous polyketide synthases combined with a 6-methylsalicylic acid decarboxylase
Abstract
As a flavor and platform chemical, m-cresol (3-methylphenol) is a valuable industrial compound that currently is mainly synthesized by chemical methods from fossil resources. In this study, we present the first biotechnological de novo production of m-cresol from sugar in complex yeast extract-peptone medium with the yeast Saccharomyces cerevisiae. A heterologous pathway based on the decarboxylation of the polyketide 6-methylsalicylic acid (6-MSA) was introduced into a CEN.PK yeast strain. For synthesis of 6-MSA, expression of different variants of 6-MSA synthases (MSASs) were compared. Overexpression of codon-optimized MSAS from Penicillium patulum together with activating phosphopantetheinyl transferase npgA from Aspergillus nidulans resulted in up to 367 mg/L 6-MSA production. Additional genomic integration of the genes had a strongly promoting effect and 6-MSA titers reached more than 2 g/L. Simultaneous expression of 6-MSA decarboxylase patG from A. clavatus led to the complete conversion of 6-MSA and production of up to 589 mg/L m-cresol. As addition of 450-750 mg/L m-cresol to yeast cultures nearly completely inhibited growth our data suggest that the toxicity of m-cresol might be the limiting factor for higher production titers.
Keywords: 6-Methylsalicylic acid decarboxylase; 6-Methylsalicylic acid synthase; 6-methylsalicylic acid decarboxylase, PatG; 6-methylsalicylic acid synthase, MSAS; 6-methylsalicylic acid, 6-MSA; Acyl carrier protein, ACP; Acyltransferase, AT; Codon-optimization; Polyketide synthase; Saccharomyces cerevisiae; ketoreductase, KR; ketosynthase, KS; m-Cresol; optical density, OD; phosphopantetheinyl transferase, PPT; polyketide synthase, PKS; thioester hydrolase, TH.
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