Production of the polyketide 6-MSA in yeast engineered for increased malonyl-CoA supply

• Published in: Metabolic engineering 2008-09, Vol.10 (5), p.246-254
• Main Authors: Wattanachaisaereekul, Songsak, Lantz, Anna Eliasson, Nielsen, Michael Lynge, Nielsen, Jens
• Format: Article
• Language: English
• Subjects: Acetyl Coenzyme A - genetics; Acetyl Coenzyme A - metabolism; Acetyl-CoA carboxylase; Acetyltransferases - biosynthesis; Acetyltransferases - genetics; Acyltransferases - biosynthesis; Acyltransferases - genetics; Aspergillus nidulans; Aspergillus nidulans - enzymology; Aspergillus nidulans - genetics; Heterologous expression; Ligases - biosynthesis; Ligases - genetics; Macrolides - metabolism; Malonyl Coenzyme A - genetics; Malonyl Coenzyme A - metabolism; Malonyl-CoA; Multienzyme Complexes - biosynthesis; Multienzyme Complexes - genetics; Oxidoreductases - biosynthesis; Oxidoreductases - genetics; Penicillium - enzymology; Penicillium - genetics; Penicillium patulum; Peptide Elongation Factor 1 - genetics; Peptide Elongation Factor 1 - metabolism; Polyketides; Precursor supply; Saccharomyces cerevisiae; Saccharomyces cerevisiae - enzymology; Saccharomyces cerevisiae - genetics; Saccharomyces cerevisiae - growth & development; Saccharomyces cerevisiae Proteins - genetics; Saccharomyces cerevisiae Proteins - metabolism; Salicylates - metabolism
• ISSN: 1096-7176; 1096-7184
• DOI: 10.1016/j.ymben.2008.04.005

The heterologous production of fungal polyketides was investigated using 6-methylsalicylic acid synthase (6-MSAS) as a model polyketide synthase and Saccharomyces cerevisiae as a host. In order to improve the production of 6-MSA by enhancing the supply of precursors, the promoter of the gene ( ACC1) encoding acetyl-CoA carboxylase, which catalyzes the conversion of acetyl-CoA to malonyl-CoA, was replaced with a strong, constitutive promoter ( TEF1p) in a strain harboring two plasmids carrying the genes encoding 6-MSAS from Penicillium patulum and PPTase from Aspergillus nidulans, respectively. The strain was characterized in batch cultivations with a glucose minimal media (20 g/L), and a 60% increase in 6-MSA titer was observed compared to a strain having the native promoter in front of ACC1. The production of 6-MSA was scaled up by the cultivation in minimal media containing 50 g/L of glucose, and hereby a final titer of 554±26 mg/L of 6-MSA was obtained.