Production of the polyketide 6-MSA in yeast engineered for increased malonyl-CoA supply.
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.
Center for Microbial Biotechnology, Technical University of Denmark, Department of Systems Biology, Denmark.
Songsak
S
Wattanachaisaereekul
Anna Eliasson
AE
Lantz
Michael Lynge
ML
Nielsen
Jens
J
Nielsen
D000105:Acetyl Coenzyme A / Q000235:genetics / Q000378:metabolism; D000123:Acetyltransferases / Q000096:biosynthesis* / Q000235:genetics; D000217:Acyltransferases / Q000096:biosynthesis* / Q000235:genetics; D001233:Aspergillus nidulans / Q000201:enzymology / Q000235:genetics; D008025:Ligases / Q000096:biosynthesis* / Q000235:genetics; D018942:Macrolides / Q000378:metabolism*; D008316:Malonyl Coenzyme A / Q000235:genetics / Q000378:metabolism; D009097:Multienzyme Complexes / Q000096:biosynthesis* / Q000235:genetics; D010088:Oxidoreductases / Q000096:biosynthesis* / Q000235:genetics; D010407:Penicillium / Q000201:enzymology / Q000235:genetics; D020648:Peptide Elongation Factor 1 / Q000235:genetics / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D012459:Salicylates / Q000378:metabolism*
D018942:Macrolides; D009097:Multienzyme Complexes; D020648:Peptide Elongation Factor 1; D029701:Saccharomyces cerevisiae Proteins; D012459:Salicylates; C410826:TEF1 protein, S cerevisiae; D008316:Malonyl Coenzyme A; D000105:Acetyl Coenzyme A; D010088:Oxidoreductases; D000217:Acyltransferases; D000123:Acetyltransferases; C055210:aminoglycoside N1-acetyltransferase; C021587:6-methylsalicylic acid synthetase; D008025:Ligases; C005236:6-methylsalicylic acid