Isobutanol production in engineered Saccharomyces cerevisiae by overexpression of 2-ketoisovalerate decarboxylase and valine biosynthetic enzymes

• Published in: Bioprocess and biosystems engineering Vol. 35; no. 9; pp. 1467 - 1475
• Main Authors: Lee, Won-Heong, Seo, Seung-Oh, Bae, Yi-Hyun, Nan, Hong, Jin, Yong-Su, Seo, Jin-Ho
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Springer-Verlag 01.11.2012; Springer; Springer Nature B.V
• Subjects: 2-Acetolactate Mutase - biosynthesis; 2-Acetolactate Mutase - genetics; Acetolactate Synthase - biosynthesis; Acetolactate Synthase - genetics; Bacterial Proteins - biosynthesis; Bacterial Proteins - genetics; Biological and medical sciences; Biosynthesis; Biotechnology; Butanols - metabolism; Carboxy-Lyases - biosynthesis; Carboxy-Lyases - genetics; Chemistry; Chemistry and Materials Science; Environmental Engineering/Biotechnology; Ethanol; Food Science; Fundamental and applied biological sciences. Psychology; Gene expression; Genetic engineering; Hydro-Lyases - biosynthesis; Hydro-Lyases - genetics; Industrial and Production Engineering; Industrial Chemistry/Chemical Engineering; Lactococcus lactis - enzymology; Lactococcus lactis - genetics; Metabolic Engineering; Original Paper; Saccharomyces cerevisiae; Saccharomyces cerevisiae - enzymology; Saccharomyces cerevisiae - genetics; Valine - biosynthesis; Valine - genetics; Engineered; Valine biosynthetic enzymes; 2-Ketoisovalerate decarboxylase; Isobutanol; Ascomycota; Decarboxylase; Yeast; Enzyme; Gene overexpression; Lyases; Biosynthesis; Engineered Saccharomyces; Fungi; Carbon-carbon lyases; Valine; Carboxy-lyases; Production; cerevisiae; Saccharomyces cerevisiae
• ISSN: 1615-7591; 1615-7605
• DOI: 10.1007/s00449-012-0736-y

Engineering of Saccharomyces cerevisiae to produce advanced biofuels such as isobutanol has received much attention because this yeast has a natural capacity to produce higher alcohols. In this study, construction of isobutanol production systems was attempted by overexpression of effective 2-keto acid decarboxylase (KDC) and combinatorial overexpression of valine biosynthetic enzymes in S. cerevisiae D452-2. Among the six putative KDC enzymes from various microorganisms, 2-ketoisovalerate decarboxylase (Kivd) from L . lactis subsp. lactis KACC 13877 was identified as the most suitable KDC for isobutanol production in the yeast. Isobutanol production by the engineered S . cerevisiae was assessed in micro-aerobic batch fermentations using glucose as a sole carbon source. 93 mg/L isobutanol was produced in the Kivd overexpressing strain, which corresponds to a fourfold improvement as compared with the control strain. Isobutanol production was further enhanced to 151 mg/L by additional overexpression of acetolactate synthase (Ilv2p), acetohydroxyacid reductoisomerase (Ilv5p), and dihydroxyacid dehydratase (Ilv3p) in the cytosol.