title: 2-Butanol and butanone production in Saccharomyces cerevisiae through combination of a B12 dependent dehydratase and a secondary alcohol dehydrogenase using a TEV-based expression system.
abstract: 2-Butanol and its chemical precursor butanone (methyl ethyl ketone--MEK) are chemicals with potential uses as biofuels and biocommodity chemicals. In order to produce 2-butanol, we have demonstrated the utility of using a TEV-protease based expression system to achieve equimolar expression of the individual subunits of the two protein complexes involved in the B12-dependent dehydratase step (from the pdu-operon of Lactobacillus reuteri), which catalyze the conversion of meso-2,3-butanediol to butanone. We have furthermore identified a NADH dependent secondary alcohol dehydrogenase (Sadh from Gordonia sp.) able to catalyze the subsequent conversion of butanone to 2-butanol. A final concentration of 4±0.2 mg/L 2-butanol and 2±0.1 mg/L of butanone was found. A key factor for the production of 2-butanol was the availability of NADH, which was achieved by growing cells lacking the GPD1 and GPD2 isogenes under anaerobic conditions.
authors: Department of Chemical and Biological Engineering, System and Synthetic Biology, Chalmers University of Technology, Gothenburg, Sweden.
authors: Payam
authors: P
authors: Ghiaci
authors: Department of Chemical and Biological Engineering, System and Synthetic Biology, Chalmers University of Technology, Gothenburg, Sweden.
authors: Joakim
authors: J
authors: Norbeck
authors: Department of Chemical and Biological Engineering, System and Synthetic Biology, Chalmers University of Technology, Gothenburg, Sweden.
authors: Christer
authors: C
authors: Larsson
mesh_terms: D000429:Alcohol Oxidoreductases / Q000235:genetics / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D053898:Biosynthetic Pathways; D015153:Blotting, Western; D000440:Butanols / Q000378:metabolism*; D002074:Butanones / Q000378:metabolism*; D010450:Endopeptidases / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D005818:Genetic Engineering / Q000379:methods; D033601:Gordonia Bacterium / Q000201:enzymology / Q000235:genetics; D007218:Industrial Microbiology / Q000379:methods; D052200:Limosilactobacillus reuteri / Q000201:enzymology / Q000235:genetics; D008969:Molecular Sequence Data; D009876:Operon / Q000235:genetics; D011408:Propanediol Dehydratase / Q000235:genetics / Q000378:metabolism*; D015203:Reproducibility of Results; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D014805:Vitamin B 12 / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D000440:Butanols; D002074:Butanones; C043958:2-butanol; D000429:Alcohol Oxidoreductases; C084155:alcohol dehydrogenase (NAD(P)+); D010450:Endopeptidases; C099488:TEV protease; D011408:Propanediol Dehydratase; D014805:Vitamin B 12