Identification of an NADH-dependent 5-hydroxymethylfurfural-reducing alcohol dehydrogenase in Saccharomyces cerevisiae.
We report on the identification and characterization of a mutated alcohol dehydrogenase 1 from the industrial Saccharomyces cerevisiae strain TMB3000 that mediates the NADH-dependent reduction of 5-hydroxymethylfurfural (HMF) to 2,5-bis-hydroxymethylfuran. The co-factor preference distinguished this alcohol dehydrogenase from the previously reported NADPH-dependent S. cerevisiae HMF alcohol dehydrogenase Adh6. The amino acid sequence revealed three novel mutations (S109P, L116S and Y294C) that were all predicted at the vicinity of the substrate binding site, which could explain the unusual substrate specificity. Increased biomass production and HMF conversion rate were achieved in a CEN.PK S. cerevisiae strain overexpressing the mutated ADH1 gene.
Laadan
Boaz
B
Department of Applied Microbiology, Lund University, PO Box 124, SE-22100 Lund, Sweden.
Almeida
João R M
JR
Rådström
Peter
P
Hahn-Hägerdal
Bärbel
B
Gorwa-Grauslund
Marie
M
D000426:Alcohol Dehydrogenase / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D003067:Coenzymes / Q000378:metabolism; D005662:Furaldehyde / Q000031:analogs & derivatives* / Q000378:metabolism; D013058:Mass Spectrometry; D009243:NAD / Q000378:metabolism*; D010084:Oxidation-Reduction; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D013379:Substrate Specificity
D003067:Coenzymes; D029701:Saccharomyces cerevisiae Proteins; D009243:NAD; C008046:5-hydroxymethylfurfural; D005662:Furaldehyde; C516801:ADH1 protein, S cerevisiae; D000426:Alcohol Dehydrogenase