Engineering yeast alcohol dehydrogenase. Replacing Trp54 by Leu broadens substrate specificity.
Analysis of a crystal structure of alcohol dehydrogenase (Adh) from horse liver suggests that Trp54 in the homologous yeast alcohol dehydrogenase prevents the yeast enzyme from efficiently catalysing the oxidation of long-chain primary alcohols with branching at the 4 position (e.g. 4-methyl-1-pentanol, cinnamyl alcohol). This residue has been altered to Leu by site-directed mutagenesis. The alteration yields an enzyme that serves as an effective catalyst for both longer straight-chain primary alcohols and branched chain alcohols.
Weinhold
E G
EG
Department of Chemistry, ETH, Zurich, Switzerland.
Benner
S A
SA
D000426:Alcohol Dehydrogenase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000438:Alcohols / Q000378:metabolism; D001665:Binding Sites; D007700:Kinetics; D007930:Leucine / Q000737:chemistry; D016297:Mutagenesis, Site-Directed; D015202:Protein Engineering; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics; D013379:Substrate Specificity; D014364:Tryptophan / Q000737:chemistry
D000438:Alcohols; D014364:Tryptophan; D000426:Alcohol Dehydrogenase; D007930:Leucine