Characterization of Ypr1p from Saccharomyces cerevisiae as a 2-methylbutyraldehyde reductase.
The metabolism of aldehydes and ketones in yeast is important for biosynthetic, catabolic and detoxication processes. Aldo-keto reductases are a family of enzymes that are able to reduce aldehydes and ketones. The roles of individual aldo-keto reductases in yeast has been difficult to determine because of overlapping substrate specificities of these enzymes. In this study, we have cloned, expressed and characterized the aldo-keto reductase Ypr1p from the yeast Saccharomyces cerevisiae and we describe its substrate specificity. The enzyme displays high specific activity towards 2-methylbutyraldehyde, as well as other aldehydes such as hexanal. It exhibits extremely low activity as a glycerol dehydrogenase. The enzyme functions over a wide pH range and uses NADPH as co-factor. In comparison to other mammalian and yeast aldo-keto reductases, Ypr1p has relatively high affinity for D,L-glyceraldehyde (1.08 mM) and hexanal (0.39 mM), but relatively low affinity for 4-nitrobenzaldehyde (1.07 mM). It displays higher specific activity for 2-methylbutyraldehyde than does yeast alcohol dehydrogenase and has a K(m) for 2-methyl butyraldehyde of 1.09 mM. The enzyme is expressed during growth on glucose, but its levels are rapidly induced by osmotic and oxidative stress. Yeast in which the YPR1 gene has been deleted possess 50% lower 2-methylbutyraldehyde reductase activity than the wild-type strain. This suggests that the enzyme may contribute to 2-methyl butyraldehyde reduction in vivo. It may therefore play a role in isoleucine catabolism and fusel alcohol formation and may influence flavour formation by strains of brewing yeast.
Ford
Gordon
G
Department of Pharmaceutical Sciences, University of Strathclyde, 204 George Street, Glasgow G1 1XW, UK.
Ellis
Elizabeth M
EM
D000429:Alcohol Oxidoreductases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000449:Aldehyde Reductase; D000447:Aldehydes / Q000737:chemistry / Q000378:metabolism*; D000074408:Aldo-Keto Reductases; D017353:Gene Deletion; D007700:Kinetics; D009997:Osmotic Pressure; D018384:Oxidative Stress; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D013379:Substrate Specificity; D013401:Sugar Alcohol Dehydrogenases / Q000378:metabolism
D000447:Aldehydes; D029701:Saccharomyces cerevisiae Proteins; D000429:Alcohol Oxidoreductases; D013401:Sugar Alcohol Dehydrogenases; D000074408:Aldo-Keto Reductases; C466911:YPR1protein, S cerevisiae; D000449:Aldehyde Reductase; C020672:glycerol dehydrogenase; C018475:butyraldehyde