Abstract
We have purified ADHIV, a novel alcohol dehydrogenase (ADH) isozyme in the yeast Saccharomyces cerevisiae, after increasing the normally low amount of ADHIV protein in laboratory strains. This was done by overexpression of the structural gene (ADH4) on a 2micro-based multicopy vector. Characterization of the purified enzyme revealed a dimeric structure as well as a different substrate specificity and pH profile as compared to other alcohol dehydrogenase isozymes. On the other hand, we could demonstrate that ADHIV is activated by zinc ions, like the other yeast alcohol dehydrogenase isozymes, and not by ferrous ions, like a structurally similar alcohol dehydrogenase from the bacterium Zymomonas mobilis.
Publication types
-
Research Support, Non-U.S. Gov't
MeSH terms
-
Alcohol Dehydrogenase / biosynthesis
-
Alcohol Dehydrogenase / genetics*
-
Alcohol Dehydrogenase / isolation & purification
-
Enzyme Activation / drug effects
-
Fungal Proteins / biosynthesis
-
Fungal Proteins / genetics*
-
Fungal Proteins / isolation & purification
-
Hydrogen-Ion Concentration
-
Isoenzymes / biosynthesis
-
Isoenzymes / genetics*
-
Isoenzymes / isolation & purification
-
Kinetics
-
Recombinant Proteins / biosynthesis
-
Recombinant Proteins / isolation & purification
-
Saccharomyces cerevisiae / enzymology
-
Saccharomyces cerevisiae / genetics*
-
Substrate Specificity
-
Zinc / pharmacology
Substances
-
Fungal Proteins
-
Isoenzymes
-
Recombinant Proteins
-
Alcohol Dehydrogenase
-
Zinc