Volume 272, Number 43, Issue of October 24, 1997 pp. 26871-26878
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.

A ¹³C Nuclear Magnetic Resonance Investigation of the Metabolism of Leucine to Isoamyl Alcohol in Saccharomyces cerevisiae

(Received for publication, June 17, 1997)

J. Richard Dickinson , Margaret M. Lanterman ^¶ , Dean J. Danner ^¶ , Bruce M. Pearson , Pascual Sanz ^** , Scott J. Harrison and Michael J. E. Hewlins

From the  School of Pure & Applied Biology, University of Wales, Cardiff CF1 3TL, United Kingdom, the ^¶ Department of Genetics, Emory University School of Medicine, Atlanta, Georgia 30322, the  Insitute of Food Research, Norwich NR4 7UA, United Kingdom, the ^** Consejo Superior de Investigaciones Científicas, Instituto de Agroquimica y Tecnologia de Alimentos, Valencia, Spain, and the  Department of Chemistry, University of Wales, Cardiff CF1 3TB, United Kingdom

The metabolism of leucine to isoamyl alcohol in yeast was examined by ¹³C nuclear magnetic resonance spectroscopy. The product of leucine transamination, -ketoisocaproate had four potential routes to isoamyl alcohol. The first, via branched-chain -keto acid dehydrogenase to isovaleryl-CoA with subsequent conversion to isovalerate by acyl-CoA hydrolase operates in wild-type cells where isovalerate appears to be an end product. This pathway is not required for the synthesis of isoamyl alcohol because abolition of branched-chain -keto acid dehydrogenase activity in an lpd1 disruption mutant did not prevent the formation of isoamyl alcohol. A second possible route was via pyruvate decarboxylase; however, elimination of pyruvate decarboxylase activity in a pdc1 pdc5 pdc6 triple mutant did not decrease the levels of isoamyl alcohol produced. A third route utilizes -ketoisocaproate reductase (a novel activity in Saccharomyces cerevisiae) but with no role in the formation of isoamyl alcohol from -hydroxyisocaproate because cell homogenates could not convert -hydroxyisocaproate to isoamyl alcohol. The final possibility was that a pyruvate decarboxylase-like enzyme encoded by YDL080c appears to be the major route of decarboxylation of -ketoisocaproate to isoamyl alcohol although disruption of this gene reveals that at least one other unidentified decarboxylase can substitute to a minor extent.

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