An investigation of the metabolism of valine to isobutyl alcohol in Saccharomyces cerevisiae.
The metabolism of valine to isobutyl alcohol in yeast was examined by 13C nuclear magnetic resonance spectroscopy and combined gas chromatography-mass spectrometry. The product of valine transamination, alpha-ketoisovalerate, had four potential routes to isobutyl alcohol. The first, via branched-chain alpha-ketoacid dehydrogenase to isobutyryl-CoA is not required for the synthesis of isobutyl alcohol because abolition of branched-chain alpha-ketoacid dehydrogenase activity in an lpd1 disruption mutant did not prevent the formation of isobutyl alcohol. The second route, via pyruvate decarboxylase, is the one that is used because elimination of pyruvate decarboxylase activity in a pdc1 pdc5 pdc6 triple mutant virtually abolished isobutyl alcohol production. A third potential route involved alpha-ketoisovalerate reductase, but this had no role in the formation of isobutyl alcohol from alpha-hydroxyisovalerate because cell homogenates could not convert alpha-hydroxyisovalerate to isobutyl alcohol. The final possibility, use of the pyruvate decarboxylase-like enzyme encoded by YDL080c, seemed to be irrelevant, because a strain with a disruption in this gene produced wild-type levels of isobutyl alcohol. Thus there are major differences in the catabolism of leucine and valine to their respective "fusel" alcohols. Whereas in the catabolism of leucine to isoamyl alcohol the major route is via the decarboxylase encoded by YDL080c, any single isozyme of pyruvate decarboxylase is sufficient for the formation of isobutyl alcohol from valine. Finally, analysis of the 13C-labeled products revealed that the pathways of valine catabolism and leucine biosynthesis share a common pool of alpha-ketoisovalerate.
Dickinson
J R
JR
School of Pure & Applied Biology, University of Wales, Cardiff, CF1 3TL, United Kingdom.
Harrison
S J
SJ
Hewlins
M J
MJ
D000440:Butanols / Q000378:metabolism*; D002247:Carbon Isotopes; D002455:Cell Division / Q000235:genetics; D005656:Fungal Proteins / Q000378:metabolism; D008401:Gas Chromatography-Mass Spectrometry; D045782:Hemiterpenes; D007651:Keto Acids / Q000378:metabolism; D007930:Leucine / Q000096:biosynthesis; D009682:Magnetic Resonance Spectroscopy; D011767:Pyruvate Decarboxylase / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D014633:Valine / Q000378:metabolism*
D000440:Butanols; D002247:Carbon Isotopes; D005656:Fungal Proteins; D045782:Hemiterpenes; D007651:Keto Acids; C040507:isobutyl alcohol; C001505:alpha-ketoisovalerate; D011767:Pyruvate Decarboxylase; D007930:Leucine; D014633:Valine