Abstract
The α-isopropylmalate synthase of the chemolithoautotrophic Alcaligenes eutrophus H16 is apparently a soluble enzyme but is strongly adsorbed to cell particles in ruptured cell suspensions. This was not observed with α-acetohydroxy acid synthase or threonine deaminase. The formation of these regulatory enzymes of the branched chain amino acid biosynthesis pathway generally decreased with decreased growth rates. The addition of 5 mM valine plus isoleucine with and without 5 mM threonine caused a 6.6- and a 4-fold increase, respectively, in the formation of active α-isopropylmalate synthase, but caused a strong decrease in the α-actohydroxy acid synthase. The level of active α-isopropylmalate synthase is apparently regulated by the level of leucine; whereas, the level of the α-acetohydroxy acid synthase and threonine deaminase is influenced by the presence of several amino acids. A catabolic threonine deaminase was not encountered.
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Abbreviations
- IRS:
-
α-Isopropylamalate
- AHA:
-
α-acetohydroxy acid
- TDA:
-
throninedeaminase
References
Allison MJ, Baetz AL, Wiegel J (1984) Alternative pathways for biosynthesis of leucine and other amino acids in Bacteroides ruminicola and Bacteroides fragilis. Appl Environ Microbiol 48:1111–1117
Beisenherz G, Balze HJ, Bücher T, Czok R, Garbade HK, Meyer-Ahrendt E, Pfleiderer G (1953) Diphosphofructose-Aldolase, Phosphoglyceraldehyd-Dehydrogenase, Milchsäure-Dehydrogenase, Glycerophosphat-Dehydrogenase and Pyruvat-Kinase aus Kaninchenmuskulatur in einem Arbeitsgang. Z Naturf 8B:555–557
Caivo JM (1983) Leucine biosynthesis in prokaryotes. In: Herrman KM, Somerville RL (eds) Amino acids. Biosynthesis and genetic regulation. Addison-Wesley, London, pp 267–284
Calvo JM, Bartholomew JC, Stieglitz BJ (1969) Fluorometric assay of enzymatic reactions involving acetyl coenzyme A in aldol condensations. Anal Biochem 28:164–181
Donald RA, Satyanarayana T, Kaplan JG (1974) Biosynthesis of branched-chain amino acids in Schizosaccharomyces pombe. Regulation of the enzymes involved in isoleucine, valine and leucine synthesis. Can J Biochem 52:51–59
Gollop N, Chipman DM, Bakar Z (1983) Inhibition of acetohydroxy acid synthase by leucine. Biochim Biophys Acta 748:34–39
Hill FF (1968) Regulation der Leucinsynthese and Trifluorleucinresistenz bei Hydrogenomonas H16. Ph. D. Thesis, University Göttingen
Hill FF, Schlegel HG (1969) Regulationsdefekte bei trifluorleucinresistenten Mutanten von Hydrogenomonas H16. Arch Mikrobiol 68:18–31
Holzer H, Boll M, Cennamo C (1963) Zur Biochemie der Threonine-Desaminase aus Hefe. Angew Chem 75:894–900
Hsu Y-P, Kohlhaw GB, Niederberger P (1982) Evidence that α-isopropyl-malate synthase of Saccharomyces cerevisiae is under the “general” control of amino acid biosynthesis. J Bacteriol 150:969–972
Kohlhaw GB (1983) Regulation of leucine biosynthesis in lower eukaryotes. In: Herrmann KM, Sommerville RL (eds) Amino acids. Biosynthesis and genetic regulation. Addison-Wesley, London, pp 285–300
Miflin BJ, Cave PR (1972) The control of leucine, isoleucine and valine biosynthesis in a range of higher plants. J Exp Bot 23:511–516
Niederberger P, Hütter R (1980) The role of the general control of amino acid biosynthesis of Saccharomyces cerevisiae in vivo. Experientia 36:1456
Probst I (1975) Untersuchungen über das elektronentransport-System von Alcaligenes eutrophus Stamm H16. Ph. D. Thesis, University Göttingen
Reh M (1967) Regulation der Synthese von Isoleucin und Valin durch Hydrogenomonas H16. Ph. D. Thesis, University Göttingen
Reh M, Schlegel HG (1969) Die Biosynthese von Isoleucin and Valin in Hydrogenomonas H16. Arch Mikrobiol 67:110–127
Rennert OM, Anker SH (1963) Incorporation of 5′,5′,5′-trifluoroleucine into proteins of E. coli. Biochemistry 2:471–476
Ryan ED, Kohlhaw GB (1974) Subcellular localization of isoleucine-valine biosynthetic enzymes in yeast. J Bacteriol 120:631–637
Ryan ED, Tracy JW, Kohlhaw GB (1973) Subcellular localization of the leucine biosynthetic enzymes in yeast. J Bacteriol 116:222–225
Singer RA, Doolittle WF (1975) Leucine biosynthesis in the bluegreen bacterium Anacystis nidulans. J Bacteriol 124:810–814
Stieglitz BI, Calvo JM (1974) Distribution of the isopropylmalate pathway to leucine among diverse bacteria. J Bacteriol 118:935–941
Tracy JW, Kohlhaw GB (1975) Reversible, coenzyme A-mediated inactivation of biosynthetic condensing enzymes in yeast: A possible regulatory mechanism. Proc Natl Acad Sci USA 72:1802–1806
Umbarger HE (1983) The biosynthesis of isoleucine and valine and its regulation. In: Herrman KM, Somerville RL (eds) Amino acids. Biosynthesis and genetic regulation. Addison-Wesley, London, pp 245–266
Umbarger EU, Brown B (1958) Isoleucine and valine metabolism in Escherichia coli. VIII. The formation of acetolactate. J Biol Chem 233:1156–1160
Wiegel J (1973) Die α-Isopropylmalat-Synthase in Hydrogenomonas eutropha H16. Ph. D. Thesis, University Göttingen
Wiegel J (1978) Mn++-specific reactivation of EDTA inactivated α-isopropylmalate synthase from Alcaligenes eutrophus H16. Biochem Biophys Res Commun 82:907–912
Wiegel J (1981) α-Isopropylmalate synthase as a marker for the leucine biosynthetic pathway in several Clostridia and in Bacteroides fragilis. Arch Microbiol 130:385–390
Wiegel J, Schlegel HG (1977a) α-Isopropylmalate synthase from Alcaligenes eutrophus H16. I. Purification and general properties. Arch Microbiol 112:239–246
Wiegel J, Schlegel HG (1977b) α-Isopropylmalate synthase from Alcaligenes eutrophus H16. II. Substrate specificity and kinetics. Arch Microbiol 112:247–254
Wiegel J, Schlegel HG (1977c) α-Isopropylmalatesynthase in Alcaligenes eutrophus H16. III. End product inhibition and its relief by valine and isoleucine. Arch Microbiol 114:203–210
Wiegel J, Schlegel HG (1977d) Leucine biosynthesis.: Effect of branched-chain amino acids and threonine on α-isopropylmalate synthase activity from aerobic and anaerobic microorganisms. Biochem Syst Ecol 5:169–176
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This paper is dedicated to Professor H. G. Schlegel, University Göttingen, on the occasion of his 60th birthday. I am grateful to a great teacher and scientist, who in his unique way stimulated enthusiasm and fascination in microbiology in his students throughout the years
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Wiegel, J. Leucine biosynthesis in Alcaligenes eutrophus H16: Influence of amino acid additions on the formation of active α-isopropylmalate synthase and α-acetohydroxy acid synthase. Arch. Microbiol. 142, 194–199 (1985). https://doi.org/10.1007/BF00447067
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DOI: https://doi.org/10.1007/BF00447067