Abstract
Cell wall-membrane preparations of Escherichia coli, prepared by the ethylenediaminetetraacetic acid-lysozyme method, contain enzymes which catalyze the oxidation of d-alanine and, to a lesser extent, l-alanine into pyruvate and ammonia without the formation of hydrogen peroxide. The kinetic parameters were (i) pH optima of 8.3 to 8.4 for l- and d-alanine and (ii) a Km value of 6.6 ± 0.2 mM for d-alanine. Several coenzymes were without effect when added to the reaction mixture. The participation of d-alanine oxidase in the oxidation of l-alanine was demonstrated. The evidence is based on (i) results of cellular fractionation; (ii) labeling experiments; (iii) inhibition studies with aminooxyacetate and cycloserine; (iv) denaturation experiments; and (v) demonstration of the presence of an active racemase.
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Selected References
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- BARBIERI P., DI MARCO A., FUOCO L., RUSCONI A. Investigations on the mode of action of cycloserine upon protein synthesis in Escherichia coli. Biochem Pharmacol. 1960 May;3:101–109. doi: 10.1016/0006-2952(60)90026-5. [DOI] [PubMed] [Google Scholar]
- De Duve C., Baudhuin P. Peroxisomes (microbodies and related particles). Physiol Rev. 1966 Apr;46(2):323–357. doi: 10.1152/physrev.1966.46.2.323. [DOI] [PubMed] [Google Scholar]
- GREENSTEIN J. P., BIRNBAUM S. M., OTEY M. C. Optical and enzymatic characterization of amino acids. J Biol Chem. 1953 Sep;204(1):307–321. [PubMed] [Google Scholar]
- Gordon A. S., Lombardi F. J., Kaback H. R. Solubilization and partial purification of amino acid-specific components of the D-lactate dehydrogenase-coupled amino acid-transport systems (E. coli-cell membranes-sephadex-detergent-solubilized-vesicles). Proc Natl Acad Sci U S A. 1972 Feb;69(2):358–362. doi: 10.1073/pnas.69.2.358. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gryder R. M., Adams E. Inducible degradation of hydroxyproline in Pseudomonas putida: pathway regulation and hydroxyproline uptake. J Bacteriol. 1969 Jan;97(1):292–306. doi: 10.1128/jb.97.1.292-306.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
- JAKOBY W. B. Aldehyde oxidation. I. Dehydrogenase from Pseudomonas fluorescens. J Biol Chem. 1958 May;232(1):75–87. [PubMed] [Google Scholar]
- KEILIN D., HARTREE E. F. Catalase, peroxidase and metmyoglobin as catalysts of coupled peroxidatic reactions. Biochem J. 1955 Jun;60(2):310–325. doi: 10.1042/bj0600310. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lambert M. P., Neuhaus F. C. Factors affecting the level of alanine racemase in Escherichia coli. J Bacteriol. 1972 Mar;109(3):1156–1161. doi: 10.1128/jb.109.3.1156-1161.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
- MCGILVERY R. W., MOKRASCH L. C. Purification and properties of fructose-1, 6-diphosphatase. J Biol Chem. 1956 Aug;221(2):909–917. [PubMed] [Google Scholar]
- Martinez-Carrion M., Jenkins W. T. D-Alanine-D-glutamate transaminase. I. Purification and characterization. J Biol Chem. 1965 Sep;240(9):3538–3546. [PubMed] [Google Scholar]
- NORTON J. E., BULMER G. S., SOKATCH J. R. THE OXIDATION OF D-ALANINE BY CELL MEMBRANES OF PSEUDOMONAS AERUGINOSA. Biochim Biophys Acta. 1963 Oct 8;78:136–147. doi: 10.1016/0006-3002(63)91619-6. [DOI] [PubMed] [Google Scholar]
- Neims A. H., Hellerman L. Flavoenzyme catalysis. Annu Rev Biochem. 1970;39:867–888. doi: 10.1146/annurev.bi.39.070170.004251. [DOI] [PubMed] [Google Scholar]
- Raunio R. P., Jenkins W. T. D-alanine oxidase form Escherichia coli: localization and induction by L-alanine. J Bacteriol. 1973 Aug;115(2):560–566. doi: 10.1128/jb.115.2.560-566.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rosso G., Takashima K., Adams E. Coenzyme content of purified alanine racemase from Pseudomonas. Biochem Biophys Res Commun. 1969 Jan 6;34(1):134–140. doi: 10.1016/0006-291x(69)90539-7. [DOI] [PubMed] [Google Scholar]
- SAITO M. [Assimilatory mechanism of D-amino acid of Mycobacterium avium. I. Oxidation of amino acid]. Kekkaku. 1956 May;31(5):267–271. [PubMed] [Google Scholar]
- Soda K. Microdetermination of D-amino acids and D-amino acid oxidase activity with 3,methyl-2-benzothiazolone hydrazone hydrochloride. Anal Biochem. 1968 Oct 24;25(1):228–235. doi: 10.1016/0003-2697(68)90095-x. [DOI] [PubMed] [Google Scholar]
- VYSHEPAN E. D., IVANOVA K. I., LEDNEVA R. K. [Formation and deamination of alanine in E. coli]. Biokhimiia. 1961 Jul-Aug;26:758–765. [PubMed] [Google Scholar]
- WOOD W. A., GUNSALUS I. C. D-Alanine formation; a racemase in Streptococcus faecalis. J Biol Chem. 1951 May;190(1):403–416. [PubMed] [Google Scholar]