Abstract
Hydroxypyruvate reductase was purified to homogeneity from the facultative methylotroph Methylobacterium extorquens AM1. It has a molecular mass of about 71 kDa, and it consists of two identical subunits with a molecular mass of about 37 kDa. This enzyme uses both NADH (Km = 0.04 mM) and NADPH (Km = 0.06 mM) as cofactors, uses hydroxypyruvate (Km = 0.1 mM) and glyoxylate (Km = 1.5 mM) as the only substrates for the forward reaction, and carries out the reverse reaction with glycerate (Km = 2.6 mM) only. It was not possible to detect the conversion of glycolate to glyoxylate, a proposed role for this enzyme. Kinetics and inhibitory studies of the enzyme from M. extorquens AM1 suggest that hydroxypyruvate reductase is not a site for regulation of the serine cycle at the level of enzyme activity.
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Selected References
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- Anderson D. J., Lidstrom M. E. The moxFG region encodes four polypeptides in the methanol-oxidizing bacterium Methylobacterium sp. strain AM1. J Bacteriol. 1988 May;170(5):2254–2262. doi: 10.1128/jb.170.5.2254-2262.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dunstan P. M., Anthony C., Drabble W. T. Microbial metabolism of C 1 and C 2 compounds. The role of glyoxylate, glycollate and acetate in the growth of Pseudomonas AM1 on ethanol and on C 1 compounds. Biochem J. 1972 Jun;128(1):107–115. doi: 10.1042/bj1280107. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fulton G. L., Nunn D. N., Lidstrom M. E. Molecular cloning of a malyl coenzyme A lyase gene from Pseudomonas sp. strain AM1, a facultative methylotroph. J Bacteriol. 1984 Nov;160(2):718–723. doi: 10.1128/jb.160.2.718-723.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Husic D. W., Tolbert N. E. NADH:hydroxypyruvate reductase and NADPH:glyoxylate reductase in algae: partial purification and characterization from Chlamydomonas reinhardtii. Arch Biochem Biophys. 1987 Feb 1;252(2):396–408. doi: 10.1016/0003-9861(87)90046-4. [DOI] [PubMed] [Google Scholar]
- Izumi Y., Yoshida T., Kanzaki H., Toki S., Miyazaki S. S., Yamada H. Purification and characterization of hydroxypyruvate reductase from a serine-producing methylotroph, Hyphomicrobium methylovorum GM2. Eur J Biochem. 1990 Jun 20;190(2):279–284. doi: 10.1111/j.1432-1033.1990.tb15573.x. [DOI] [PubMed] [Google Scholar]
- Kalb V. F., Jr, Bernlohr R. W. A new spectrophotometric assay for protein in cell extracts. Anal Biochem. 1977 Oct;82(2):362–371. doi: 10.1016/0003-2697(77)90173-7. [DOI] [PubMed] [Google Scholar]
- Kohn L. D., Jakoby W. B. Tartaric acid metabolism. VII. Crystalline hydroxypyruvate reductase (D-glycerate dehydrogenase). J Biol Chem. 1968 May 25;243(10):2494–2499. [PubMed] [Google Scholar]
- Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
- Large P. J., Quayle J. R. Microbial growth on C(1) compounds. 5. Enzyme activities in extracts of Pseudomonas AM1. Biochem J. 1963 May;87(2):386–396. doi: 10.1042/bj0870386. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lidstrom M. E., Stirling D. I. Methylotrophs: genetics and commercial applications. Annu Rev Microbiol. 1990;44:27–58. doi: 10.1146/annurev.mi.44.100190.000331. [DOI] [PubMed] [Google Scholar]
- McNerney T., O'connor M. L. Regulation of enzymes associated with C-1 metabolism in three facultative methylotrophs. Appl Environ Microbiol. 1980 Aug;40(2):370–375. doi: 10.1128/aem.40.2.370-375.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nunn D. N., Lidstrom M. E. Isolation and complementation analysis of 10 methanol oxidation mutant classes and identification of the methanol dehydrogenase structural gene of Methylobacterium sp. strain AM1. J Bacteriol. 1986 May;166(2):581–590. doi: 10.1128/jb.166.2.581-590.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nunn D. N., Lidstrom M. E. Phenotypic characterization of 10 methanol oxidation mutant classes in Methylobacterium sp. strain AM1. J Bacteriol. 1986 May;166(2):591–597. doi: 10.1128/jb.166.2.591-597.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- PEEL D., QUAYLE J. R. Microbial growth on C1 compounds. I. Isolation and characterization of Pseudomonas AM 1. Biochem J. 1961 Dec;81:465–469. doi: 10.1042/bj0810465. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Taylor I. J., Anthony C. Acetyl-CoA production and utilization during growth of the facultative methylotroph Pseudomonas AM1 on ethanol, malonate and 3-hydroxybutyrate. J Gen Microbiol. 1976 Jul;95(1):134–143. doi: 10.1099/00221287-95-1-134. [DOI] [PubMed] [Google Scholar]
- Titus D. E., Hondred D., Becker W. M. Purification and characterization of hydroxypyruvate reductase from cucumber cotyledons. Plant Physiol. 1983 Jun;72(2):402–408. doi: 10.1104/pp.72.2.402. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tolbert N. E., Yamazaki R. K., Oeser A. Localization and properties of hydroxypyruvate and glyoxylate reductases in spinach leaf particles. J Biol Chem. 1970 Oct 10;245(19):5129–5136. [PubMed] [Google Scholar]
- Utting J. M., Kohn L. D. Structural, kinetic, and renaturation properties of an induced hydroxypyruvate reductase from Pseudomonas acidovorans. J Biol Chem. 1975 Jul 10;250(13):5233–5242. [PubMed] [Google Scholar]
- WILLIS J. E., SALLACH H. J. Evidence for a mammalian D-glyceric dehydrogenase. J Biol Chem. 1962 Mar;237:910–915. [PubMed] [Google Scholar]
- Whitaker J. R., Granum P. E. An absolute method for protein determination based on difference in absorbance at 235 and 280 nm. Anal Biochem. 1980 Nov 15;109(1):156–159. doi: 10.1016/0003-2697(80)90024-x. [DOI] [PubMed] [Google Scholar]