1887

Abstract

A new strain of pink facultative methylotroph, bacterium AT2, capable of growth on methylamine, trimethylamine, methanol, formate and a range of non-C substrates has been isolated. On the basis of the enzyme activities in cell-free extracts, the organism appears to have the same pathway for trimethylamine oxidation as , i.e. via trimethylamine -oxide and -methylglutamate. -Methylglutamate dehydrogenase in this organism was a ‘soluble’ enzyme (i.e. was not sedimented at 100000 in 60 min) and reacted with the electron acceptors phenazine methosulphate, 2,6-dichlorophenolindophenol, Wurster’s blue and the radical cation of 2,2′-azinodi-[3-ethylbenzthiazoline 6-sulphonate]. It was not active with NAD, ferricyanide or cytochrome The enzyme was purified to apparent homogeneity (as assessed by polyacrylamide gel electrophoresis) and was shown to contain flavin and cytochrome , both of which could be reduced by -methylglutamate. The flavin prosthetic group could be liberated by boiling and was probably FAD. The true of the enzyme for -methylglutamate was 0·33 m and for 2,6-dichlorophenolindophenol, 0·20 m.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-117-2-293
1980-04-01
2021-10-26
Loading full text...

Full text loading...

/deliver/fulltext/micro/117/2/mic-117-2-293.html?itemId=/content/journal/micro/10.1099/00221287-117-2-293&mimeType=html&fmt=ahah

References

  1. Andrews P. 1970; Estimation of molecular size and molecular weights of biological compounds by gel filtration.. Methods of Biochemical Analysis 18:1–53
    [Google Scholar]
  2. Appleby C. A., Morton R. K. 1969; Lactic dehydrogenase and cytochrome b2of baker’s yeast. Enzymic and chemical properties of the crystalline enzyme.. Biochemical Journal 73:539–550
    [Google Scholar]
  3. Bamforth C. W., Large P. J. 1975; Solubilization and partial purification of N-methylglutamate dehydrogenase from Pseudomonas aminovorans.. Biochemical Society Transactions 3:1066–1069
    [Google Scholar]
  4. Bamforth C. W., Large P. J. 1977a; Solubilization, partial purification and properties of N- methylglutamate dehydrogenase from Pseudomonas aminovorans.. Biochemical Journal 161:357–370
    [Google Scholar]
  5. Bamforth C. W., Large P. J. 1977b; The molecular size of N-methylglutamate dehydrogenase of Pseudomonas aminovorans.. Biochemical Journal 167:509–512
    [Google Scholar]
  6. Bamforth C. W., Quayle J. R. 1978; Aerobic and anaerobic growth of Paracoccus denitrificans on methanol.. Archives of Microbiology 119:91–97
    [Google Scholar]
  7. Biochemica Information 1973, 1975; 1 2 Mannheim: The Boehringer Corporation;
  8. Boulton C. A., Large P. J. 1977; Synthesis of certain assimilatory and dissimilatory enzymes during bacterial adaptation to growth on tri- methylamine.. Journal of General Microbiology 101:151–156
    [Google Scholar]
  9. Boulton C. A., Large P. J. 1979; Inactivation of trimethylamine N-oxide aldolase (demethylase) during preparation of bacterial extracts.. FEMS Microbiology Letters 5:159–162
    [Google Scholar]
  10. Bradford M. M. 1976; A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.. Analytical Biochemistry 72:248–254
    [Google Scholar]
  11. Clarke S. 1975; The size and detergent binding of membrane proteins.. Journalof Biological Chemistry 250:5459–5469
    [Google Scholar]
  12. Cleland W. W. 1963; The kinetics of enzyme- catalysed reactions with two or more substrates or products. II. Inhibition: nomenclature and theory.. Biochimica et biophysica acta 67:173–187
    [Google Scholar]
  13. Colby J., Zatman L. J. 1973; Trimethylamine metabolism in obligate and facultative methylo- trophs.. Biochemical Journal 132:101–112
    [Google Scholar]
  14. Cowan S. T., Steel K. J. 1965 Manual for the Identification of Medical Bacteria. Cambridge: Cambridge University Press;
    [Google Scholar]
  15. Cruikshank R. 1965 Medical Microbiology: A Guide to the Laboratory Diagnosis and Control of Infection, 11th edn.. Edinburgh: E. & S. Livingstone;
    [Google Scholar]
  16. Davis B. J. 1964; Disc electrophoresis. II. Method and application to human serum proteins.. Annals of the New York Academy of Sciences 121:404–427
    [Google Scholar]
  17. Den Dooren De Jong L. E. 1928; Über protamino- phage Bakterien.. Zentralblatt für Bakteriologie, Parasitenkunde, Infektionskrankheiten und Hygiene (Abteilung II) 71:193–232
    [Google Scholar]
  18. Duine J. A., Frank J., Westerling J. 1978; Purification and properties of methanol dehydrogenase from Hyphomicrobium X.. Biochimica et biophysica acta 524:277–287
    [Google Scholar]
  19. Durham D. R., Perry J. J. 1978a; Purification and characterization of a heme-containing amine dehydrogenase from Pseudomonas putida.. Journal of Bacteriology 134:837–843
    [Google Scholar]
  20. Durham D. R., Perry J. J. 1978b; Amine dehydrogenase of Pseudomonas putida: properties of the heme-prosthetic group.. Journal of Bacteriology 135:981–986
    [Google Scholar]
  21. Eady R. R., Large P. J. 1968; Purification and properties of an amine dehydrogenase from Pseudomonas AMI and its role in growth on methylamine.. Biochemical Journal 168:245–255
    [Google Scholar]
  22. Eady R. R., Jarman T. R., Large P. J. 1971; Microbial oxidation of amines. Partial purification of a mixed-function secondary-amine oxidase system from Pseudomonas aminovorans that contains an enzymically active cytochrome P-420- type haemoprotein.. Biochemical Journal 125:449–459
    [Google Scholar]
  23. Fazekas A. G., Kokai K. 1971; Extraction, purification and separation of tissue flavins for spectrophotometric determination.. Methods in Enzymology 18B:385–398
    [Google Scholar]
  24. Hersh L. B., Peterson J. A., Thompson A. A. 1971; An N-methylglutamate dehydrogenase from Pseudomonas MA.. Archives of Biochemistry and Biophysics 145:115–120
    [Google Scholar]
  25. Hersh L. B., Stark M. L., Worthen S., Fiero M. K. 1972; N-Methylglutamate dehydrogenase: kinetic studies on the solubilized enzyme.. Archives of Biochemistry and Biophysics 150:219–226
    [Google Scholar]
  26. Holloway P. W. 1972; A simple procedure for removal of Triton X-100 from protein samples.. Analytical Biochemistry 53:304–308
    [Google Scholar]
  27. Homcy C., Wrenn S., Haber E. 1978; Affinity purification of cardiac adenylate cyclase: dependence on prior hydrophobic resolution.. Proceedings of the National Academy of Sciences of the United States of America 75:59–63
    [Google Scholar]
  28. Jarman T. R. 1973 The enzymic oxidation of primary and secondary amines by Pseudomonas aminovorans. Ph.D. thesis University of Hull:
    [Google Scholar]
  29. Jarman T. R., Large P. J. 1972; Distribution of the enzymes oxidizing secondary and tertiary amines in Pseudomonas aminovorans grown on various substrates.. Journal of General Microbiology 73:205–208
    [Google Scholar]
  30. Johnson P. A., Quayle J. R. 1964; Microbial growth on Ci compounds. 6. Oxidation of methanol, formaldehyde and formate by methanol-grown Pseudomonas AMI.. Biochemical Journal 93:281–290
    [Google Scholar]
  31. Kung H.-F. 1969 Utilization of one carbon units: metabolism of methylamine by Pseudomonas MS. Ph.D. thesis Vanderbilt University, U.S.A.:
    [Google Scholar]
  32. Loginova N. V., Trotsenko Y. A. 1974; Enzymes of the methanol and methylamine metabolism in Pseudomonas methylica.. Microbiology English translation of Mikrobiologiva) 43:831–836
    [Google Scholar]
  33. Loginova N. V., Trotsenko Y. A. 1977; The metabolic pathways of methylated amines in bacteria.. In Microbial Growth on C1-Compounds Abstracts of 2nd International Symposium pp. 37–39 Skryabin G.K, Ivanov M.V, Kondratjeva E.N, Zavarzin G.V, Trotsenko Y.A, Nesterov A.I. Edited by Pushchino: Scientific Centre for Biological Research;
    [Google Scholar]
  34. Loginova N. V., Shishkina V. N., Trotsenko Y. A. 1976; Primary metabolic pathways of methylated amines in Hyphomicrobium vulgare.. Microbiology English translation of Mikrobiologiya 45:34–40
    [Google Scholar]
  35. Loginova N. V., Shishkina V. N., Filippova T. M., Trotsenko Y. A. 1977; Comparative study of the metabolism of methanol and methylamine in Hyphomicrobium vulgare 3.. Izvestiya Akademii Nauk SSSR, Seriya Biologischeskava English translation 2:235–243
    [Google Scholar]
  36. Meiberg J. B. M., Harder W. 1978; Aerobic and anaerobic metabolism of trimethylamine, di- methylamine and methylamine in Hyphomicrobium X.. Journal of General Microbiology 106:265–276
    [Google Scholar]
  37. Netrusov A. I. 1975; NAD-dependent N-methyl- glutamate dehydrogenase: a new enzyme of methylamine metabolism in methylotrophs. . Microbiology English translation of Mikro-biologiya 44:487–488
    [Google Scholar]
  38. Netrusov A. I., Rodionov Y. V., Kondratieva E. N. 1977; ATP-generation coupled with Q- compound oxidation by methylotrophic bacterium Pseudomonas sp. 2.. FEBS Letters 76:56–58
    [Google Scholar]
  39. Newby A. C., Chrambach A. 1979; Disaggregation of adenylate cyclase during polyacrylamide-gel electrophoresis in mixtures of ionic and non-ionic detergents.. Biochemical Journal 177:623–630
    [Google Scholar]
  40. O’Keeffe D. T., Anthony C. 1978; The microbial metabolism of C1 compounds. The stoicheiometry of respiration-driven proton translocation in Pseudomonas AMI and in a mutant lacking cytochrome c.. Biochemical Journal 170:561–567
    [Google Scholar]
  41. Plowman K. M. 1972 Enzyme Kinetics p. 50 New York: McGraw-Hill;
    [Google Scholar]
  42. Probst I., Wolf G., Schlegel H. G. 1979; An oxygen-binding flavohaemoprotein from Alcali- genes eutrophus. . Biochimica et biophysica acta 576:471–478
    [Google Scholar]
  43. Shaw W. V., Tsai L., Stadtman E. R. 1966; The enzymatic synthesis of N-methylglutamic acid.. Journal of Biological Chemistry 241:935–945
    [Google Scholar]
  44. Singer S. J. 1974; The molecular organization of membranes.. Annual Review of Biochemistry 43:805–833
    [Google Scholar]
  45. Strittmatter P. 1961; The nature of the flavin binding in microsomal cytochrome b5reductase.. Journal of Biological Chemistry 236:2329–2335
    [Google Scholar]
  46. Tabor C. W., Kellogg P. D. 1970; Identification of flavin adenine dinucleotide and heme in a homogeneous spermidine dehydrogenase from Serratia marcescens. . Journal of Biological Chemistry 245:5424–5433
    [Google Scholar]
  47. Thomas P. E., Lu A. Y. H., Ryan D., West S. B., Kawelek J., Levin W. 1976; Multiple forms of rat liver cytochrome P-450. Immunochemical evidence with antibody against cytochrome P-448.. Journal of Biological Chemistry 251:1385–1391
    [Google Scholar]
  48. Trotsenko Y. A., Loginova N. V. 1973; The methylamine metabolism of Pseudomonas sp.. Microbiology English translation of Mikrobiologiya 42:695–700
    [Google Scholar]
  49. Trotsenko Y. A., Loginova N. V. 1978; Pathways of metabolism of methylated amines in bacteria.. Uspekhi Mikrobiologii (Advances in Microbiology) 14:28–55 In Russian
    [Google Scholar]
  50. Van Verseveld H. W., Stouthamer A. H. 1978; Electron-transport chain and coupled oxidative phosphorylation in methanol-grown Paracoccus denitrificans.. Archives of Microbiology 118:13–20
    [Google Scholar]
  51. Vishniac W., Santer M. 1957; The thiobacilli.. Bacteriological Reviews 21:195–213
    [Google Scholar]
  52. Wallenfels K. 1962; β-Galactosidase (crystalline).. Methods in Enzymology 5:212–219
    [Google Scholar]
  53. Wang C.-S., Smith R. L. 1975; Lowry determination of protein in the presence of Triton X-100.. Analytical Biochemistry 63:414–417
    [Google Scholar]
  54. Zahler W. L. 1974; Analytical polyacrylamide gel electrophoresis and molecular weight determination.. Methods in Enzymology 32:70–81
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-117-2-293
Loading
/content/journal/micro/10.1099/00221287-117-2-293
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error