1887

Abstract

By techniques involving differential centrifugation and specific precipitation with CaCI, it was shown that dimethylamine and trimethylamine mono-oxygenase activities co-sediment with NADPH-cytochrome reductase activity in sphaeroplast lysates of grown on trimethylamine as sole nitrogen source. Since the active fraction also contained low levels of cytochromes P-450 and P-420, it was concluded that the two amine mono-oxygenases are located in the smooth endoplasmic reticulum and thus end up in the microsomal fraction on cell fractionation. Ten to twenty-fold enrichment of mono-oxygenase specific activity could be achieved by separation of activity from soluble protein by centrifugation or gel filtration. Cell-free extracts prepared in the absence of FAD showed only very low mono-oxygenase activity for either substrate. Some activity could be restored by addition of flavin nucleotides: there was a fivefold stimulation by FAD and a fourfold stimulation by FMN. All trimethylamine mono-oxygenase activity was lost when a partially purified preparation containing both activities was incubated for more than 24 h at 0°C, suggesting that separate enzymes are responsible for the oxidation of secondary and tertiary amines. The enzyme preparation oxidized a wide range of secondary alkylamines up to dibutylamine and tertiary alkylamines up to tributylamine. Primary amines, choline, di-and triethanolamine, spermine, spermidine and substituted anilines were not oxidized. NADH had a lower apparent value and higher value than NADPH. Secondary and tertiary alkylamines containing more than one kind of alkyl group gave more than one kind of aldehyde on oxidation. Stoicheiometry determinations showed a consumption of 1 mol NAD(P)H and 1 mol O per mol aldehyde formed. Carbon monoxide, cyanide, proadifen hydrochloride (SKF 525-A), mercurials and mercaploethanol all inhibited both activities.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-130-10-2577
1984-10-01
2021-08-04
Loading full text...

Full text loading...

/deliver/fulltext/micro/130/10/mic-130-10-2577.html?itemId=/content/journal/micro/10.1099/00221287-130-10-2577&mimeType=html&fmt=ahah

References

  1. Anthony C. 1982 The Biochemistry of Methylotrophs pp. 195–218 London: Academic Press;
    [Google Scholar]
  2. Barnett J.A., Payne R.W., Yarrow D. 1983 Yeasts: Characteristics and Identification pp. 317–318 Cambridge: Cambridge University Press;
    [Google Scholar]
  3. Bartels P.D., Jensen P.K. 1979; Role of AMP in regulation of the citric acid cycle in mitochondria from bakers’ yeast. Biochimica et biophysica acta 582:246–259
    [Google Scholar]
  4. Bickel M.H. 1971; N-Oxide formation and related reactions in drug metabolism. Xenobiotica 1:313–319
    [Google Scholar]
  5. Boulton C.A., Large P.J. 1979; Properties of Pseudomonas AMI primary-amine dehydrogenase immobilized on agarose. Biochimica et biophysica acta 570:22–30
    [Google Scholar]
  6. Boulton C.A., Crabbe M.J.C., Large P.J. 1974; Microbial oxidation of amines. Partial purification of a trimethylamine monooxygenase from Pseudomonas aminovorans and its role in growth on trimethylamine. Biochemical Journal 140:253–263
    [Google Scholar]
  7. Bradford M.M. 1976; A rapid and sensitive method for the quantitative determination of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72:248–254
    [Google Scholar]
  8. Brook D.F., Large P.J. 1975; Inhibition by carbon monoxide of the secondary-amine monooxygenase of Pseudomonas aminovorans and the photochemical action spectrum for its reversal. European Journal of Biochemistry 55:601–609
    [Google Scholar]
  9. Colby J., Zatman L.J. 1973; Trimethylamine metabolism in obligate and facultative methylotrophs. Biochemical Journal 132:101–112
    [Google Scholar]
  10. Delaissé J.-M., Martin P., Verheyen-Bouvy M.-F., Nyss E.J. 1981; Subcellular distribution of enzymes in the yeast Saccharomycopsis lipolytica, grown on n-hexadecane, with special reference to the ω-hydroxylase. Biochimica et biophysica acta 676:77–90
    [Google Scholar]
  11. Dubin D.T. 1960; The assay and characterization of amines by means of 2,4-dinitrofluorobenzene. Journal of Biological Chemistry 235:783–786
    [Google Scholar]
  12. Eady R.R., Jarman R.T., 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]
  13. Green J., Large P.J. 1983a; Oxidation of dimethylamine and trimethylamine in methazotrophic yeasts by microsomal mono-oxygenases sensitive to carbon monoxide. Biochemical and Biophysical Research Communications 113:900–907
    [Google Scholar]
  14. Green J., Large P.J. 1983b; Cell-free oxidation of dimethylamine by a mono-oxygenase in the methazotrophic yeast Candida utilis. Biochemical Society Transactions 11:786
    [Google Scholar]
  15. Green J., Large P.J. 1984; Regulation of the key enzymes of methylated amine metabolism in Candida boidinii. Journal of General Microbiology 130:1947–1959
    [Google Scholar]
  16. Haywood G.W., Large P.J. 1981; Microbial oxidation of amines. Distribution, purification and properties of two primary-amine oxidases from the yeast Candida boidinii grown on amines as sole nitrogen source. Biochemical Journal 199:187–201
    [Google Scholar]
  17. Hlavica P. 1982; Biological oxidation of nitrogen in organic compounds and disposition of N-oxidized products. CRC Critical Reviews of Biochemistry 12:39–101
    [Google Scholar]
  18. Holley R.A., Kidby D.K. 1973; Role of vacuoles and vesicles in extracellular enzyme secretion from yeast. Canadian Journal of Microbiology 19:113–117
    [Google Scholar]
  19. Hou C.T., Patel R.N., Barnabe N. 1982; Identification of an NADH-linked formaldehyde-reducing enzyme from methanol-grown Pichia pastoris NRRL Y-7556. FEMS Microbiology Letters 15:159–163
    [Google Scholar]
  20. Huang A.H.C., Trelease R.N., Moore T.S.JR 1983 Plant Peroxisomes pp. 87–155 New York: Academic Press;
    [Google Scholar]
  21. Jenkins R.O., Cartledge T.G., Lloyd D. 1983; Subcellular fractionation of Candida stellatoidea aftergrowth with glucose or n-hexadecane. Journal of General Microbiology 129:1171–1185
    [Google Scholar]
  22. Käppeli O., Sauer M., Fiechter A. 1982; Convenient procedure for the isolation of highly enriched, cytochrome P-450-containing microsomal fraction from Candida tropicalis. Analytical Biochemistry 126:179–182
    [Google Scholar]
  23. Kärenlampi S.O, Marin E., Hännien O.O.P. 1980; Occurrence of cytochrome P-450 in yeasts. Journal of General Microbiology 120:529–533
    [Google Scholar]
  24. Large P.J. 1981; Microbial growth on methylated amines. In Microbial Growth on C1 Compounds. Proceedings of Third International Symposium pp. 55–69 Dalton H. Edited by London: Heyden & Son;
    [Google Scholar]
  25. Large P.J., Green J. 1984; Oxidation of mono-, di-and trimethylamine by methazotrophic yeasts: properties of the microsomal and peroxisomal enzymes involved and comparison with bacterial systems. In Microbial Growth on C1 Compounds. Proceedings of Fourth International Symposium. Crawford R.L. Edited by Washington: American Society for Microbiology; (in the Press)
    [Google Scholar]
  26. Large P.J., Mcdougall H. 1975; An enzymic method for the microestimation of trimethylamine. Analytical Biochemistry 64:304–310
    [Google Scholar]
  27. Large P.J., Eady R.R., Murden D.J. 1969; An enzymic method for the microestimation of methyl-amine, ethylamine, and n-propylamine. Analytical Biochemistry 32:402–407
    [Google Scholar]
  28. Lloyd D. 1974 The Mitochondria of Micro-organisms pp. 64–81 London: Academic Press;
    [Google Scholar]
  29. Meiberg J.B.M., Harder W. 1978; Aerobic and anaerobic metabolism of trimethylamine, dimethylamine and methylamine in Hyphomicrobium X. Journal of General Microbiology 106:265–276
    [Google Scholar]
  30. Nash T. 1953; The colorimetric estimation of formaldehyde by means of the Hantzsch reaction. Biochemical Journal 55:416–421
    [Google Scholar]
  31. Osumi M., Miwa N., Teranishi Y., Tanaka A., Fukui S. 1975; Development of microbodies in Candida tropicalis during incubation in an-alkane medium. Archives of Microbiology 103:1–11
    [Google Scholar]
  32. Pettit F.H., Orme-Johnson W., Ziegler D.M. 1964; The requirement for flavin adenine dinucleotide by a liver microsomal oxygenase catalyzing the oxidation of alkylaryl amines. Biochemical and Biophysical Research Communications 16:444–448
    [Google Scholar]
  33. Robinson J., Cooper J.M. 1970; Method of determining oxygen concentrations in biological media, suitable for calibration of the oxygen electrode. Analytical Biochemistry 33:390–399
    [Google Scholar]
  34. Sahm H., Roggenkamp R., Wagner F., Hinkelma-Mann W. 1975; Microbodies in methanol-grown Candida boidinii. Journal of General Microbiology 88:218–222
    [Google Scholar]
  35. Sawicki E., Hauser T.R., Stanley T.W., Elbert W. 1961; The 3-methyl-2-benzothiazolone hydrazone test. Sensitive new methods for the detection, rapid estimation and determination of aliphatic aldehydes. Analytical Chemistry 33:93–96
    [Google Scholar]
  36. Shaltiel S., Er-EL Z. 1973; Hydrophobic chromatography: use for purification of glycogen synthetase. Proceedings of the National Academy of Sciences of the United States of America 70:778–781
    [Google Scholar]
  37. Tanaka A., Yasuhara S., Kawamoto S., Fukui S., Osumi M. 1976; Development of microbodies in the yeast Kloeckera growing on methanol. Journal of Bacteriology 126:919–927
    [Google Scholar]
  38. Tipton K.F. 1980; Monoamine oxidase. In Enzymatic Basis of Detoxication 1 pp. 355–370 Jakoby W.B. Edited by New York: Academic Press;
    [Google Scholar]
  39. Van Dijken J.P., Bos P. 1981; Utilization of amines by yeasts. Archives of Microbiology 128:320–324
    [Google Scholar]
  40. Veenhuis M., Van Dijken J.P., Harder W. 1980; A new method for the cytochemical demonstration of phosphatase activities in yeasts based on the use of cerous ions. FEMS Microbiology Letters 9:285–291
    [Google Scholar]
  41. Veenhuis M., Van Dijken J.P., Harder W. 1983; The significance of peroxisomes in the metabolism of one-carbon compounds in yeasts. Advances in Microbial Physiology 24:1–82
    [Google Scholar]
  42. Yamada H., Kishimoto N., Kumagai H. 1976; Metabolism of N-substituted amines by yeasts. Journal of Fermentation Technology 54:726–737
    [Google Scholar]
  43. Yamada T., Tanaka A., Fukui S. 1982; Properties of catalase purified from whole cells and peroxisomes of n-alkane-grown Candida tropicalis. European Journal of Biochemistry 125:517–521
    [Google Scholar]
  44. Zatman L.J. 1981; A search for patterns in methylotrophic pathways.In Microbial Growth on C1Compounds. Proceedings of Third International Symposium pp. 42–54 Dalton H. Edited by London: Hcydcn & Son;
    [Google Scholar]
  45. Zwart K.B., Harder W. 1983; Regulation of the metabolism of some alkylated amines in the yeasts Candida utilis and Hansenula polymorpha. Journal of General Microbiology 129:3157–3169
    [Google Scholar]
  46. Zwart K., Veenhuis M., Van Dijken J.P., Harder W. 1980; Development of amine-oxidase-containing peroxisomes in yeasts during growth on glucose in the presence of mcthylamine as the sole source of nitrogen. Archives of Microbiology 126:117–126
    [Google Scholar]
  47. Zwart K.B., Veenhuis M., Plat G., Harder W. 1983; Characterization of glyoxysomes in yeast and their transformation into peroxisomes in response to changes in environmental conditions. Archives of Microbiology 136:28–38
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-130-10-2577
Loading
/content/journal/micro/10.1099/00221287-130-10-2577
Loading

Data & Media loading...

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