Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1973 Jan;113(1):218–223. doi: 10.1128/jb.113.1.218-223.1973

Betaine-Homocysteine Transmethylase in Pseudomonas denitrificans, a Vitamin B12 Overproducer

Raymond F White 1, Louis Kaplan 1, Jerome Birnbaum 1
PMCID: PMC251621  PMID: 4688138

Abstract

A pantothenate-methionine auxotroph (J741) of Pseudomonas denitrificans was isolated whose growth requirement for methionine could not be satisfied by known precursors of the amino acid, including homocysteine. However, some “methyl rich” compounds such as betaine and dimethylacetothetin (DMT) could satisfy the requirement. S-Methyl-methionine and S-adenosylmethionine were ineffective. Extracts were found to contain an enzyme, betaine-homocysteine transmethylase (BHTase), that uses betaine or DMT as a methyl donor and homocysteine as an acceptor to produce methionine. Growth of J741 in methionine leads to a total repression of the BHTase, whereas the use of DMT leads to a three- to sixfold stimulation of enzyme synthesis compared to betaine-grown cells. The pantothenate requirement is unrelated to the methionine auxotrophy, since the growth of other single auxotrophic mutants as well as revertants of J741 still have their methionine requirement satisfied by betaine or DMT. Another methionine auxotroph that could not use betaine for growth was devoid of BHTase activity.

Full text

PDF
223

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. BRAY R. C., SHEMIN D. On the biosynthesis of vitamin B-12. The derivation of the corrin structure from 5-aminolevulinic acid and the methyl group of methionine. J Biol Chem. 1963 Apr;238:1501–1508. [PubMed] [Google Scholar]
  2. BRAY R., SHEMIN D. The biosynthesis of the porphyrin-like moiety of vitamin B12. II. The origin of the methyl groups. Biochim Biophys Acta. 1958 Dec;30(3):647–648. doi: 10.1016/0006-3002(58)90120-3. [DOI] [PubMed] [Google Scholar]
  3. Balish E., Shapiro S. K. Cystathionine as a precursor of methionine in Escherichia coli and Aerobacter aerogenes. J Bacteriol. 1966 Nov;92(5):1331–1336. doi: 10.1128/jb.92.5.1331-1336.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cauthen S. E., Foster M. A., Woods D. D. Methionine synthesis by extracts of Salmonella typhimurium. Biochem J. 1966 Feb;98(2):630–635. doi: 10.1042/bj0980630. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Demain A. L., Daniels H. J., Schnable L., White R. F. Specificity of the stimulatory effect of betaine on the vitamin B12 fermentation. Nature. 1968 Dec 28;220(5174):1324–1325. doi: 10.1038/2201324a0. [DOI] [PubMed] [Google Scholar]
  6. Demain A. L., White R. F. Porphyrin overproduction by Pseudomonas denitrificans: essentiality of betaine and stimulation by ethionine. J Bacteriol. 1971 Aug;107(2):456–460. doi: 10.1128/jb.107.2.456-460.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. KENSLER C. J., LANGEMANN H. Metabolism of choline and related compounds by hepatic tissue from several species including man. Proc Soc Exp Biol Med. 1954 Feb;85(2):364–367. doi: 10.3181/00379727-85-20882. [DOI] [PubMed] [Google Scholar]
  8. KLEE W. A. The synthesis of methionine by enzymic transmethylation. IX. Formation of subunits of thetin-homocys-teine methylpherase by reaction with sodium dodecyl sulfate. Biochim Biophys Acta. 1962 Jun 4;59:562–568. doi: 10.1016/0006-3002(62)90634-0. [DOI] [PubMed] [Google Scholar]
  9. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  10. Lago B. D., Demain A. L. Alternate requirement for vitamin B12 or methionine in mutants of Pseudomonas denitrificans, a vitamin B12-producing bacterium. J Bacteriol. 1969 Jul;99(1):347–349. doi: 10.1128/jb.99.1.347-349.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. MANDEL L. R., BOREK E. Variability in the structure of ribonucleic acid. Biochem Biophys Res Commun. 1961 Jan 25;4:14–18. doi: 10.1016/0006-291x(61)90246-7. [DOI] [PubMed] [Google Scholar]
  12. MORNINGSTAR J. F., KISLIUK R. L. INTERRELATIONS BETWEEN TWO PATHWAYS OF METHIONINE BIOSYNTHESIS IN AEROBACTER AEROGENES. J Gen Microbiol. 1965 Apr;39:43–51. doi: 10.1099/00221287-39-1-43. [DOI] [PubMed] [Google Scholar]
  13. Mangum J. H., North J. A. Vitamin B 12-dependent methionine biosynthesis in HEp-2 cells. Biochem Biophys Res Commun. 1968 Jul 11;32(1):105–110. doi: 10.1016/0006-291x(68)90433-6. [DOI] [PubMed] [Google Scholar]
  14. Mardon D. N., Balish E. Methionine biosynthesis in Candida albicans. I. S-adenosyl-L-methionine (or S-methyl-L-methionine): homocysteine methyltransferase in cell-free extracts from yeast-like cells. Can J Microbiol. 1971 Jun;17(6):795–802. doi: 10.1139/m71-126. [DOI] [PubMed] [Google Scholar]
  15. SHAPIRO S. K. Adenosylmethioninehomocysteine transmethylase. Biochim Biophys Acta. 1958 Aug;29(2):405–409. doi: 10.1016/0006-3002(58)90199-9. [DOI] [PubMed] [Google Scholar]
  16. SHAPIRO S. K., YPHANTIS D. A., ALMENAS A. BIOSYNTHESIS OF METHIONINE IN SACCHAROMYCES CEREVISIAE. PARTIAL PURIFICATION AND PROPERTIES OF S-ADENOSYLMETHIONINE: HOMOCYSTEINE METHYLTRANSFERASE. J Biol Chem. 1964 May;239:1551–1556. [PubMed] [Google Scholar]
  17. SHAPIRO S. K., YPHANTIS D. A. Assay of S-methylmethionine and S-adenosylmethionine homocysteine transmethylases. Biochim Biophys Acta. 1959 Nov;36:241–244. doi: 10.1016/0006-3002(59)90089-7. [DOI] [PubMed] [Google Scholar]
  18. WILLIAMS J. N., Jr, MONSON W. J., SREENIVASAN A., DIETRICH L. S., HARPER A. E., ELVEHJEM C. A. Effects of a vitamin B12 deficiency on liver enzymes in the rat. J Biol Chem. 1953 May;202(1):151–156. [PubMed] [Google Scholar]
  19. White R. F., Demain A. L. Catabolism of betaine and its relationship to cobalamin overproduction. Biochim Biophys Acta. 1971 Apr 20;237(1):112–119. doi: 10.1016/0304-4165(71)90036-5. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES