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. 1992 Nov;174(22):7452–7454. doi: 10.1128/jb.174.22.7452-7454.1992

Purification and characterization of Cob(II)yrinic acid a,c-diamide reductase from Pseudomonas denitrificans.

F Blanche 1, L Maton 1, L Debussche 1, D Thibaut 1
PMCID: PMC207442  PMID: 1429467

Abstract

An NADH-dependent flavoenzyme exhibiting cob(II)yrinic acid a,c-diamide reductase activity was purified 6,300-fold to homogeneity from Pseudomonas denitrificans and sequenced at its N terminus. This enzyme of the cobalamin biosynthetic pathway reduced to the Co(I) state all of the Co(II)-corrinoids isolated from this microorganism.

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Selected References

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

  1. Blanche F., Couder M., Debussche L., Thibaut D., Cameron B., Crouzet J. Biosynthesis of vitamin B12: stepwise amidation of carboxyl groups b, d, e, and g of cobyrinic acid a,c-diamide is catalyzed by one enzyme in Pseudomonas denitrificans. J Bacteriol. 1991 Oct;173(19):6046–6051. doi: 10.1128/jb.173.19.6046-6051.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Blanche F., Debussche L., Famechon A., Thibaut D., Cameron B., Crouzet J. A bifunctional protein from Pseudomonas denitrificans carries cobinamide kinase and cobinamide phosphate guanylyltransferase activities. J Bacteriol. 1991 Oct;173(19):6052–6057. doi: 10.1128/jb.173.19.6052-6057.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Blanche F., Debussche L., Thibaut D., Crouzet J., Cameron B. Purification and characterization of S-adenosyl-L-methionine: uroporphyrinogen III methyltransferase from Pseudomonas denitrificans. J Bacteriol. 1989 Aug;171(8):4222–4231. doi: 10.1128/jb.171.8.4222-4231.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cameron B., Blanche F., Rouyez M. C., Bisch D., Famechon A., Couder M., Cauchois L., Thibaut D., Debussche L., Crouzet J. Genetic analysis, nucleotide sequence, and products of two Pseudomonas denitrificans cob genes encoding nicotinate-nucleotide: dimethylbenzimidazole phosphoribosyltransferase and cobalamin (5'-phosphate) synthase. J Bacteriol. 1991 Oct;173(19):6066–6073. doi: 10.1128/jb.173.19.6066-6073.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cameron B., Briggs K., Pridmore S., Brefort G., Crouzet J. Cloning and analysis of genes involved in coenzyme B12 biosynthesis in Pseudomonas denitrificans. J Bacteriol. 1989 Jan;171(1):547–557. doi: 10.1128/jb.171.1.547-557.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cameron B., Guilhot C., Blanche F., Cauchois L., Rouyez M. C., Rigault S., Levy-Schil S., Crouzet J. Genetic and sequence analyses of a Pseudomonas denitrificans DNA fragment containing two cob genes. J Bacteriol. 1991 Oct;173(19):6058–6065. doi: 10.1128/jb.173.19.6058-6065.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Crouzet J., Cameron B., Cauchois L., Rigault S., Rouyez M. C., Blanche F., Thibaut D., Debussche L. Genetic and sequence analysis of an 8.7-kilobase Pseudomonas denitrificans fragment carrying eight genes involved in transformation of precorrin-2 to cobyrinic acid. J Bacteriol. 1990 Oct;172(10):5980–5990. doi: 10.1128/jb.172.10.5980-5990.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Crouzet J., Cauchois L., Blanche F., Debussche L., Thibaut D., Rouyez M. C., Rigault S., Mayaux J. F., Cameron B. Nucleotide sequence of a Pseudomonas denitrificans 5.4-kilobase DNA fragment containing five cob genes and identification of structural genes encoding S-adenosyl-L-methionine: uroporphyrinogen III methyltransferase and cobyrinic acid a,c-diamide synthase. J Bacteriol. 1990 Oct;172(10):5968–5979. doi: 10.1128/jb.172.10.5968-5979.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Crouzet J., Levy-Schil S., Cameron B., Cauchois L., Rigault S., Rouyez M. C., Blanche F., Debussche L., Thibaut D. Nucleotide sequence and genetic analysis of a 13.1-kilobase-pair Pseudomonas denitrificans DNA fragment containing five cob genes and identification of structural genes encoding Cob(I)alamin adenosyltransferase, cobyric acid synthase, and bifunctional cobinamide kinase-cobinamide phosphate guanylyltransferase. J Bacteriol. 1991 Oct;173(19):6074–6087. doi: 10.1128/jb.173.19.6074-6087.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Debussche L., Couder M., Thibaut D., Cameron B., Crouzet J., Blanche F. Assay, purification, and characterization of cobaltochelatase, a unique complex enzyme catalyzing cobalt insertion in hydrogenobyrinic acid a,c-diamide during coenzyme B12 biosynthesis in Pseudomonas denitrificans. J Bacteriol. 1992 Nov;174(22):7445–7451. doi: 10.1128/jb.174.22.7445-7451.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Debussche L., Couder M., Thibaut D., Cameron B., Crouzet J., Blanche F. Purification and partial characterization of Cob(I)alamin adenosyltransferase from Pseudomonas denitrificans. J Bacteriol. 1991 Oct;173(19):6300–6302. doi: 10.1128/jb.173.19.6300-6302.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Debussche L., Thibaut D., Cameron B., Crouzet J., Blanche F. Purification and characterization of cobyrinic acid a,c-diamide synthase from Pseudomonas denitrificans. J Bacteriol. 1990 Nov;172(11):6239–6244. doi: 10.1128/jb.172.11.6239-6244.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Djalali M., Gueant J. L., Lambert D., el Kholty S., Saunier M., Nicolas J. P. High-performance liquid chromatographic separation and dual competitive binding assay of corrinoids in biological material. J Chromatogr. 1990 Jul 13;529(1):81–91. doi: 10.1016/s0378-4347(00)83809-8. [DOI] [PubMed] [Google Scholar]
  14. Fenton W. A., Rosenberg L. E. The defect in the cbl B class of human methylmalonic acidemia: deficiency of cob(I)alamin adenosyltransferase activity in extracts of cultured fibroblasts. Biochem Biophys Res Commun. 1981 Jan 15;98(1):283–289. doi: 10.1016/0006-291x(81)91900-8. [DOI] [PubMed] [Google Scholar]
  15. Schrauzer G. N., Deutsch E., Windgassen R. J. The nucleophilicity of vitamin B12. J Am Chem Soc. 1968 Apr 24;90(9):2441–2442. doi: 10.1021/ja01011a054. [DOI] [PubMed] [Google Scholar]
  16. Vitols E., Walker G. A., Huennekens F. M. Enzymatic conversion of vitamin B-12s to a cobamide coenzyme, alpha-(5,6-dimethylbenzimidazolyl)deoxyadenosylcobamide (adenosyl-B-12). J Biol Chem. 1966 Apr 10;241(7):1455–1461. [PubMed] [Google Scholar]
  17. Walker G. A., Murphy S., Huennekens F. M. Enzymatic conversion of vitamin B 12a to adenosyl-B 12: evidence for the existence of two separate reducing systems. Arch Biochem Biophys. 1969 Oct;134(1):95–102. doi: 10.1016/0003-9861(69)90255-0. [DOI] [PubMed] [Google Scholar]

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