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PMID: 1917841 Published · ppublish English Journal Article

Genetic analysis, nucleotide sequence, and products of two Pseudomonas denitrificans cob genes encoding nicotinate-nucleotide: dimethylbenzimidazole phosphoribosyltransferase and cobalamin (5'-phosphate) synthase.

Journal of bacteriology ·Vol. 173 ·No. 19 ·1991-10-00 ·Pages 6066-73

Cameron B, Blanche F, Rouyez MC, Bisch D, Famechon A, Couder M, Cauchois L, Thibaut D, Debussche L, Crouzet J

Abstract

Tn5 Sp(r) transposons have been inserted into the 8-kb Pseudomonas denitrificans DNA fragment from complementation group D, which carries cob genes. Genetic analysis and the nucleotide sequence revealed that only two cob genes (cobU and cobV) were found on this cob genomic locus. Nicotinate-nucleotide: dimethylbenzimidazole phosphoribosyltransferase (EC 2.4.2.21) was assayed and purified to homogeneity from a P. denitrificans strain in which cobU and cobV were amplified. The purified enzyme was identified as the cobU gene product on the basis of identical molecular weights and N-terminal sequences. Cobalamin (5'-phosphate) synthase activity was increased when cobV was amplified in P. denitrificans. The partially purified enzyme catalyzed not only the synthesis of cobalamin 5'-phosphate from GDP-cobinamide and alpha-ribazole 5'-phosphate but also the one-step synthesis of cobalamin from GDP-cobinamide and alpha-ribazole. Biochemical data provided evidence that cobV encodes cobalamin (5'-phosphate) synthase.

Related Genes

cob cobU cobV

MeSH Terms

Amino Acid Sequence Bacterial Proteins Base Composition Base Sequence Cobamides/biosynthesis,genetics DNA, Bacterial/chemistry,isolation & purification Deoxyribonuclease EcoRI Deoxyribonuclease HindIII Gene Amplification Genes, Bacterial Genetic Complementation Test Kinetics Molecular Sequence Data Molecular Weight Multienzyme Complexes Nitrogenous Group Transferases Nucleotidyltransferases Pentosyltransferases/genetics Pseudomonas/enzymology,genetics Transferases/genetics

Chemicals

Bacterial Proteins Cobamides DNA, Bacterial Multienzyme Complexes Transferases Pentosyltransferases nicotinate-nucleotide-dimethylbenzimidazole phosphoribosyltransferase Nitrogenous Group Transferases CobV protein, Pseudomonas denitrificans Nucleotidyltransferases Deoxyribonuclease EcoRI Deoxyribonuclease HindIII

Authors & Affiliations

10 authors, click to expand affiliations / ORCID

Cameron B

Unité de Biologie Moléculaire, Institut des Biotechnologies, Vitry-sur-Seine, France.

Blanche F

Rouyez M C

Bisch D

Famechon A

Couder M

Cauchois L

Thibaut D

Debussche L

Crouzet J

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Article Info

Journal

Journal of bacteriology

Abbr.

J Bacteriol

ISSN

0021-9193

DOI

10.1128/jb.173.19.6066-6073.1991

Published

1991-10-00

Pages

6066-73

Language

English

Region

United States

NLM ID

2985120R

PMCID

PMC208353

Subset

Databases

GENBANK

M62868, M64126, M64127, M64128, M64129, S52988, S52991, S53821, S57155, X61007

External Links

PubMed source DOI: 10.1128/jb.173.19.6066-6073.1991 PMC Full Text View journal details

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Genetic analysis, nucleotide sequence, and products of two Pseudomonas denitrificans cob genes encoding nicotinate-nucleotide: dimethylbenzimidazole phosphoribosyltransferase and cobalamin (5'-phosphate) synthase.

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