Cloning and characterization of the genes encoding nitrilotriacetate monooxygenase of Chelatobacter heintzii ATCC 29600.

Détails

ID Serval
serval:BIB_F2895798DBD0
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Titre
Cloning and characterization of the genes encoding nitrilotriacetate monooxygenase of Chelatobacter heintzii ATCC 29600.
Périodique
Journal of Bacteriology
Auteur(s)
Knobel H.R., Egli T., van der Meer J.R.
ISSN
0021-9193 (Print)
ISSN-L
0021-9193
Statut éditorial
Publié
Date de publication
1996
Peer-reviewed
Oui
Volume
178
Numéro
21
Pages
6123-6132
Langue
anglais
Résumé
A 6.2-kb DNA fragment containing the genes for the nitrilotriacetate (NTA) monooxygenase of Chelatobacter heintzii ATCC 29600 was cloned and characterized by DNA sequencing and expression studies. The nucleotide sequence contained three major open reading frames (ORFs). Two of the ORFs, which were oriented divergently with an intergenic region of 307 bp, could be assigned to the NTA monooxygenase components A and B. The predicted N-terminal amino acid sequences of these ORFs were identical with those determined for the purified components. We therefore named these genes ntaA (for component A of NTA monooxygenase) and ntaB (for component B). The ntaA and ntaB genes could be expressed in Escherichia coli DH5alpha, and the gene products were visualized after Western blotting (immunoblotting) and incubation with polyclonal antibodies against component A or B. By mixing overproduced NtaB from E. coli and purified component A from C. heintzii ATCC 29600, reconstitution of a functional NTA monooxygenase complex was possible. The deduced gene product of ntaA showed only significant homology to SoxA (involved in dibenzothiophene degradation) and to SnaA (involved in pristamycin synthesis); that of ntaB shared weak homologies in one domain with other NADH:flavine mononucleotide oxidoreductases. These homologies provide no conclusive answer as to the possible evolutionary origin of the NTA monooxygenase. The deduced gene product of the third ORF (ORF1) had homology in the N-terminal region with the GntR class of bacterial regulator proteins and therefore may encode a regulator protein, possibly involved in regulation of ntaA and ntaB expression.
Mots-clé
Amino Acid Sequence, Base Sequence, Cloning, Molecular, DNA, Bacterial, Escherichia coli/metabolism, Gene Expression, Gram-Negative Aerobic Bacteria/enzymology, Gram-Negative Aerobic Bacteria/genetics, Mixed Function Oxygenases/classification, Mixed Function Oxygenases/genetics, Molecular Sequence Data, Molecular Structure, Phylogeny, Sequence Analysis, DNA, Sequence Homology, Amino Acid
Pubmed
Web of science
Création de la notice
21/01/2008 14:36
Dernière modification de la notice
03/03/2018 22:40
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