Gene Cluster of Arthrobacter ilicis Rü61a Involved in the Degradation of Quinaldine to Anthranilate
A genetic analysis of the anthranilate pathway of quinaldine degradation was performed. A 23-kb region of DNA from Arthrobacter ilicis Rü61a was cloned into the cosmid pVK100. Although Escherichia coli clones containing the recombinant cosmid did not transform quinaldine, cosmids harboring the 23-k...
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Veröffentlicht in: | The Journal of biological chemistry 2003-07, Vol.278 (30), p.27483 |
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Sprache: | eng |
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Zusammenfassung: | A genetic analysis of the anthranilate pathway of quinaldine degradation
was performed. A 23-kb region of DNA from Arthrobacter ilicis
Rü61a was cloned into the cosmid pVK100. Although Escherichia
coli clones containing the recombinant cosmid did not transform
quinaldine, cosmids harboring the 23-kb region, or a 10.8-kb stretch of this
region, conferred to Pseudomonas putida KT2440 the ability to
cometabolically convert quinaldine to anthranilate. The 10.8-kb fragment thus
contains the genes coding for quinaldine 4-oxidase (Qox),
1 H -4-oxoquinaldine 3-monooxygenase,
1 H -3-hydroxy-4-oxoquinaldine 2,4-dioxygenase, and
N -acetylanthranilate amidase. The qoxLMS genes coding for
the molybdopterin cytosine dinucleotide-(MCD-), FeSI-, FeSII-, and
FAD-containing Qox were inserted into the expression vector pJB653, generating
pKP1. Qox is the first MCD-containing enzyme to be synthesized in a
catalytically fully competent form by a heterologous host, P. putida
KT2440 pKP1; the catalytic properties and the UV-visible and EPR spectra of
Qox purified from P. putida KT2440 pKP1 were essentially like those
of wild-type Qox. This provides a starting point for the construction of
protein variants of Qox by site-directed mutagenesis. Downstream of the
qoxLMS genes, a putative gene whose deduced amino acid sequence
showed 37% similarity to the cofactor-inserting chaperone XdhC was located.
Additional open reading frames identified on the 23-kb segment may encode
further enzymes (a glutamyl tRNA synthetase, an esterase, two short-chain
dehydrogenases/reductases, an ATPase belonging to the AAA family, a
2-hydroxyhepta-2,4-diene-1,7-dioate
isomerase/5-oxopent-3-ene-1,2,5-tricarboxylate decarboxylase-like protein, and
an enzyme of the mandelate racemase group) and hypothetical proteins involved
in transcriptional regulation, and metabolite transport. |
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ISSN: | 0021-9258 1083-351X |
DOI: | 10.1074/jbc.M301330200 |