Gene Cluster on pAO1 of
            <i>Arthrobacter nicotinovorans</i>
            Involved in Degradation of the Plant Alkaloid Nicotine: Cloning, Purification, and Characterization of 2,6-Dihydroxypyridine 3-Hydroxylase

Baitsch, Daniel; Sandu, Cristinel; Brandsch, Roderich; Igloi, Gabor L.

doi:10.1128/jb.183.18.5262-5267.2001

Cited by 77 publications

(66 citation statements)

References 34 publications

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“…2,6-dihydroxypyridine hydroxylase of Arthrobacter nicotinovorans (Baitsch et al, 2001; GenBank accession no. AF373840) with 32 % identical positions, and various salicylate hydroxylases, for example, one from Pseudomonas putida strain PpG7 [You et al, 1991; GenBank accession no.…”

Section: Preliminary Characterization Of Orfs Neighbouring the Maleylmentioning

confidence: 99%

Characterization of a gene cluster encoding the maleylacetate reductase from Ralstonia eutropha 335T, an enzyme recruited for growth with 4-fluorobenzoate

et al. 2004

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A gene cluster containing a gene for maleylacetate reductase (EC 1.3.1.32) was cloned from Ralstonia eutropha 335 T (DSM 531 T ), which is able to utilize 4-fluorobenzoate as sole carbon source. Sequencing of this gene cluster showed that the R. eutropha 335 T maleylacetate reductase gene, macA, is part of a novel gene cluster, which is not related to the known maleylacetate-reductase-encoding gene clusters. It otherwise comprises a gene for a hypothetical membrane transport protein, macB, possibly co-transcribed with macA, and a presumed regulatory gene, macR, which is divergently transcribed from macBA. MacA was found to be most closely related to TftE, the maleylacetate reductase from Burkholderia cepacia AC1100 (62 % identical positions) and to a presumed maleylacetate reductase from a dinitrotoluene catabolic gene cluster from B. cepacia R34 (61 % identical positions). By expressing macA in Escherichia coli, it was confirmed that macA encodes a functional maleylacetate reductase. Purification of maleylacetate reductase from 4-fluorobenzoate-grown R. eutropha 335 T cells allowed determination of the N-terminal sequence of the purified protein, which was shown to be identical to that predicted from the cloned macA gene, thus proving that the gene is, in fact, recruited for growth of R. eutropha 335 T with this substrate. INTRODUCTIONMaleylacetate reductases (EC 1.3.1.32) play a crucial role in the aerobic microbial degradation of aromatic compounds. They catalyse the NADH-or NADPH-dependent reduction of maleylacetate or 2-chloromaleylacetate to 3-oxoadipate ( Fig. 1) or of substituted maleylacetates to substituted 3-oxoadipates. In fungi, maleylacetate reductases contribute to the catabolism of very common substrates, such as tyrosine, gentisate, benzoate, 4-hydroxybenzoate, protocatechuate, vanillate, resorcinol and phenol (Karasevich & Ivoilov, 1977;Buswell & Eriksson, 1979;Gaal & Neujahr, 1979;Sparnins et al., 1979;Anderson & Dagley, 1980;Jones et al., 1995). For bacteria, it has been shown that maleylacetate reductases are involved in the degradation of resorcinol and 2,4-dihydroxybenzoate via hydroxyquinol (Larway & Evans, 1965;Chapman & Ribbons, 1976;Stolz & Knackmuss, 1993). Other substrates whose catabolic routes comprise this activity have a more complex structure, such as 2-hydroxydibenzo-p-dioxin and 3-hydroxydibenzofuran (Armengaud et al., 1999), or carry unusual substituents, such as nitro or sulfo groups (Spain & Gibson, 1991;Feigel & Knackmuss, 1993;Jain et al., 1994;Rani & Lalithakumari, 1994), fluorine or chlorine atoms (Duxbury et al., 1970; Schlömann et al., 1990a;Latus et al., 1995;Zaborina et al., 1995;Daubaras et al., 1996;Miyauchi et al., 1999).The maleylacetate reductase genes characterized so far tend to belong to specialized gene clusters for the degradation of aromatic compounds. Thus, 3-hydroxydibenzofuran and 2-hydroxydibenzo-p-dioxin degradation via hydroxyquinol appears to be encoded by a specialized gene cluster comprising most of the genes for the pathway including a maleylac...

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Section: Preliminary Characterization Of Orfs Neighbouring the Maleylmentioning

confidence: 99%

Characterization of a gene cluster encoding the maleylacetate reductase from Ralstonia eutropha 335T, an enzyme recruited for growth with 4-fluorobenzoate

et al. 2004

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“…The identification of 2,6-dihydroxypyridine (DHP) and ␥-N-methylaminobutyrate indicated, however, that DHPON was the physiological intermediate in nicotine degradation. The subsequent enzymatic steps involved in the turnover of DHP into 2,3,6-trihydroxypyridine by an FAD-dependent hydroxylase (2,13) and the novel flavoenzyme ␥-N-methylaminobutyrate oxidase (MABO) responsible for the oxidative demethylation of ␥-N-methylaminobutyrate (4) have been described.…”

mentioning

confidence: 99%

“…Genes known or presumed to be involved in nicotine catabolism are clustered on the megaplasmid pAO1 of A. nicotinovorans (2,15). They are expressed in the presence of nicotine in the growth medium and form a nicotine regulon.…”

mentioning

confidence: 99%

An α/β-Fold C—C Bond Hydrolase Is Involved in a Central Step of Nicotine Catabolism by Arthrobacter nicotinovorans

Sachelaru¹,

Schiltz²,

Igloi

et al. 2005

J Bacteriol

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“…Succinic semialdehyde ([M-H] Ϫ peak at m/z 101) was produced during enzymatic degradation, which was verified with the commercial standard by LC-MS analysis. Generally, hydroxylase is a group of enzymes that catalyze oxidation reactions in which one of the two atoms of molecular oxygen is incorporated into the substrate and the other is used to oxidize NADH or NADPH (1,19,24). NADH was necessary for the cleavage of HSP, and this mechanism was similar to Hirschberg's and Nakano's reports.…”

Section: Discussionmentioning

confidence: 51%

A Novel Gene, Encoding 6-Hydroxy-3-Succinoylpyridine Hydroxylase, Involved in Nicotine Degradation by Pseudomonas putida Strain S16

Tang

Wang

et al. 2008

Appl Environ Microbiol

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Previous research suggested that Pseudomonas spp. may attack the pyrrolidine ring of nicotine in a way similar to mammalian metabolism, resulting in the formation of pseudooxynicotine, the direct precursor of a potent tobacco-specific lung carcinogen. In addition, the subsequent intermediates, 6-hydroxy-3-succinoylpyridine (HSP) and 2,5-dihydroxypyridine (DHP) in the Pseudomonas nicotine degradation pathway are two important precursors for drug syntheses. However, there is little information on the molecular mechanism for nicotine degradation via the pyrrolidine pathway until now. In this study we cloned and sequenced a 4,879-bp gene cluster involved in nicotine degradation. Intermediates N-methylmyosmine, pseudooxynicotine, 3-succinoylpyridine, HSP, and DHP were identified from resting cell reactions of the transformant containing the gene cluster and shown to be identical to those of the pyrrolidine pathway reported in wild-type strain Pseudomonas putida S16. The gene for 6-hydroxy-3-succinoylpyridine hydroxylase (HSP hydroxylase) catalyzing HSP directly to DHP was cloned, sequenced, and expressed in Escherichia coli, and the purified HSP hydroxylase (38 kDa) is NADH dependent. DNA sequence analysis of this 936-bp fragment reveals that the deduced amino acid shows no similarity with any protein of known function.

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Gene Cluster on pAO1 of Arthrobacter nicotinovorans Involved in Degradation of the Plant Alkaloid Nicotine: Cloning, Purification, and Characterization of 2,6-Dihydroxypyridine 3-Hydroxylase

Cited by 77 publications

References 34 publications

Characterization of a gene cluster encoding the maleylacetate reductase from Ralstonia eutropha 335T, an enzyme recruited for growth with 4-fluorobenzoate

Characterization of a gene cluster encoding the maleylacetate reductase from Ralstonia eutropha 335T, an enzyme recruited for growth with 4-fluorobenzoate

An α/β-Fold C—C Bond Hydrolase Is Involved in a Central Step of Nicotine Catabolism by Arthrobacter nicotinovorans

A Novel Gene, Encoding 6-Hydroxy-3-Succinoylpyridine Hydroxylase, Involved in Nicotine Degradation by Pseudomonas putida Strain S16

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