Structural analysis and molybdenum‐dependent expression of the pAO1‐encoded nicotine dehydrogenase genes of <i>Arthrobacter nicotinovorans</i>

Grether‐Beck, Susanne; Igloi, Gabor L.; Pust, S.; Schilz, Emil; Decker, Karl; Brandsch, Roderich

doi:10.1111/j.1365-2958.1994.tb00484.x

Cited by 69 publications

(76 citation statements)

References 24 publications

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“…CoxM, CoxS, and CoxL (Fig. 2) and their equivalents in nicotine dehydrogenase of Arthrobacter nicotinovorans (8) exhibited similarities of 74% (32% identity), 83% (42% identity), and 71% (33% identity), respectively. CoxS is 74% similar (41% identical) to aa 1 to 166 of MOP from Desulfovibrio gigas (36).…”

Section: Coxs Contains Motifs Indicative Of Type I and Ii [2fe-2s]mentioning

confidence: 99%

Molecular characterization of the gene cluster coxMSL encoding the molybdenum-containing carbon monoxide dehydrogenase of Oligotropha carboxidovorans

et al. 1995

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The CO dehydrogenase structural genes (cox) and orf4 are clustered in the transcriptional order coxM3 coxS3 coxL3 orf4 on the 128-kb megaplasmid pHCG3 of the carboxidotroph Oligotropha carboxidovorans OM5. Sequence analysis suggested association of molybdopterin cytosine dinucleotide and flavin adenine dinucleotide with CoxL and of the [2Fe-2S] clusters with CoxS.The molybdenum-containing iron-sulfur flavoprotein CODH (26) is the key enzyme in the chemolithoautotrophic utilization of CO by Oligotropha carboxidovorans OM5 (27,29). The large, medium, and small subunits of CODH are encoded by the structural genes coxL, coxM, and coxS, respectively (27). They reside on the 128-kb megaplasmid pHCG3 (20,21). We report here the structural characteristics of the cox genes and the analysis of the deduced amino acid sequences.Abbreviations. The following terms have been abbreviated in this report (abbreviations are shown in parentheses): carbon monoxide dehydrogenase (CODH), aldehyde oxidoreductase (MOP), xanthine dehydrogenase (XDH), flavin adenine dinucleotide (FAD), molybdopterin cytosine dinucleotide (MCD), and amino acid (aa).The strains employed were O. carboxidovorans OM5 (DSM 1227 [29]), Escherichia coli DH5␣ (15), and E. coli K38 (32).The basic recombinant DNA techniques used followed standard protocols (3,25). Plasmids were isolated as described previously (4, 21). The coxS probe (oligonucleotide S) was the 64-fold degenerate 17-mer ATRTGNGCYTTNGCCAT derived from the N-terminal protein sequence MAKAHI of CoxS (12,20). The coxM probe (oligonucleotide M) was the 128-fold degenerate 17-mer ATNCKRTGRTARTCRAA derived from the protein sequence FDYHRI of CoxM (12,20). Deoxyoligonucleotides were 3Ј end labeled with digoxigenin-11-ddUTP. Restriction fragment gene probes were labeled with digoxigenin-11-dUTP by the use of random primers (7). With the 1.45-kb BamHI-EcoRV fragment of pHCG3 (Fig. 1), hybridizations were carried out at 60ЊC and washes were carried out with 1ϫ SSC (1ϫ SSC is 0.15 M NaCl plus 0.015 M sodium citrate) (containing 0.1% sodium dodecyl sulfate) at 68ЊC.The 4.86-kb BamHI-HindIII fragment of pHCG3 carrying the structural genes coxMSL was inserted into the vector phagemid pBluescript I KSϩ (Stratagene, Heidelberg, Germany), yielding phagemid pCDH1 (Fig. 1). Sets of nested deletions were introduced bidirectionally into the cloned region by treatment with exonuclease III-S1 nuclease (10) of the Erasea-Base kit (Promega, Madison, Wis.). Sequencing of both strands of cloned DNA with the primers T3 and T7 (Stratagene, La Jolla, Calif.) was performed with the Taq DyeDeoxy Terminator Cycle Sequencing kit in the model 373A DNA sequencing system (Applied Biosystems, Foster City, Calif.). Each nucleotide position was determined with a redundancy of four. The HUSAR program package (Deutsches Krebsforschungszentrum, Heidelberg, Germany) was employed for sequence analysis.DNA sequence and molecular organization of the coxMSL gene cluster. Southern hybridizations with the oligonucleotides M and S identified the 5Ј ...

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Section: Coxs Contains Motifs Indicative Of Type I and Ii [2fe-2s]mentioning

confidence: 99%

Molecular characterization of the gene cluster coxMSL encoding the molybdenum-containing carbon monoxide dehydrogenase of Oligotropha carboxidovorans

et al. 1995

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“…Insertion of an antibiotic resistance cassette into an open reading frame divergently transcribed from the gene encoding 4-OHBen-CoA ligase, the first enzyme of 4-OHBen degradation, resulted in the generation of a mutant that was unable to grow on 4-OHBen. Sequencing of this region revealed three open reading frames with sequence similarities to bacterial hydroxylating enzymes (21,37,43) as well as to eukaryotic xanthine dehydrogenases (1,28). The work described here suggests that these genes encode the R. palustris 4-OHbenzoyl-CoA reductase (dehydroxylating) and that 4-OHBen is obligatorily metabolized via benzoyl-CoA when it is used as a carbon source for phototrophic growth.…”

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confidence: 99%

“…Several prokaryotic enzymes belonging to this family are nicotine dehydrogenase from Arthrobacter nicotinovorans (ndhABC), carbon dioxide dehydrogenase from Oligotropha carboxidovorans (coxSLM), and carbon monoxide dehydrogenase from Pseudomonas thermocarboxydovorans (cutABC) (21,37,43). These enzymes consist of three subunits similar in size to HbaB, HbaC, and HbaD.…”

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confidence: 99%

4-hydroxybenzoyl coenzyme A reductase (dehydroxylating) is required for anaerobic degradation of 4-hydroxybenzoate by Rhodopseudomonas palustris and shares features with molybdenum-containing hydroxylases

1997

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The anaerobic degradation of 4-hydroxybenzoate is initiated by the formation of 4-hydroxybenzoyl coenzyme A, with the next step proposed to be a dehydroxylation to benzoyl coenzyme A, the starting compound for a central pathway of aromatic compound ring reduction and cleavage. Three open reading frames, divergently transcribed from the 4-hydroxybenzoate coenzyme A ligase gene, hbaA, were identified and sequenced from the phototrophic bacterium Rhodopseudomonas palustris. These genes, named hbaBCD, specify polypeptides of 17.5, 82.6, and 34.5 kDa, respectively. The deduced amino acid sequences show considerable similarities to a group of hydroxylating enzymes involved in CO, xanthine, and nicotine metabolism that have conserved binding sites for [2Fe-2S] clusters and a molybdenum cofactor. Cassette disruption of the hbaB gene yielded a mutant that was unable to grow anaerobically on 4-hydroxybenzoate but grew normally on benzoate. The hbaB mutant cells did not accumulate [ 14 C]benzoyl coenzyme A during short-term uptake of [ 14 C]4-hydroxybenzoate, but benzoyl coenzyme A was the major radioactive metabolite formed by the wild type. In addition, crude extracts of the mutant failed to convert 4-hydroxybenzoyl coenzyme A to benzoyl coenzyme A. This evidence indicates that the hbaBCD genes encode the subunits of a 4-hydroxybenzoyl coenzyme A reductase (dehydroxylating). The sizes of the specified polypeptides are similar to those reported for 4-hydroxybenzoyl coenzyme A reductase isolated from the denitrifying bacterium Thauera aromatica. The amino acid consensus sequence for a molybdenum cofactor binding site is in HbaC. This cofactor appears to be an essential component because anaerobic growth of R. palustris on 4-hydroxybenzoate, but not on benzoate, was retarded unless 0.1 M molybdate was added to the medium. Neither tungstate nor vanadate replaced molybdate, and tungstate competitively inhibited growth stimulation by molybdate.

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“…The genes encoding these enzymes are located on the 160-kb catabolic plasmid pAO1, and the ability to degrade and grow on nicotine is lost when the plasmid is cured from the bacterium (5). The genes ndh and 6hlno have been cloned and sequenced (16,31). Of the kdh genes, the gene of the small [Fe-S] subunit and of the medium FAD subunit have been cloned and sequenced (30).…”

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confidence: 99%

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

Baitsch¹,

Sandu²,

Brandsch³

et al. 2001

J Bacteriol

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The gram-positive soil bacterium Arthrobacter nicotinovorans (formerly known as Arthrobacter oxidans and reclassified by Kodama et al. [22]) has the ability to use nicotine as its sole carbon and energy source (8,10,15,17). Nicotine, the alkaloid of the tobacco plant, is synthesized as the L-isomer, and the first enzyme to attack L-nicotine is a trimeric, molybdopterin cofactor (MoCo) (most probably in its dinucleotide form [18])-, flavin adenine dinucleotide (FAD)-, and [Fe-S] clustercontaining nicotine dehydrogenase (NDH), which hydroxylates the pyridine ring at position 6 (Fig.

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Structural analysis and molybdenum‐dependent expression of the pAO1‐encoded nicotine dehydrogenase genes of Arthrobacter nicotinovorans

Cited by 69 publications

References 24 publications

Molecular characterization of the gene cluster coxMSL encoding the molybdenum-containing carbon monoxide dehydrogenase of Oligotropha carboxidovorans

Molecular characterization of the gene cluster coxMSL encoding the molybdenum-containing carbon monoxide dehydrogenase of Oligotropha carboxidovorans

4-hydroxybenzoyl coenzyme A reductase (dehydroxylating) is required for anaerobic degradation of 4-hydroxybenzoate by Rhodopseudomonas palustris and shares features with molybdenum-containing hydroxylases

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

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