A linear megaplasmid, p1CP, carrying the genes for chlorocatechol catabolism of Rhodococcus opacus 1CP

König, Christina; Eulberg, Dirk; Gröning, Janosch A. D.; Lakner, Silvia; Seibert, Volker; Kaschabek, Stefan R.; Schlömann, Michael

doi:10.1099/mic.0.27217-0

Cited by 42 publications

(16 citation statements)

References 49 publications

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“…Most remarkably, it had been shown that certain enzymes must have evolved divergently to those ones typical for proteobacteria providing functional convergence (Eulberg et al ., ; Moiseeva et al ., ). Genes for the 3‐chlorocatechol‐catabolic pathway have been identified to be located on a 740‐kb large linear megaplasmid p1CP (König et al ., ). With the exception of a benzoate‐catabolic gene cluster (Bauer, ), little is known on enzyme activities of peripheral degradation, their genetic localization and on their degree of redundancy in strain 1CP.…”

Section: Introductionmentioning

confidence: 97%

Gene redundancy of two-component (chloro)phenol hydroxylases inRhodococcus opacus1CP

Gröning

Eulberg

Tischler

et al. 2014

FEMS Microbiol Lett

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Among other factors, a distinct gene redundancy is discussed to facilitate high metabolic versatility of rhodococci. Rhodococcus opacus 1CP is a typical member in that respect and degrades a multitude of (chlorinated) aromatic compounds. In contrast to the central pathways of aromatic degradation in strain 1CP, little is known about the degree of gene redundancy and to what extent this is reflected on protein level within the steps of peripheral degradation. By means of degenerated primers deduced from tryptic peptides of a purified phenol hydroxylase component and using the amplified fragment as a labelled probe against genomic 1CP-DNA, three gene sets encoding three different two-component phenol hydroxylases pheA1/pheA2(1-3) could be identified. One of them was found to be located on the megaplasmid p1CP, which confirms the role of these elements for metabolic versatility. Protein chromatography of phenol- and 4-chlorophenol-grown 1CP-biomass gave first evidences on a functional expression of these oxygenases, which could be initially characterised in respect of their substrate specificity.

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Section: Introductionmentioning

confidence: 97%

Gene redundancy of two-component (chloro)phenol hydroxylases inRhodococcus opacus1CP

Gröning

Eulberg

Tischler

et al. 2014

FEMS Microbiol Lett

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“…Digoxigenin (DIG) labeling, blotting, hybridization, and detection procedures were performed as described previously (29). A 688-bp BamHI/SpeI fragment of plasmid p1CPS1 served as a template for probe preparation, and genomic 1CP DNA was digested with the endonucleases ApaI, BamHI, EcoRI, and PstI.…”

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confidence: 99%

Identification of a Novel Self-Sufficient Styrene Monooxygenase from Rhodococcus opacus 1CP

et al. 2009

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Sequence analysis of a 9-kb genomic fragment of the actinobacterium Rhodococcus opacus 1CP led to identification of an open reading frame encoding a novel fusion protein, StyA2B, with a putative function in styrene metabolism via styrene oxide and phenylacetic acid. Gene cluster analysis indicated that the highly related fusion proteins of Nocardia farcinica IFM10152 and Arthrobacter aurescens TC1 are involved in a similar physiological process. Whereas 413 amino acids of the N terminus of StyA2B are highly similar to those of the oxygenases of two-component styrene monooxygenases (SMOs) from pseudomonads, the residual 160 amino acids of the C terminus show significant homology to the flavin reductases of these systems. Cloning and functional expression of His 10 -StyA2B revealed for the first time that the fusion protein does in fact catalyze two separate reactions. Strictly NADH-dependent reduction of flavins and highly enantioselective oxygenation of styrene to (S)-styrene oxide were shown. Inhibition studies and photometric analysis of recombinant StyA2B indicated the absence of tightly bound heme and flavin cofactors in this self-sufficient monooxygenase. StyA2B oxygenates a spectrum of aromatic compounds similar to those of two-component SMOs. However, the specific activities of the flavin-reducing and styrene-oxidizing functions of StyA2B are one to two orders of magnitude lower than those of StyA/StyB from Pseudomonas sp. strain VLB120.The incorporation of one atom of oxygen during hydroxylation, epoxidation, sulfoxidation, or Baeyer-Villiger oxidation is a common initial step of the aerobic degradation of aromatic compounds by microorganisms. In bacteria, these reactions are most frequently catalyzed by inducible flavoprotein monooxygenases (EC 1.14.13 [57]). The majority of these enzymes (socalled single-component flavoprotein monooxygenases) utilize electrons from NAD(P)H, which are transferred to a noncovalently bound flavin adenine dinucleotide (FAD) in order to activate molecular oxygen as a flavin (hydro)peroxide. Depending on the protonation of this intermediate and the type of substrate, an oxygen atom is then incorporated by nucleophilic or electrophilic attack. More recently, different twocomponent flavoprotein monooxygenases have been characterized (57). These systems cover an NAD(P)H-dependent flavin reductase in order to generate reduced flavin and an oxygenase that utilizes this cofactor for the activation of oxygen.The exquisite regio-and stereoselectivities of oxygen insertion by flavoprotein monooxygenases favor these enzymes for biocatalytic applications (23,24,33). This is especially true because chemical synthesis approaches by hetero-or homogenic catalysis often do not yield a sufficiently high enantiomeric excess for the production of pharmaceuticals and their chiral building blocks. The use of oxygen as an inexpensive nontoxic oxidant and mild reaction conditions are additional advantages with the potential for increasing the environmental sustainability of oxygenase-catalyzed bio...

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“…For detection of extra chromosomal DNA, plasmid isolation and pulse field gel electrophoresis assays were developed as described by König and colleagues (). Yeast Chromosome PFG marker (New England Biolabs, IZASA, Barcelona, Spain) was used as reference.…”

Section: Methodsmentioning

confidence: 99%

Combination of degradation pathways for naphthalene utilization in Rhodococcus sp. strain TFB

Tomás-Gallardo

Gómez-Álvarez

Santero

et al. 2013

Microbial Biotechnology

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Rhodococcus sp. strain TFB is a metabolic versatile bacterium able to grow on naphthalene as the only carbon and energy source. Applying proteomic, genetic and biochemical approaches, we propose in this paper that, at least, three coordinated but independently regulated set of genes are combined to degrade naphthalene in TFB. First, proteins involved in tetralin degradation are also induced by naphthalene and may carry out its conversion to salicylaldehyde. This is the only part of the naphthalene degradation pathway showing glucose catabolite repression. Second, a salicylaldehyde dehydrogenase activity that converts salicylaldehyde to salicylate is detected in naphthalene-grown cells but not in tetralin-or salicylate-grown cells. Finally, we describe the chromosomally located nag genes, encoding the gentisate pathway for salicylate conversion into fumarate and pyruvate, which are only induced by salicylate and not by naphthalene. This work shows how biodegradation pathways in Rhodococcus sp. strain TFB could be assembled using elements from different pathways mainly because of the laxity of the regulatory systems and the broad specificity of the catabolic enzymes.

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A linear megaplasmid, p1CP, carrying the genes for chlorocatechol catabolism of Rhodococcus opacus 1CP

Cited by 42 publications

References 49 publications

Gene redundancy of two-component (chloro)phenol hydroxylases inRhodococcus opacus1CP

Gene redundancy of two-component (chloro)phenol hydroxylases inRhodococcus opacus1CP

Identification of a Novel Self-Sufficient Styrene Monooxygenase from Rhodococcus opacus 1CP

Combination of degradation pathways for naphthalene utilization in Rhodococcus sp. strain TFB

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