Enzyme recruitment in vitro: use of cloned genes to extend the range of haloaromatics degraded by Pseudomonas sp. strain B13

Lehrbach, P. R.; Zeyer, Josef; Reineke, Walter; Knackmuss, Hans‐Joachim; Timmis, Kenneth N.

doi:10.1128/jb.158.3.1025-1032.1984

Cited by 123 publications

(31 citation statements)

References 28 publications

(31 reference statements)

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“…DNA manipulation and genetic methods Methods for plasmid isolation, transformation, cleavage by restriction enzymes, ligation and agarose gel electrophoresis have previously been described (Maniatis et al, 1982;Franklin et al, 1983). Matings between E.coli and P.putida were performed as described by Lehrbach et al (1984). Polymerase chain reaction amplification of the xylT DNA region was performed by using VentR DNA polymerase (Biolabs).…”

Section: Methodsmentioning

confidence: 99%

In vivo reactivation of catechol 2,3-dioxygenase mediated by a chloroplast-type ferredoxin: a bacterial strategy to expand the substrate specificity of aromatic degradative pathways.

Polissi¹,

Harayama²

1993

The EMBO Journal

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The meta‐cleavage operon of the TOL plasmid pWW0 of Pseudomonas putida contains 13 genes responsible for the oxidation of benzoate and toluates to Krebs cycle intermediates via estradiol (meta) cleavage of (methyl)catechol. The functions of all the genes are known with the exception of xylT. We constructed pWW0 mutants defective in the xylT gene, and found that these mutants were not able to grow on p‐toluate while they were still capable of growing on benzoate and m‐toluate. In the xylT mutants, all the meta‐cleavage enzymes were induced by p‐toluate with the exception of catechol 2,3‐dioxygenase whose activity was 1% of the p‐toluate‐induced activity in wild‐type cells. Addition of 4‐methylcatechol to m‐toluate‐grown wild‐type and xylT cells resulted in the inactivation of catechol 2,3‐dioxygenase in these cells. In the wild‐type strain but not in the xylT mutant, the catechol 2,3‐dioxygenase activity was regenerated in a short time. The regeneration of the catechol 2,3‐dioxygenase activity was also observed in H2O2‐treated wild‐type cells, but not in H2O2‐treated xylT cells. We concluded that the xylT product is required for the regeneration of catechol 2,3‐dioxygenase.

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Section: Methodsmentioning

confidence: 99%

In vivo reactivation of catechol 2,3-dioxygenase mediated by a chloroplast-type ferredoxin: a bacterial strategy to expand the substrate specificity of aromatic degradative pathways.

Polissi¹,

Harayama²

1993

The EMBO Journal

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“…This type of a patchwork assembly has subsequently been used successfully to construct strains with a widened range of degradative abilities or improved degradation rates [3,6,367]. The development of in vitro cloning procedures offers a powerful tool in constructing novel strains that can degrade various environmental pollutants [376]. However, as Reineke [367] pointed out, the construction of hybrid pathways is limited to preexisting genetic material found in laboratory strains, while conventional enrichment techniques can use the vast and unexploited reservoir of genetic material available in nature.…”

Section: Genetic Basis Of Degradation and Construction Of Novel Strainsmentioning

confidence: 99%

Microbial breakdown of halogenated aromatic pesticides and related compounds

Häggblom

1992

FEMS Microbiology Letters

363

132

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Considerable progress has been made in the last few years in understanding the mechanisms of microbial degradation of halogenated aromatic compounds. Much is already known about the degradation mechanisms under aerobic conditions, and metabolism under anaerobiosis has lately received increasing attention. The removal of the halogen substituent is a key step in degradation of halogenated aromatics. This may occur as an initial step via reductive, hydrolytic or oxygenolytic mechanisms, or after cleavage of the aromatic ring at a later stage of metabolism. In addition to degradation, several biotransformation reactions, such as methylation and polymerization, may take place and produce more toxic or recalcitrant metabolites. Studies with pure bacterial and fungal cultures have given detailed information on the biodegradation pathways of several halogenated aromatic compounds. Several of the key enzymes have been purified or studied in cell extracts, and there is an increasing understanding of the organization and regulation of the genes involved in haloaromatic degradation. This review will focus on the biodegradation and biotransformation pathways that have been established for halogenated phenols, phenoxyalkanoic acids, benzoic acids, benzenes, anilines and structurally related halogenated aromatic pesticides. There is a growing interest in developing microbiological methods for clean‐up of soil and water contaminated with halogenated aromatic compounds.

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“…On the other hand, the TOL xylS and xylXYZ genes have been used to expand the range of haloaromatics degraded by Pseudomonas sp. strain B13 (6,12,24,25).…”

mentioning

confidence: 99%

A substrate-dependent biological containment system for Pseudomonas putida based on the Escherichia coli gef gene

Jensen

Ramos

Kaneva

et al. 1993

Appl Environ Microbiol

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A model substrate-dependent suicide system to biologically contain Pseudomonas putida KT2440 is reported. The system consists of two elements. One element carries a fusion between a synthetic bc promoter (PA-4w03) and the gefgene, which encodes a killing function. This element is contained within a transposaseless mini-TnS transposon so that it can be integrated at random locations on the Pseudomonas chromosome. The second element, harbored by plasmid pCC102, is designed to control the first and bears a fusion between the promoter of the P. putida TOL plasmid-encoded meta-cleavage pathway operon (Pm) and the lacI gene, encoding the Lac repressor, plus xyLS2, coding for a positive regulator of Pm. In liquid culture under optimal growth conditions and in sterile and nonsterile soil microcosms, P. putida KT2440(pWWO) bearing the containment system behaves as designed. In the presence of a XylS effector, such as m-methylbenzoate, the LacI protein is synthesized, preventing the expression of the killing function. In the absence of effectors, expression of the PA,4w03::gef cassette is no longer prevented and a high rate of cell killing is observed. Fluctuation test analyses revealed that mutants resistant to cell killing arise at a frequency of around 10'S to 10-6 per cell per

show abstract

Enzyme recruitment in vitro: use of cloned genes to extend the range of haloaromatics degraded by Pseudomonas sp. strain B13

Cited by 123 publications

References 28 publications

In vivo reactivation of catechol 2,3-dioxygenase mediated by a chloroplast-type ferredoxin: a bacterial strategy to expand the substrate specificity of aromatic degradative pathways.

In vivo reactivation of catechol 2,3-dioxygenase mediated by a chloroplast-type ferredoxin: a bacterial strategy to expand the substrate specificity of aromatic degradative pathways.

Microbial breakdown of halogenated aromatic pesticides and related compounds

A substrate-dependent biological containment system for Pseudomonas putida based on the Escherichia coli gef gene

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