Degradation of trichloroethylene by Pseudomonas cepacia G4 and the constitutive mutant strain G4 5223 PR1 in aquifer microcosms

Krumme, M.L.; Timmis, Kenneth N.; Dwyer, Daryl F.

doi:10.1128/aem.59.8.2746-2749.1993

Cited by 53 publications

(24 citation statements)

References 17 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Trichloroethylene (TCE), cis ‐1,2‐dichloroethylene ( cis ‐DCE) and other chlorinated ethenes constitute a large group of priority environmental pollutants (Krumme et al ., 1993; Bradley and Chapelle, 1998). Remediation of sites contaminated with these compounds is critical because chlorinated ethenes are toxic (Coleman et al ., 2002a), and natural anaerobic degradation often leads to even more toxic compounds such as vinyl chloride, a well‐known carcinogen (Coleman et al ., 2002b).…”

Section: Introductionmentioning

confidence: 99%

“…Toluene ortho ‐monooxygenase (TOM) of Burkholderia cepacia G4 was originally studied for its ability to oxidize TCE (Nelson et al ., 1986). In addition to TCE, TOM can also degrade other chlorinated ethenes such as cis ‐DCE and trans ‐DCE (Shim and Wood, 2000; Canada et al ., 2002); hence, there is considerable interest in using TOM for bioremediation (Krumme et al ., 1993; Winkler et al ., 1995). TOM‐Green, a DNA shuffling variant from our laboratory, has an enhanced degradation rate for TCE (2.5 ± 0.3 versus 1.39 ± 0.05 nmol min −1 mg −1 protein at 67 µM initial liquid concentration), 1,1‐dichloroethylene and trans ‐DCE (Canada et al ., 2002).…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Metabolic pathway engineering to enhance aerobic degradation of chlorinated ethenes and to reduce their toxicity by cloning a novel glutathione S‐transferase, an evolved toluene o‐monooxygenase, and γ‐glutamylcysteine synthetase

Rui¹,

Kwon²,

Reardon³

et al. 2004

Environmental Microbiology

View full text Add to dashboard Cite

Aerobic, co-metabolic bioremediation of trichloroethylene (TCE), cis-1,2-dichloroethylene (cis-DCE) and other chlorinated ethenes with monooxygenase-expressing microorganisms is limited by the toxic epoxides produced as intermediates. A recombinant Escherichia coli strain less sensitive to the toxic effects of cis-DCE, TCE and trans-1,2-dichloroethylene (trans-DCE) degradation has been created by engineering a novel pathway consisting of eight genes including a DNA-shuffled toluene ortho-monooxygenase from Burkholderia cepacia G4 (TOM-Green), a newly discovered glutathione S-transferase (GST) from RhodococcusAD45 (IsoILR1), found to have activity towards epoxypropane and cis-DCE epoxide, and an overexpressed E. coli mutant gamma-glutamylcysteine synthetase (GSHI*). Along with IsoILR1, another new RhodococcusAD45 GST, IsoILR2, was cloned that lacks activity towards cis-DCE epoxide and differs from IsoILR1 by nine amino acids. The recombinant strain in which TOM-Green and IsoILR1 were co-expressed on separate plasmids degraded 1.9-fold more cis-DCE compared with a strain that lacked IsoILR1. In the presence of IsoILR1 and TOM-Green, the addition of GSH1* resulted in a sevenfold increase in the intracellular GSH concentration and a 3.5-fold improvement in the cis-DCE degradation rate based on chloride released (2.1 +/- 0.1 versus 0.6 +/- 0.1 nmol min(-1) mg(-1) protein at 540 microM), a 1.8-fold improvement in the trans-DCE degradation rate (1.29 +/- 0.03 versus 0.71 +/- 0.04 nmol x min(-1) mg(-1) protein at 345 microM) and a 1.7-fold improvement in the TCE degradation rate (6.8 +/- 0.24 versus 4.1 +/- 0.16 nmol x min(-1) mg(-1) protein at 339 microM). For cis-DCE degradation with TOM-Green (based on substrate depletion), V(max) was 27 nmol x min(-1) mg(-1) protein with both IsoILR1 and GSHI* expressed compared with V(max) = 10 nmol x min(-1) mg(-1) protein for the GST(-)GSHI*(-) strain. In addition, cells expressing IsoILR1 and GSHI* grew 78% faster in rich medium than a strain lacking these two heterologous genes.

show abstract

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Metabolic pathway engineering to enhance aerobic degradation of chlorinated ethenes and to reduce their toxicity by cloning a novel glutathione S‐transferase, an evolved toluene o‐monooxygenase, and γ‐glutamylcysteine synthetase

Rui¹,

Kwon²,

Reardon³

et al. 2004

Environmental Microbiology

View full text Add to dashboard Cite

show abstract

“…cepacia was originally described as a saprophyte that causes soft rot of onions (Burkholder, 1950), and was initially considered to be a plant pathogen that is also widely distributed in soil and water (Bergan, 1981). Its metabolic capacities include antagonizing soil-borne plant pathogens (McLoughlin et aL, 1992;Bevivino et aL, 1994) and degrading complex hydrocarbons and xenobiotics (Lessie and Gaffney, 1986;Krumme et aL, 1993;Masson et aL, 1993). It is a common pathogen for opportunistic and nosocomial infections (Martone et aL, 1987;Neu, 1993;Yamagishi et aL, 1993;Lacy et aL, 1994).…”

Section: Introductionmentioning

confidence: 99%

A physical genome map of the Burkholderia cepacia type strain

Rodley

Römling²,

Tümmler

1995

Molecular Microbiology

View full text Add to dashboard Cite

Burkholderia cepacia (basonym Pseudomonas cepacia), the type species of the new genus Burkholderia, is of interest, not only because of its broad catabolic capacity and its ability to antagonize soil-borne plant pathogens, but also because of its causative role in infections in man, which are particularly evident in patients with cystic fibrosis. A physical map of the 8.1 Mb genome of the B. cepacia type-strain ATCC 25416 was constructed by applying two-dimensional pulsed-field gel electrophoresis techniques. Placed onto the macrorestriction map were 38 SpeI, 11 SwaI, 11 PacI, 11 PmeI and six I-CeuI sites, resulting in an average resolution of 105 kbp. Random single-hit linearization by irradiation and restriction mapping uncovered the presence of four circular replicons of 3.65 Mb, 3.17 Mb, 1.07 Mb and 200 kbp in size. The largest replicon harbours four rrn operons while the other two Megabase-size replicons each contain a single rrn operon, suggesting that the genome has three chromosomes and a large plasmid. Within the beta subdivision of proteobacteria, the existence of multiple replicons is not confined to B. cepacia. The phylogenetically related species Burkholderia glumae, Burkholderia pickettii, Burkholderia solanacearum, Alcaligenes eutrophus and the so far unassigned Pseudomonas glathei were also found to harbour more than one Megabase-size replicon.

show abstract

“…The constitutive mutant PR1 has great potential for bioaugmentation of groundwater aquifers, where the toxic nature of phenol and toluene would preclude their environmental use for induction of TCE degradation. In sterile aquifer groundwater and sediment microcosms the constitutive mutant PR1 was found to degrade significant amounts of TCE over a few weeks without addition of nutrients or inducers (Krumme et al 1993). Sixty-three different TCEdegraders were identified in a study of phenol and toluene degrading microbial populations from an aquifer, but none was as effective as strain G4 (Fries et al 1997).…”

Section: Trichloroethylene Biodegradationmentioning

confidence: 99%