Development and characterization of a lux-modified 2,4-dichlorophenol-degrading Burkholderia sp RASC

L J Shaw, Y Beaton, L A Glover, K Killham, A A Meharg

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

lux-marked biosensors for assessing the toxicity and bioremediation potential of polluted environments may complement traditional chemical techniques. luxCDABE genes were introduced into the chromosome of the 2,4-dichlorophenol (2,4-DCP)-mineralizing bacterium, Burkholderia sp. RASC c2, by biparental mating using the Tn4431 system. Experiments revealed that light output was constitutive and related to cell biomass concentration during exponential growth. The transposon insertion was stable and did not interrupt 2,4-DCP-degradative genes, and expression of luxCDABE did not constitute a metabolic burden to the cell. A bioluminescence response was detectable at sublethal 2,4-DCP concentrations: at <10.26 mu g ml(-1) bioluminescence was stimulated (e.g. 218% of control), but at concentrations >60 mu g ml(-1) it declined to <1%. Investigating the effect of [C-14]-2,4-DCP concentration on the evolution of (CO2)-C-14 revealed that, for initial concentrations of 2.5-25 mu g ml(-1), approximate to 55% of the added C-14 was mineralized after 24 h compared with <1% at 50 and 100 mu g ml(-1). Inhibition of 2,4-DCP mineralization between 25 and 50 mu g ml(-1) corresponded well to the EC50 value (33.83 mu g ml(-1)) obtained from bioluminescence inhibition studies. lux-marked RASC c2 may therefore be used as a functionally (i.e. 2,4-DCP degrader) and environmentally relevant biosensor of toxicity and biodegradation inhibition.

Original languageEnglish
Pages (from-to)393-399
Number of pages7
JournalEnvironmental Microbiology
Volume1
Publication statusPublished - 1999

Keywords

  • ENERGY-TRANSDUCING MEMBRANES
  • PSEUDOMONAS-FLUORESCENS
  • MODIFIED BACTERIAL
  • PENTACHLOROPHENOL DEGRADATION
  • CELL
  • SOIL
  • BIOAVAILABILITY
  • BIOREMEDIATION
  • CHLOROPHENOLS
  • POLLUTANTS

Cite this

Shaw, L. J., Beaton, Y., Glover, L. A., Killham, K., & Meharg, A. A. (1999). Development and characterization of a lux-modified 2,4-dichlorophenol-degrading Burkholderia sp RASC. Environmental Microbiology, 1, 393-399.

Development and characterization of a lux-modified 2,4-dichlorophenol-degrading Burkholderia sp RASC. / Shaw, L J ; Beaton, Y ; Glover, L A ; Killham, K ; Meharg, A A .

In: Environmental Microbiology, Vol. 1, 1999, p. 393-399.

Research output: Contribution to journalArticle

Shaw, LJ, Beaton, Y, Glover, LA, Killham, K & Meharg, AA 1999, 'Development and characterization of a lux-modified 2,4-dichlorophenol-degrading Burkholderia sp RASC' Environmental Microbiology, vol. 1, pp. 393-399.
Shaw, L J ; Beaton, Y ; Glover, L A ; Killham, K ; Meharg, A A . / Development and characterization of a lux-modified 2,4-dichlorophenol-degrading Burkholderia sp RASC. In: Environmental Microbiology. 1999 ; Vol. 1. pp. 393-399.
@article{bb51d78596ea4816be3a0427376f20b6,
title = "Development and characterization of a lux-modified 2,4-dichlorophenol-degrading Burkholderia sp RASC",
abstract = "lux-marked biosensors for assessing the toxicity and bioremediation potential of polluted environments may complement traditional chemical techniques. luxCDABE genes were introduced into the chromosome of the 2,4-dichlorophenol (2,4-DCP)-mineralizing bacterium, Burkholderia sp. RASC c2, by biparental mating using the Tn4431 system. Experiments revealed that light output was constitutive and related to cell biomass concentration during exponential growth. The transposon insertion was stable and did not interrupt 2,4-DCP-degradative genes, and expression of luxCDABE did not constitute a metabolic burden to the cell. A bioluminescence response was detectable at sublethal 2,4-DCP concentrations: at <10.26 mu g ml(-1) bioluminescence was stimulated (e.g. 218{\%} of control), but at concentrations >60 mu g ml(-1) it declined to <1{\%}. Investigating the effect of [C-14]-2,4-DCP concentration on the evolution of (CO2)-C-14 revealed that, for initial concentrations of 2.5-25 mu g ml(-1), approximate to 55{\%} of the added C-14 was mineralized after 24 h compared with <1{\%} at 50 and 100 mu g ml(-1). Inhibition of 2,4-DCP mineralization between 25 and 50 mu g ml(-1) corresponded well to the EC50 value (33.83 mu g ml(-1)) obtained from bioluminescence inhibition studies. lux-marked RASC c2 may therefore be used as a functionally (i.e. 2,4-DCP degrader) and environmentally relevant biosensor of toxicity and biodegradation inhibition.",
keywords = "ENERGY-TRANSDUCING MEMBRANES, PSEUDOMONAS-FLUORESCENS, MODIFIED BACTERIAL, PENTACHLOROPHENOL DEGRADATION, CELL, SOIL, BIOAVAILABILITY, BIOREMEDIATION, CHLOROPHENOLS, POLLUTANTS",
author = "Shaw, {L J} and Y Beaton and Glover, {L A} and K Killham and Meharg, {A A}",
year = "1999",
language = "English",
volume = "1",
pages = "393--399",
journal = "Environmental Microbiology",
issn = "1462-2912",
publisher = "BLACKWELL PUBLISHING LTD",

}

TY - JOUR

T1 - Development and characterization of a lux-modified 2,4-dichlorophenol-degrading Burkholderia sp RASC

AU - Shaw, L J

AU - Beaton, Y

AU - Glover, L A

AU - Killham, K

AU - Meharg, A A

PY - 1999

Y1 - 1999

N2 - lux-marked biosensors for assessing the toxicity and bioremediation potential of polluted environments may complement traditional chemical techniques. luxCDABE genes were introduced into the chromosome of the 2,4-dichlorophenol (2,4-DCP)-mineralizing bacterium, Burkholderia sp. RASC c2, by biparental mating using the Tn4431 system. Experiments revealed that light output was constitutive and related to cell biomass concentration during exponential growth. The transposon insertion was stable and did not interrupt 2,4-DCP-degradative genes, and expression of luxCDABE did not constitute a metabolic burden to the cell. A bioluminescence response was detectable at sublethal 2,4-DCP concentrations: at <10.26 mu g ml(-1) bioluminescence was stimulated (e.g. 218% of control), but at concentrations >60 mu g ml(-1) it declined to <1%. Investigating the effect of [C-14]-2,4-DCP concentration on the evolution of (CO2)-C-14 revealed that, for initial concentrations of 2.5-25 mu g ml(-1), approximate to 55% of the added C-14 was mineralized after 24 h compared with <1% at 50 and 100 mu g ml(-1). Inhibition of 2,4-DCP mineralization between 25 and 50 mu g ml(-1) corresponded well to the EC50 value (33.83 mu g ml(-1)) obtained from bioluminescence inhibition studies. lux-marked RASC c2 may therefore be used as a functionally (i.e. 2,4-DCP degrader) and environmentally relevant biosensor of toxicity and biodegradation inhibition.

AB - lux-marked biosensors for assessing the toxicity and bioremediation potential of polluted environments may complement traditional chemical techniques. luxCDABE genes were introduced into the chromosome of the 2,4-dichlorophenol (2,4-DCP)-mineralizing bacterium, Burkholderia sp. RASC c2, by biparental mating using the Tn4431 system. Experiments revealed that light output was constitutive and related to cell biomass concentration during exponential growth. The transposon insertion was stable and did not interrupt 2,4-DCP-degradative genes, and expression of luxCDABE did not constitute a metabolic burden to the cell. A bioluminescence response was detectable at sublethal 2,4-DCP concentrations: at <10.26 mu g ml(-1) bioluminescence was stimulated (e.g. 218% of control), but at concentrations >60 mu g ml(-1) it declined to <1%. Investigating the effect of [C-14]-2,4-DCP concentration on the evolution of (CO2)-C-14 revealed that, for initial concentrations of 2.5-25 mu g ml(-1), approximate to 55% of the added C-14 was mineralized after 24 h compared with <1% at 50 and 100 mu g ml(-1). Inhibition of 2,4-DCP mineralization between 25 and 50 mu g ml(-1) corresponded well to the EC50 value (33.83 mu g ml(-1)) obtained from bioluminescence inhibition studies. lux-marked RASC c2 may therefore be used as a functionally (i.e. 2,4-DCP degrader) and environmentally relevant biosensor of toxicity and biodegradation inhibition.

KW - ENERGY-TRANSDUCING MEMBRANES

KW - PSEUDOMONAS-FLUORESCENS

KW - MODIFIED BACTERIAL

KW - PENTACHLOROPHENOL DEGRADATION

KW - CELL

KW - SOIL

KW - BIOAVAILABILITY

KW - BIOREMEDIATION

KW - CHLOROPHENOLS

KW - POLLUTANTS

M3 - Article

VL - 1

SP - 393

EP - 399

JO - Environmental Microbiology

JF - Environmental Microbiology

SN - 1462-2912

ER -