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Application of nitroarene dioxygenases in the design of novel strains that degrade chloronitrobenzenes.

Ju KS, Parales RE - Microb Biotechnol (2009)

Bottom Line: To address this need, we created engineered strains with a novel degradation pathway that reduces the total number of steps required to convert chloronitrobenzenes into compounds of central metabolism.We examined the ability of 2-nitrotoluene 2,3-dioxygenase from Acidovorax sp. strain JS42, nitrobenzene 1,2-dioxygenase (NBDO) from Comamonas sp. strain JS765, as well as active-site mutants of NBDO to generate chlorocatechols from chloronitrobenzenes, and identified the most efficient enzymes.Introduction of the wild-type NBDO and the F293Q variant into Ralstonia sp. strain JS705, a strain carrying the modified ortho pathway for chlorocatechol metabolism, resulted in bacterial strains that were able to sustainably grow on all three chloronitrobenzene isomers without addition of co-substrates or co-inducers.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, College of Biological Sciences, University of California, Davis, CA 95616, USA.

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Nitrite released from CNBs by E. coli strains expressing wild‐type and mutant nitroarene dioxygenases. 2CNB, white bars; 3CNB, grey bars; 4CNB, black bars. N = 3; error bars indicate standard deviations.
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f3: Nitrite released from CNBs by E. coli strains expressing wild‐type and mutant nitroarene dioxygenases. 2CNB, white bars; 3CNB, grey bars; 4CNB, black bars. N = 3; error bars indicate standard deviations.

Mentions: The activity of the nitroarene dioxygenases towards the different CNBs and their ability to form chlorocatechols was determined as a function of the amount of nitrite released at the end of 6 h biotransformation reactions (Fig. 3). NBDO had comparable activities with 2CNB and 4CNB, but had the highest activity with 3CNB. In contrast, 2NTDO produced approximately twice as much product from 2CNB compared with NBDO, but its average activities with 3CNB and 4CNB were only 4% and < 1% of the NBDO activities, respectively.


Application of nitroarene dioxygenases in the design of novel strains that degrade chloronitrobenzenes.

Ju KS, Parales RE - Microb Biotechnol (2009)

Nitrite released from CNBs by E. coli strains expressing wild‐type and mutant nitroarene dioxygenases. 2CNB, white bars; 3CNB, grey bars; 4CNB, black bars. N = 3; error bars indicate standard deviations.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3815844&req=5

f3: Nitrite released from CNBs by E. coli strains expressing wild‐type and mutant nitroarene dioxygenases. 2CNB, white bars; 3CNB, grey bars; 4CNB, black bars. N = 3; error bars indicate standard deviations.
Mentions: The activity of the nitroarene dioxygenases towards the different CNBs and their ability to form chlorocatechols was determined as a function of the amount of nitrite released at the end of 6 h biotransformation reactions (Fig. 3). NBDO had comparable activities with 2CNB and 4CNB, but had the highest activity with 3CNB. In contrast, 2NTDO produced approximately twice as much product from 2CNB compared with NBDO, but its average activities with 3CNB and 4CNB were only 4% and < 1% of the NBDO activities, respectively.

Bottom Line: To address this need, we created engineered strains with a novel degradation pathway that reduces the total number of steps required to convert chloronitrobenzenes into compounds of central metabolism.We examined the ability of 2-nitrotoluene 2,3-dioxygenase from Acidovorax sp. strain JS42, nitrobenzene 1,2-dioxygenase (NBDO) from Comamonas sp. strain JS765, as well as active-site mutants of NBDO to generate chlorocatechols from chloronitrobenzenes, and identified the most efficient enzymes.Introduction of the wild-type NBDO and the F293Q variant into Ralstonia sp. strain JS705, a strain carrying the modified ortho pathway for chlorocatechol metabolism, resulted in bacterial strains that were able to sustainably grow on all three chloronitrobenzene isomers without addition of co-substrates or co-inducers.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, College of Biological Sciences, University of California, Davis, CA 95616, USA.

Show MeSH
Related in: MedlinePlus