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Metabolism of 2-chloro-4-nitrophenol in a gram negative bacterium, Burkholderia sp. RKJ 800.

Arora PK, Jain RK - PLoS ONE (2012)

Bottom Line: On the basis of thin layer chromatography, high performance liquid chromatography and gas chromatography-mass spectrometry, chlorohydroquinone (CHQ) and hydroquinone (HQ) were identified as major metabolites of the degradation pathway of 2C4NP.Our studies clearly showed that Burkholderia sp.RKJ 800 degraded 2-chloro-4-nitrophenol via hydroquinone pathway.

View Article: PubMed Central - PubMed

Affiliation: Environmental Biotechnology, Institute of Microbial Technology (CSIR), Chandigarh, India. arora484@gmail.com

ABSTRACT
A 2-chloro-4-nitrophenol (2C4NP) degrading bacterial strain designated as RKJ 800 was isolated from a pesticide contaminated site of India by enrichment method and utilized 2C4NP as sole source of carbon and energy. The stoichiometric amounts of nitrite and chloride ions were detected during the degradation of 2C4NP. On the basis of thin layer chromatography, high performance liquid chromatography and gas chromatography-mass spectrometry, chlorohydroquinone (CHQ) and hydroquinone (HQ) were identified as major metabolites of the degradation pathway of 2C4NP. Manganese dependent HQ dioxygenase activity was observed in the crude extract of 2C4NP induced cells of the strain RKJ 800 that suggested the cleavage of the HQ to γ-hydroxymuconic semialdehyde. On the basis of the 16S rRNA gene sequencing, strain RKJ 800 was identified as a member of genus Burkholderia. Our studies clearly showed that Burkholderia sp. RKJ 800 degraded 2-chloro-4-nitrophenol via hydroquinone pathway. The pathway identified in a gram negative bacterium, Burkholderia sp. strain RKJ 800 was differed from previously reported 2C4NP degradation pathway in another gram-negative Burkholderia sp. SJ98. This is the first report of the formation of CHQ and HQ in the degradation of 2C4NP by any gram-negative bacteria. Laboratory-scale soil microcosm studies showed that strain RKJ 800 is a suitable candidate for bioremediation of 2C4NP contaminated sites.

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HPLC analysis showing complete depletion of 2C4NP with appearance of metabolites.
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pone-0038676-g004: HPLC analysis showing complete depletion of 2C4NP with appearance of metabolites.

Mentions: HPLC confirmed complete depletion of 2C4NP by strain RKJ 800 within 48 h (Fig. 4). In the 12 h sample, only parent compound was detected. The metabolite I was detected in the sample of 24 h and 36 h whereas the metabolite II was detected only in the 36 h sample. In the sample of 48 h, neither parent compound nor metabolite was detected. The retention time of 2C4NP, metabolite 1 and metabolite II were 16.3 min, 6.7 min and 4.5 min, respectively. The retention times of metabolite I and II were exactly match of authentic CHQ and HQ.


Metabolism of 2-chloro-4-nitrophenol in a gram negative bacterium, Burkholderia sp. RKJ 800.

Arora PK, Jain RK - PLoS ONE (2012)

HPLC analysis showing complete depletion of 2C4NP with appearance of metabolites.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3368897&req=5

pone-0038676-g004: HPLC analysis showing complete depletion of 2C4NP with appearance of metabolites.
Mentions: HPLC confirmed complete depletion of 2C4NP by strain RKJ 800 within 48 h (Fig. 4). In the 12 h sample, only parent compound was detected. The metabolite I was detected in the sample of 24 h and 36 h whereas the metabolite II was detected only in the 36 h sample. In the sample of 48 h, neither parent compound nor metabolite was detected. The retention time of 2C4NP, metabolite 1 and metabolite II were 16.3 min, 6.7 min and 4.5 min, respectively. The retention times of metabolite I and II were exactly match of authentic CHQ and HQ.

Bottom Line: On the basis of thin layer chromatography, high performance liquid chromatography and gas chromatography-mass spectrometry, chlorohydroquinone (CHQ) and hydroquinone (HQ) were identified as major metabolites of the degradation pathway of 2C4NP.Our studies clearly showed that Burkholderia sp.RKJ 800 degraded 2-chloro-4-nitrophenol via hydroquinone pathway.

View Article: PubMed Central - PubMed

Affiliation: Environmental Biotechnology, Institute of Microbial Technology (CSIR), Chandigarh, India. arora484@gmail.com

ABSTRACT
A 2-chloro-4-nitrophenol (2C4NP) degrading bacterial strain designated as RKJ 800 was isolated from a pesticide contaminated site of India by enrichment method and utilized 2C4NP as sole source of carbon and energy. The stoichiometric amounts of nitrite and chloride ions were detected during the degradation of 2C4NP. On the basis of thin layer chromatography, high performance liquid chromatography and gas chromatography-mass spectrometry, chlorohydroquinone (CHQ) and hydroquinone (HQ) were identified as major metabolites of the degradation pathway of 2C4NP. Manganese dependent HQ dioxygenase activity was observed in the crude extract of 2C4NP induced cells of the strain RKJ 800 that suggested the cleavage of the HQ to γ-hydroxymuconic semialdehyde. On the basis of the 16S rRNA gene sequencing, strain RKJ 800 was identified as a member of genus Burkholderia. Our studies clearly showed that Burkholderia sp. RKJ 800 degraded 2-chloro-4-nitrophenol via hydroquinone pathway. The pathway identified in a gram negative bacterium, Burkholderia sp. strain RKJ 800 was differed from previously reported 2C4NP degradation pathway in another gram-negative Burkholderia sp. SJ98. This is the first report of the formation of CHQ and HQ in the degradation of 2C4NP by any gram-negative bacteria. Laboratory-scale soil microcosm studies showed that strain RKJ 800 is a suitable candidate for bioremediation of 2C4NP contaminated sites.

Show MeSH
Related in: MedlinePlus