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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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TLC analysis showing the degradation of 2C4NP.
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pone-0038676-g003: TLC analysis showing the degradation of 2C4NP.

Mentions: TLC, HPLC and GC-MS studies were carried out to elucidate metabolic pathway of 2C4NP by strain RKJ 800. TLC results indicated depletion of 2C4NP with appearance of the two metabolites (Fig. 3). No metabolite was detected in the sample of 0 h and 12 h. The presence of metabolite I was indicated in the sample of 24 h and 36 h whereas the presence of metabolite II was indicated only in the sample of 36 h. In the sample of 48 h, neither the presence of any metabolite nor parent compound was indicated. The Rf values of metabolite I, II and 2C4NP were 0.64, 0.48 and 0.72 respectively. The Rf values of metabolite I and II exactly matched to that of standard CHQ and HQ respectively. Furthermore, when TLC plates were sprayed with folin ciocalteu’s reagent, an immediate blue coloration was apparent in the case of suspected chlorohydroquinone and hydroquinone.


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

Arora PK, Jain RK - PLoS ONE (2012)

TLC analysis showing the degradation of 2C4NP.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0038676-g003: TLC analysis showing the degradation of 2C4NP.
Mentions: TLC, HPLC and GC-MS studies were carried out to elucidate metabolic pathway of 2C4NP by strain RKJ 800. TLC results indicated depletion of 2C4NP with appearance of the two metabolites (Fig. 3). No metabolite was detected in the sample of 0 h and 12 h. The presence of metabolite I was indicated in the sample of 24 h and 36 h whereas the presence of metabolite II was indicated only in the sample of 36 h. In the sample of 48 h, neither the presence of any metabolite nor parent compound was indicated. The Rf values of metabolite I, II and 2C4NP were 0.64, 0.48 and 0.72 respectively. The Rf values of metabolite I and II exactly matched to that of standard CHQ and HQ respectively. Furthermore, when TLC plates were sprayed with folin ciocalteu’s reagent, an immediate blue coloration was apparent in the case of suspected chlorohydroquinone and hydroquinone.

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