Skip to main content
SpringerLink
Log in

Isolation of pyrene degrading Achromobacter xylooxidans and characterization of metabolic product

  • Original Paper
  • Published:
World Journal of Microbiology and Biotechnology Aims and scope Submit manuscript

Abstract

Polycyclic aromatic hydrocarbons (PAHs) are common ubiquitous pollutants existing in nature with high recalcitrance and toxicity. In this study a bacterium capable of aerobic degradation of high molecular weight PAHs (with special reference to pyrene) was isolated by selective enrichment culture technique from oil refinery effluent sludge. The isolate was characterized as Achromobacter xylooxidans by 16S rRNA gene sequence analysis technique. For the first time it is hereby reported a bacterium capable of effectively degrading pyrene (up to 80%), as evident by reverse phase high performance liquid chromatographic analysis (RP-HPLC). After incubation of Achromobacter xylooxidans in minimal salt medium (MSM) containing pyrene, at concentration of 200 mg/L, as sole source of carbon and energy, there was decrease in pyrene concentration concomitant with increase in bacterial cell protein concentration. RP-HPLC analysis revealed that pyrene was degraded into three metabolites viz. I, II and III. The RP-HPLC eluent fraction were collected from 2.5 to 32.0 min by repeated injection of degraded sample, concentrated and analyzed on gas chromatography mass spectroscopy (GC-MS) for metabolite identification. The fraction shows 1-hydroxypyrene, 1-hydroxy-6-methoxypyrene and 1,6dimethoxypyrene as metabolic product of pyrene degradation, on the basis of their m/z values. On contrary to the reported PAH degradation with reference to pyrene by different isolates till date; the efficient degradation, as evident by RP-HPLC, by this isolate holds a promising potential for planning of bioremediation strategies of contaminated sites.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  • Bogan BW, Lahner LM, Sullivan WR, Paterek JR (2003) Degradation of straight-chain aliphatic and high-molecular-weight polycyclic aromatic hydrocarbons by a strain of Mycobacterium austroafricanum. J Appl Microbiol 94:230–239

    Article  CAS  Google Scholar 

  • Brezna B, Kweon O, Stingley RL, Freeman JP, Khan AA, Polek B, Jones RC, Cerniglia CE (2006) Molecular characterization of cytochrome P450 genes in the polycyclic aromatic hydrocarbon degrading Mycobacterium vanbaalenii PYR-1. Appl Microbiol Biotechnol 71:522–532

    Article  CAS  Google Scholar 

  • Buratti M, Pellegrino O, Brambilla G, Colombi A (2000) Urinary excretion of 1-hydroxypyrene as a biomarker of exposure to polycyclic aromatic hydrocarbons from different sources. Biomarkers 5:368–372

    Article  CAS  Google Scholar 

  • Cerniglia CE (1992) Biodegradation of polycyclic aromatic hydrocarbons. Biodegradation 3:351–368

    Article  CAS  Google Scholar 

  • Costera A, Feidt C, Dziurla MA, Monteau F, Bizec BL, Rychen G (2009) Bioavailability of polycyclic aromatic hydrocarbons (PAHs) from coil and hay matrices in lactating goats. J Agric Food Chem 57:5352–5357

    Article  CAS  Google Scholar 

  • Datta D, Samanta TB (1988) Effects of inducers on metabolism of benzo[a]pyrene in vivo and in vitro: analysis by high pressure liquid chromatography. Biochem Biophys Res Comm 155:493–502

    Article  CAS  Google Scholar 

  • Hamamura N, Olson SH, Ward DM, Inskeep WP (2006) Microbial population dynamics associated with crude oil biodegradation in diverse soils. Appl Environ Microbiol 72:6316–6324

    Article  CAS  Google Scholar 

  • Heitkamp MA, Freeman JP, Miller DW, Cerniglia CE (1988) Pyrene degradation by a Mycobacterium sp.: identification of ring oxidation and ring fission products. Appl Environ Microbiol 54:2556–2565

    CAS  Google Scholar 

  • Kanaly RA, Harayama S (2000) Biodegradation of high-molecular weight polycyclic aromatic hydrocarbons by bacteria. J Bacteriol 182:2059–2067

    Article  CAS  Google Scholar 

  • Kasuga K, Habe H, Chung JS, Yoshida T, Nojiri H, Yamane H, Omori T (2001) Isolation and characterization of the genes encoding a novel oxygenase component of angular dioxygenase from the gram-positive dibenzofuran-degrader Terrabacter sp. strain DBF63. Biochem Biophys Res Commun 283:195–204

    Article  CAS  Google Scholar 

  • Kazunga C, Aitken MD (2000) Products from the incomplete metabolism of pyrene by polycyclic aromatic hydrocarbon degrading bacteria. Appl Environ Microbiol 66:1917–1922

    Article  CAS  Google Scholar 

  • Kim YH, Moody JD, Freeman JP, Brezna B, Engesser KH, Cerniglia CE (2004) Evidence for the existence of PAH-quinone reductase and catechol- O-methyltransferase in Mycobacterium Vanbaalenii PYR-1. J Ind Microbiol Biotechnol 31:507–516

    Article  CAS  Google Scholar 

  • Kim SJ, Kweon O, Jones RC, Edmondson RD, Cerniglia CE (2007) Complete and integrated pyrene degradation pathway in Mycobacterium vanbaalenii PYR-1 based on system biology. J Bacteriol 189:464–472

    Article  CAS  Google Scholar 

  • Lowry OH, Roserough NJ, Farr AL, Randall RJ (1951) Protein measurement with folin phenol reagent. J Biol Chem 193:265

    CAS  Google Scholar 

  • Malik A, Singh KP, Mohan D, Patel DK (2004) Distribution of polycyclic aromatic hydrocarbons in Gomti river system, India. Bull Environ Contam Toxicol 72:1211–1218

    Article  CAS  Google Scholar 

  • Molina M, Araujo R, Hodson RE (1999) Cross-induction of Pyrene and phenanthrene in a Mycobacterium sp. isolated from polycyclic aromatic hydrocarbon contaminated river sediments. Can J Microbiol 45:520–529

    Article  CAS  Google Scholar 

  • Narro ML, Cerniglia CE, Van Baalen C, Gibson DT (1992) Metabolism of phenanthrene by the marine cyanobacterium Agmenellum quadruplicatum PR-6. Appl Environ Microbiol 58:1351–1359

    CAS  Google Scholar 

  • Ravindra K, Sokhi R, Van Grieken R (2008) Review: atmospheric polycyclic aromatic hydrocarbons: source attribution, emission factors and regulation. Atmos Environ 42:2895–2921

    Article  CAS  Google Scholar 

  • Sarma PM, Duraja P, Deshpande S, Lal B (2009) Degradation of pyrene by an enteric bacterium, Leclercia adecarboxylata PS4040. Biodegradation 21:59–69. doi:10.1007/s10532-009-9281-z

    Article  Google Scholar 

  • Schneider J, Grosser R, Jayasimhulu K, Xue W, Warshawsky D (1996) Degradation of pyrene, benz[a]anthracene and benzo[a]pyrene by Mycobacterium sp. strain RJGII-135, isolated from a former coal gasification site. Appl Environ Microbiol 62:13–19

    CAS  Google Scholar 

  • Vila J, Lopez Z, Sabate J, Minguillon C, Solanas AM, Grifoll M (2001) Identification of a novel metabolite in the degradation of pyrene by Mycobacterium sp. strain AP1: actions of the isolate on two and three-ring polycyclic aromatic hydrocarbons. Appl Environ Microbiol 67:5497–5505

    Article  CAS  Google Scholar 

  • Walter U, Beyer M, Klein J, Rehm HJ (1991) Degradation of pyrene by Rhodococcus sp. UW1. Appl Microbiol Biotechnol 34:671–676

    Article  CAS  Google Scholar 

  • Widada J, Nojiri H, Kasuga K, Yoshida T, Habe H, Omori T (2002) Molecular detection and diversity of polycyclic aromatic hydrocarbon degrading bacteria isolated from geographically diverse sites. Appl Microbiol Biotechnol 58:202–209

    Article  CAS  Google Scholar 

  • Wiele T, Vanhaecke VL, Boeckaert C, Peru K, Headley J, Verstraete W, Siciliano S (2005) Human colon microbiota transform polycyclic aromatic hydrocarbons to estrogenic metabolites. Environ Health Perspect 113:6–10

    Article  Google Scholar 

  • Wong JWC, Lai KM, Wan CK, Ma KK, Fang M (2002) Isolation and optimization of PAH-degradation bacteria from contaminated soil for PAHs bioremediation. Water Air Soil Pollut 139:1–13

    Article  CAS  Google Scholar 

  • Wunder T, Marr J, Kremer S, Sterner O, Anke H (1997) 1- Methoxypyrene and 1, 6-methoxypyrene: two novel metabolites in fungal metabolism of polycyclic aromatic hydrocarbons. Arch Microbiol 167:310–331

    Article  CAS  Google Scholar 

  • Yanna L, Gardner DR, Chen D, Anderson AJ, Weimer BC, Sims RC (2006) Study of biochemical pathways and enzymes involved in pyrene degradation by Mycobacterium sp. Strain KMS. Appl Environ Microbiol 72:7821–7828

    Article  Google Scholar 

  • Zaghden H, Kallel M, Elleuch B, Oudot J, Saliot A (2007) Sources and distribution of aliphatic and polyaromatic hydrocarbons in sediments of Sfax, Tunisia, Mediterranean Sea. Mar Chem 105:70–89

    Article  CAS  Google Scholar 

Download references

Acknowledgments

The authors are thankful to Director IITR, Lucknow for providing necessary facilities to do work. The authors are also thankful to Mr. Ashutosh Pathak for initial enrichment studies, Dr. Nasreen Gazi Ansari, Mr. Abhay and Prashant for help in the manuscript preparation.

Author information

Authors and Affiliations

  1. Analytical Chemistry Section, Indian Institute of Toxicological Research, CSIR, Lucknow, India

    Jitendra Nath Tiwari, Mudiam Mohana Krishna Reddy, Devendra Kumar Patel & Ramesh Chandra Murthy

  2. Environmental Biotechnology Division, Indian Institute of Toxicological Research, CSIR, Lucknow, India

    Natesan Manickam

  3. School of Studies in Microbiology, Vikram University, Ujjain, India

    Sudhir Kumar Jain

Authors
  1. Jitendra Nath Tiwari
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Mudiam Mohana Krishna Reddy
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Devendra Kumar Patel
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Sudhir Kumar Jain
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Ramesh Chandra Murthy
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Natesan Manickam
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Devendra Kumar Patel.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Tiwari, J.N., Reddy, M.M.K., Patel, D.K. et al. Isolation of pyrene degrading Achromobacter xylooxidans and characterization of metabolic product. World J Microbiol Biotechnol 26, 1727–1733 (2010). https://doi.org/10.1007/s11274-010-0350-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11274-010-0350-6

Keywords

Search

Navigation