Archives of Microbiology

, Volume 201, Issue 10, pp 1369–1383 | Cite as

Toluene degradation via a unique metabolic route in indigenous bacterial species

  • Fatima Muccee
  • Samina EjazEmail author
  • Naheed Riaz
Original Paper


Tanneries are the primary source of toluene pollution in the environment and toluene due to its hazardous effects has been categorized as persistent organic pollutant. Present study was initiated to trace out metabolic fingerprints of three toluene-degrading bacteria isolated from tannery effluents of Southern Punjab. Using selective enrichment and serial dilution methods followed by biochemical, molecular and antibiotic resistance analysis, isolated bacteria were subjected to metabolomics analysis. GC–MS/LC–MS analysis of bacterial metabolites helped to identify toluene transformation products and underlying pathways. Three toluene-metabolizing bacteria identified as Bacillus paralicheniformis strain KJ-16 (IUBT4 and IUBT24) and Brevibacillus agri strain NBRC 15538 (IUBT19) were found tolerant to toluene and capable of degrading toluene. Toluene-degrading potential of these isolates was detected to be IUBT4 (10.35 ± 0.084 mg/h), IUBT19 (14.07 ± 3.14 mg/h) and IUBT24 (11.1 ± 0.282 mg/h). Results of GC–MS analysis revealed that biotransformation of toluene is accomplished not only through known metabolic routes such as toluene 3-monooxygenase (T3MO), toluene 2-monooxygenase (T2MO), toluene 4-monooxygenase (T4MO), toluene methyl monooxygenase (TOL), toluene dioxygenase (Tod), meta- and ortho-ring fission pathways. But additionally, confirmed existence of a unique metabolic pathway that involved conversion of toluene into intermediates such as cyclohexene, cyclohexane, cyclohexanone and cyclohexanol. LC–MS analysis indicated the presence of fatty acid amides, stigmine, emmotin A and 2, 2-dinitropropanol in supernatants of bacterial cultures. As the isolated bacteria transformed toluene into relatively less toxic molecules and thus can be preferably exploited for the eco-friendly remediation of toluene.


Toluene-degrading bacteria Bioremediation Consortium Brevibacillus agri Bacillus paralicheniformis Burkholderia lata 



Volatile organic compound


Environmental Protection Agency


Permissible exposure limit


Dioxygenase-mediated pathway


Toluene-2-monooxygenase pathway


Toluene-3-monooxygenase pathway


Toluene-4-monooxygenase pathway


Toluene methyl monooxygenase pathway


Gas chromatography–mass spectrometry


Liquid chromatography–mass spectrometry


Basic local alignment search tool


Acetylcholinesterase inhibitor



We are thankful to Dr. Asma Shaukat, Department of Pathology, Quaid-e-Azam Medical College, Bahawalpur, for providing us the antibiotic sensitivity discs.

Author contributions

SE conceived the idea and designed study. FM performed all bench top work and wrote first draft of manuscript. NR interpreted the results of GC–MS analysis. All the authors contributed to finalize the manuscript.


This research did not receive any grant from funding agencies in the public, commercial or not-for-profit sectors.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflicts of interest.

Human and animal rights statement

This article does not contain any studies with human participants or animals performed by any of the authors.

Supplementary material

203_2019_1705_MOESM1_ESM.docx (688 kb)
Supplementary file1 (DOCX 687 kb)


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© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Biochemistry and BiotechnologyThe Islamia University of BahawalpurBahawalpurPakistan
  2. 2.Department of ChemistryThe Islamia University of BahawalpurBahawalpurPakistan

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