Applied Microbiology and Biotechnology

, Volume 91, Issue 4, pp 1227–1238 | Cite as

Isolation and characterization of four novel Gram-positive bacteria associated with the rhizosphere of two endemorelict plants capable of degrading a broad range of aromatic substrates

  • Lidija Djokic
  • Tanja Narancic
  • Jasmina Nikodinovic-Runic
  • Miloje Savic
  • Branka VasiljevicEmail author
Environmental Biotechnology


Four new Gram-positive, phenol-degrading strains were isolated from the rhizospheres of endemorelict plants Ramonda serbica and Ramonda nathaliae known to exude high amounts of phenolics in the soil. Isolates were designated Bacillus sp. PS1, Bacillus sp. PS11, Streptomyces sp. PS12, and Streptomyces sp. PN1 based on 16S rDNA sequence and biochemical analysis. In addition to their ability to tolerate and utilize high amounts of phenol of either up to 800 or up to 1,400 mg l−1 without apparent inhibition in growth, all four strains were also able to degrade a broad range of aromatic substrates including benzene, toluene, ethylbenzene, xylenes, styrene, halogenated benzenes, and naphthalene. Isolates were able to grow in pure culture and in defined mixed culture on phenol and on the mixture of BTEX (benzene, toluene, ethylbenzene, and xylenes) compounds as a sole source of carbon and energy. Pure culture of Bacillus sp. PS11 yielded 1.5-fold higher biomass amounts in comparison to mixed culture, under all conditions. Strains successfully degraded phenol in the soil model system (2 g kg−1) within 6 days. Activities of phenol hydroxylase, catechol 1,2-dioxygenase, and catechol 2,3-dioxygenase were detected and analyzed from the crude cell extract of the isolates. While all four strains use ortho degradation pathway, enzyme indicative of meta degradation pathway (catechol 2,3-dioxygenase) was also detected in Bacillus sp. PS11 and Streptomyces sp. PN1. Phenol degradation activities were induced 2 h after supplementation by phenol, but not by catechol. Catechol slightly inhibited activity of catechol 2,3-dioxygenase in strains PS11 and PN1.


Biodegradation Phenol BTEX Bacillus Streptomyces Soil model experiments 



This work was supported by the Ministry of Science and Technological Development of the Republic of Serbia (Grant no. 173048). The authors would like to thank Prof. Ljubiša Topisirović, Prof. Branka Stevanović, and Prof. Vladimir Stevanović for providing the samples, and also to Prof. Ananda Chakrabarty for useful comments on this work.

Supplementary material

253_2011_3426_MOESM1_ESM.pdf (193 kb)
Online Resource—Supplementary Fig. 1 Phylogenetic trees of 16S rRNA gene sequences based on neighbor-joining analyses showing the relationship of phenol-degrading bacteria and related species: a genus Bacillus, b genus Streptomyces. Isolated phenol degrading strains are designated in bold. GenBank accession numbers are in brackets. Bootstrap values were calculated by resampling method with 100 replicates and shown as percentage. Final plot was done according to Huson et al. (2007). (PDF 192 kb)


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Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Lidija Djokic
    • 1
  • Tanja Narancic
    • 1
  • Jasmina Nikodinovic-Runic
    • 1
  • Miloje Savic
    • 1
  • Branka Vasiljevic
    • 1
    Email author
  1. 1.Institute of Molecular Genetics and Genetic EngineeringUniversity of BelgradeBelgradeSerbia

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