doi: 10.15389/agrobiology.2019.1.65eng

UDC 579.64:579.222.2

We are grateful to Prof. G.C. Varese (Department of Life Sciences and Systems Biology, University of Turin) for kindly providing us with fungal strains.

Supported financially by Russian Science Foundation (grant No. 16-14-00081, the PAH degradation part), and by Russian Foundation for Basic Research (grant No. 18-29-05062, the oil degradation part)



O.V. Turkovskaya, E.V. Dubrovskaya, V.S. Grinev, S.A. Balandina,
N.N. Pozdnyakova

Institute of Biochemistry and Physiology of Plants and Microorganisms RAS, 13, prosp. Entuziastov, Saratov, 410049 Russia, e-mail (✉ corresponding author),,,,

Turkovskaya O.V.
Balandina S.A.
Dubrovskaya E.V.
Pozdnyakova N.N.
Grinev V.S.

Received August 15, 2018


Environmental pollution by natural and man-made pollutants remains a serious problem. Agricultural areas are contaminated by major and hazardous pollutants such as oil, which comes from local oil-producing and oil-refining facilities, and polycyclic aromatic hydrocarbons (PAHs), which result from natural fires and from human activity associated with the use of flammable organic raw materials. This presents the hazard of accumulation of toxic substances in food and fodder plants. Natural ecosystems have powerful detoxifying potential, which is ensured by the degradative activity of microorganisms, including ascomycetes — one of the largest groups in the fungal kingdom. Here we examined the degradation of oil and PAHs by micromycetes with different ecological strategies and detected ligninolytic enzymes implicated in the oxidation of the pollutants. We used four ascomycete strains with different taxonomic affiliations and ecological strategies. These were Fusarium oxysporum IBPPM543, Lecanicillium aphanocladii IBPPM542, Cladosporium herbarum MUT3238, and Geotrichum candidum MUT4803. The fungi were grown in liquid media with different compositions that received additions of the pollutants used: oil, PAHs, and anthraquinone-type dyes. After 14 days of fungal growth, the elimination of the pollutants and the content of their main degradation products were examined by GC. Ligninolytic enzyme activity was estimated spectrophotometrically by the oxidation rate of the corresponding test substrates. All treatments in the experiments and analyses had no less than three replications, and each experiment was repeated no less than three times. Data were processed with Microsoft Excel 2003 software. All fungi oxidized oil; the utilization was from 46 to 82 % of the initial concentration of 5 g/l within 14 days. C. herbarum MUT 3238 metabolized all PAHs included in the study (anthracene, phenanthrene, and fluorene) almost completely (initial concentration, 0.05 g/l). L. aphanocladii IBPPM 542 degraded anthracene, phenanthrene, and fluorene by 40, 63, and 81 %, respectively. F. oxysporum IBPPM 543 utilized phenanthrene and fluorene only by 20 and 40 %, respectively. PAH degradation by G. candidum MUT4803 was not greater than 18 %. Anthracene was not degraded by F. oxysporum IBPPM 543 and G. candidum MUT4803. The degradation of the pollutants was accompanied by the production of extracellular peroxidases by all fungi except G. candidum. The activities of these peroxidases were largely stimulated by Mn2+; this property makes them similar to the Mn-peroxidases of basidiomycetes. This is the first report on the production of extracellular peroxidases by C. herbarum and L. aphanocladii. Neither of the fungi produced lignin peroxidase or laccase. Identification of the PAH oxidation products allowed us to suggest a pathway for PAH degradation by the tested fungi with an extracellular Mn-peroxidase. The degradation proceeds through the formation of quinones and carboxylic acids (phthalic and 2,2'-diphenic), which indicates that the PAHs are utilized almost completely and that no toxic metabolites accumulate. The obtained results indicate that two widely distributed ascomycete species, C. herbarum and F. oxysporum, and a strain of the lesser-known and poorly studied species L. aphanocladii, have degradative potential toward oil and PAHs, which presupposes their involvement in the self-cleaning of the environment from these pollutants. The detection of ligninolytic enzymes (Mn-peroxidases) and of the corresponding products of PAH degradation speaks in favor of an ecologically appropriate pathway for the utilization of PAHs, which reduces the negative consequences associated with the possible formation of toxic metabolites. In the G. candidum strain, the oxidation of oil and PAHs is possibly due to the activity of other enzymes, for example cytochrome Р450 monooxygenase, because no ligninolytic enzymes have been found. In addition, it is highly possible that this strain has a “dye peroxidase”, which requires a narrow range of substrates and catalyzes the degradation of anthraquinone dyes, as was also shown by us. The ability of all fungal strains to degrade pollutants makes them promising candidates for practical use in bioremediation and other biotechnologies.

Keywords: ascomycetes, Fusarium oxysporum, Lecanicillium aphanocladii, Cladosporium herbarum, biodegradation, polycyclic aromatic hydrocarbons, oil, ligninolytic enzymes, peroxidases.




  1. Vasil'ev A.V., Bykov D.E., Pimenov A.A. Izvestiya Samarskogo nauchnogo tsentra RAN, 2015, 17(4): 269-272 (in Russ.)
  2. Gorobtsova O.N., Nazarenko O.G., Minkina T.M., Borisenko N.I., YAroshchuk A.V. Izvestiya vuzov. Severo-Kavkazskii region. Estestvennye nauki, 2005, 1: 73-78 (in Russ.)
  3. Gagkaeva T.Yu., Shamshev I.V., Gavrilova O.P., Selitskaya O.G. Biological relationships between Fusarium fungi and insects (review).Sel’skokhozyaistvennaya Biologiya [Agricultural Biology], 2014, 3: 13-23 CrossRef (in Russ.)
  4. Dinolfo M.I., Castañares E., Stenglei S.A. Fusariumplant interaction: state of the art a review. Plant Protect. Sci., 2017, 53: 61-70 CrossRef
  5. Gordon T.R., Okamoto D., Jacobson D.J. Colonization of muskmelon and nonsusceptible crops by Fusarium oxysporum f. sp. melonis and other species of Fusarium. Phytopathology, 1989, 79(10): 1095-1100 CrossRef
  6. Lemanceau P., Bakker P.A.H.M., DeKogel W.J., Alabouvette C., Schippers B. Antagonistic effect of nonpathogenic Fusarium oxysporum Fo47 and pseudobactin 358 upon pathogen Fusarium oxysporum f. sp. dianthi. Appl. Environ. Microbiol., 1993, 59(1): 74-82.
  7. Jacques R.J., Okeke B.C., Bento F.M., Teixeira A.S., Peralba M.C., Camargo F.A. Microbial consortium bio ugmentation of a polycyclic aromatic hydrocarbons contaminated soil. Bioresource Technol., 2008, 99(7): 2637-2643 CrossRef
  8. Thion C., Cébron A., Beguiristain T., Leyval C. Inoculation of PAH-degrading strains of Fusarium solani and Arthrobacter oxydans in rhizospheric sand and soil microcosms: microbial interactions and PAH dissipation. Biodegradation, 2013, 24(4): 569-581 CrossRef
  9. Krivobok S., Miriouchkine E., Seigle-Murandi F., Benoit-Guyod J.-L. Biodegradation of anthracene by soil fungi. Chemosphere, 1998, 37(4): 523-530 CrossRef
  10. Potin O., Veignie E., Rafin C. Biodegradation of polycyclic aromatic hydrocarbons (PAHs) by Cladosporium sphaerospermum isolated from an aged PAH contaminated soil. FEMS Microbiol. Ecol., 2004, 51(1): 71-78 CrossRef
  11. Boutrou R., Guéguen T.M. Interests in Geotrichum candidum for cheese technology. Int. J. Food Microbiol., 2005, 102(1): 1-20 CrossRef
  12. Kim S.J., Ishikawa K., Hirai M., Shoda M. Characteristics of a newly isolated fungus, Geotrichum candidum Des 1, which decolorizes various dyes. Journal of Fermentation and Bioengineering, 1995, 79(6): 601-607 CrossRef
  13. Kim S.J., Shoda M. Purification and characterization of a novel peroxidase from Geotrichum candidum Dec 1 involved in decolorization of dyes. Appl. Environ. Microb., 1999, 65(3): 1029-1035.
  14. Ziganshin A.M., Gerlach R., Naumenko E.A., Naumova R.P. Aerobic degradation of 2,4,6-trinitrotoluene by the yeast strain Geotrichum candidum AN-Z4. Microbiology, 2010, 79(2): 178-183 CrossRef
  15. Jakovljević V.D., Vrvić M.M. Potential of pure and mixed cultures of Cladosporium cladosporioides and Geotrichum candidum for application in bioremediation and detergent industry. Saudi J. Biol. Sci., 2018, 25(3): 529-536 CrossRef
  16. Zare R., Gams W. A revision of Verticillium section Prostrata. IV. The genera Lecanicillium and Simplicillium gen. nov. Nova Hedwigia, 2001, 73(1/2): 1-50.
  17. Manfrino R.G., González A., Barneche J., Tornesello Galván J., Hywell-Jones N., López- Lastra C.C. Contribution to the knowledge of pathogenic fungi of spiders in Argentina. Southernmost record in the world. Rev. Argent. Microbiol., 2017, 49(2): 197-200 CrossRef
  18. El-Debaiky S.A. New record of Lecanicillium aphanocladii family: Cordycipitaceae from Egypt. J. Bacteriol. Mycol. Open Access, 2017, 5(7): 00161 CrossRef
  19. Pinto A., Serrano C., Pires T., Mestrinho E., Dias L., Teixeira D., Caldeira A. Degradation of terbuthylazine, difenoconazole and pendimethalin pesticides by selected fungi cultures. Sci. Total Environ., 2012, 435-436(1): 402-410 CrossRef
  20. Vroumsia T., Steiman R., Seigle-Murandi F., Benoit-Guyod J.-L. Effects of culture parameters on the degradation of 2,4-dichlorophenoxyacetic acid (2,4-D) and 2,4-dichlorophenol (2,4-DCP) by selected fungi. Chemosphere, 1999, 39(9): 1397-1405 CrossRef
  21. Kadri T., Rouissi T., Brar S.K., Cledon M., Sarma S., Verma M. Biodegradation of polycyclic aromatic hydrocarbons (PAHs) by fungal enzymes: A review. J. Environ. Sci., 2017, 51(1): 52-74 CrossRef
  22. Ghosal D., Ghosh S., Dutta T.K., Ahn Y. Current state of knowledge in microbial degradation of polycyclic aromatic hydrocarbons (PAHs): A Review. Front. Microbiol., 2016, 7: 1369 CrossRef
  23. Bezalel L., Hadar Y., Cerniglia C. Enzymatic mechanisms involved in phenanthrene degradation by the white rot fungus Pleurotus ostreatus. Appl. Environ. Microbiol., 1997, 63(7): 2495-2501.
  24. Kirk T., Croan S., Tien M., Murtagh K., Farrell R. Production of multiple ligninases by Phanerochaete chrysosporium effect of selected growth condition and use mutant strain. Enzyme Microb. Tech., 1986, 8(1): 27-32.
  25. Pozdnyakova N.N., Jarosz-Wilkolazka A., Polak J., Graz M., Turkovskaya O.V. Decolourisation of anthraquinone-and anthracene-type dyes by versatile peroxidases from Bjerkandera fumosa and Pleurotus ostreatus D1. Biocatal. Biotransform., 2015, 33(2): 69-80 CrossRef
  26. Metody analiza organicheskogo veshchestva porod, nefti i gaza /Pod redaktsiei A.V. Ryl'kova [Methods for analyzing rock organic matter, oil and gas. A.V. Ryl'kov (ed.)]. Tyumen', 1977 (in Russ.).
  27. Niku-Paavola M.L., Karhunen E., Salola P., Raunio V. Ligninolytic enzymes of the white rot fungus Phlebia radiata. Biochem. J., 1988, 254(3): 877-884 CrossRef
  28. Heinfling A., Martinez M., Martinez A., Bergbauer M., Szewzyk U. Purification and characterization of peroxidases from dye-decolorizing fungus Bjerkandera adusta. FEMS Microbiol. Lett., 1998, 165(1): 43-50 CrossRef
  29. Tien M., Kirk K. Lignin-degrading enzyme from Phanerochaete chrysosporium: purification, characterization, and catalytic properties of a unique H2O2-requiring oxygenase. PNAS USA, 1984, 81(8): 2280-2284 CrossRef
  30. Sardrood B.P., Goltapeh E.M., Varma A. An introduction to bioremediation. In: Fungi as Bioremediators. Soil Biology, V. 32. E. Goltapeh, Y. Danesh, A. Varma (eds.). Springer, Berlin, Heidelberg, 2013 CrossRef
  31. Mohsenzadeh F., Nasseri S., Mesdaghinia A., Nabizadeh R., Zafari D., Khodakaramian G., Chehregani A. Phytoremediation of petroleum-polluted soils: Application of Polygonum aviculare and its root-associated (penetrated) fungal strains for bioremediation of petroleum-polluted soils. Ecotox. Environ. Safe., 2010, 73(4): 613-619 CrossRef
  32. Varjani S.J. Microbial degradation of petroleum hydrocarbons. Bioresource Technol., 2017, 223(1): 277-286 CrossRef
  33. Balaji V., Arulazhagan P., Ebenezer P. Enzymatic bioremediation of polyaromatic hydrocarbons by fungal consortia enriched from petroleum contaminated soil and oil seeds. J. Environ. Biol., 2014, 35: 521-529.
  34. Wong D.W.S. Structure and action mechanism of ligninolytic enzymes. Appl. Biochem. Biotech., 2009, 157(2): 174-209 CrossRef
  35. Obruca S., Marova I., Matouskova P., Haronikova A., Lichnova A. Production of lignocellulose-degrading enzymes employing Fusarium solani F-552. Folia Microbiol., 2012, 57(3): 221-227 CrossRef
  36. Kwiatos N., Ryngaj??o M., Bielecki S. Diversity of laccase-coding genes in Fusarium oxysporum genomes. Front. Microbiol., 2015, 6: 33 CrossRef
  37. Wua Y.-R., Luo Z.-H., Vrijmoed L.L.P Biodegradation of anthracene and benz[a]anthracene by two Fusarium solani strains isolated from mangrove sediments. Bioresource Technol., 2010, 101(24): 9666-9672 CrossRef
  38. Sampedro I., D'Annibale A., Ocampo J.A., Stazi S.R., García-Romera I. Solid-state cultures of Fusarium oxysporum transform aromatic components of olive-mill dry residue and reduce its phytotoxicity. Bioresource Technol., 2007, 98(18): 3547-3554 CrossRef
  39. Asses N., Ayed L., Bouallagui H., Sayadi S., Hamdi M. Biodegradation of different molecular-mass polyphenols derived from olive mill wastewaters by Geotrichum candidum. International Biodeterioration & Biodegradation, 2009, 63(4): 407-413 CrossRef
  40. Ayed L., Assas N., Sayadi S., Hamdi M. Involvement of lignin peroxidase in the decolourization of black olive mill wastewaters by Geotrichum candidum. Lett. Appl. Microbiol., 2005, 40(1): 7-11 CrossRef
  41. Hammel K., Green B., Gai W. Ring fission of anthracene by eukaryote. PNAS USA, 1991, 88(23): 10605-10608 CrossRef
  42. Pozdnyakova N.N., Chernyshova M.P., Grinev V.S., Landesman E.O., Koroleva O.V., Turkovskaya O.V. Prikladnaya biokhimiya i mikrobiologiya, 2016, 52(6): 590-598 CrossRef (in Russ.).
  43. Pozdnyakova N., Dubrovskaya E., Chernyshova M., Makarov O., Golubev S., Balandina S., Turkovskaya O. The degradation of three-ringed polycyclic aromatic hydrocarbons by wood-inhabiting fungus Pleurotus ostreatus and soil-inhabiting fungus Agaricus bisporus. Fungal Biology, 2018, 122(5): 363-372 CrossRef
  44. Ning D., Wang H., Ding C., Lu H. Novel evidence of cytochrome P450-catalyzed oxidation of phenanthrene in Phanerochaete chrysosporium under ligninolytic conditions. Biodegradation, 2010, 21(6): 889-901 CrossRef






Full article PDF (Rus)

Full article PDF (Eng)