Phialophora Melinii (NFCCI 3617): A Newly Isolated Psychrotolerant Fungus That Produces Enhanced Laccase Under the Influence of Organic Solvents

Kusum Dhakar, Pandey Anita


A psychrotolerant fungus, isolated from decomposing pine needle debris, is investigated for laccase production under the influence of 5 organic solvents. The fungus was identified as Phialophora melinii and was able to grow between 4 to 35 °C (opt. 25 °C) and 2-14 pH (opt. 5-7). In quantitative estimations that were carried out at optimum growth temperature and pH, the fungal laccase was estimated to be 21.0 ± 4.0 U/L. Native PAGE study revealed 35 kDa molecular mass of the fungal laccase. Supplementation of organic solvents namely, methanol, ethanol, acetone, n-propanol and iso-propanol in varying concentrations (0.5%-2.0%, separately), significantly affected the production of fungal laccase. Out of 5 solvents used, n-propanol was found to be the most efficient enhancer of laccase production. n-Propanol (0.5%) resulted in maximum enhancement (7 folds) in laccase production at 18th day of incubation. Methanol, iso-propanol and ethanol were able to enhance laccase production up to 5-6 folds in comparison to control with respect to the varying concentration and incubation length. Age of the fungal culture (incubation days) was observed as an important factor for laccase production. Use of low molecular compounds in enhancing the fungal laccase production may be considered as an eco-friendly approach.


Phialophora; Biodegradation; Laccase; Organic Solvents; Indian Himalayan region

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Alves, A. M. C. R., Record, E., Lomascolo, A., Scholtmeijer, K., Asther, M., Wessels, J. G. H., & Wosten, H. A. B. (2004). Highly efficient production of laccase by the basidiomycete Pycnoporus cinnabarinus. Applied and Environmental Microbiology, 70, 6379-6384.

Bertrand, B., Martinez-Morales, F., & Trejo-Hernandez, M. R. (2013). Fungal laccases: Induction and production. Revista Mexicana de Ingenieria Quimica, 12, 473-488.

Bettin, F., Montanari, Q., Calloni, R., Gaio, T. A., Silveira, M. M., & Dillon, A. J. P. (2014). Additive effects of CuSO4 and aromatic compouds on laccase production by Pleurotus sajor-caju PS 2001 using sucrose as a carbon source. Brazilian Journal of Chemical Engineering, 31, 335-346.

Bisht, D., Pandey, A., & Palni, L. M. S. (2003). Influence of microbial inoculations on Cedrus deodara in relation to survival, growth promotion and nutrient uptake of seedlings and general soil microflora. Journal of Sustainable Forestry, 17(3), 37-54.

Crowe, J. D., & Olsson, S. (2001). Induction of laccase activity in Rhizoctonia solani by antagonistic Pseuodomonas fluorescens strains and a range of chemical treatments. Applied and Environmental Microbiology, 67, 2088-2094.

Dhakar, K., Jain, R., Tamta, S., & Pandey, A. (2014a). Prolonged laccase production by a cold and pH tolerant strain of Penicillium pinophilum (MCC1049) isolated from a low temperature environment. Enzyme Research

Dhakar, K., & Pandey, A. (2013). Laccase production from a temperature and pH tolerant fungal strain of Trametes hirsuta (MTCC 11397). Enzyme Research. doi:org/10.1155/2013/86906

Dhakar, K., Sharma, A., & Pandey, A. (2014b). Cold, pH and salt tolerant Penicillium spp. inhabit the high altitude soils in Himalaya, India. World Journal of Microbiology and Biotechnology, 30, 1315-1324.

Dhawan, S., & Kuhad, R. C. (2002). Effect of amino acids and vitamins on laccase production by the bird’s nest fungus Cyathus bulleri. Bioresource Technology, 84, 35-38.

Dighton, J., White, J. F., & Oudemans, P. (2005). The fungal community: Its organization and role in the ecosystem. CRC Press (Taylor & Francis Group).

Elisashvili. V., Kachlishvili, E., Khardziani, T., & Agathos, S. N. (2010). Effect of aromatic compounds on the production of laccase and manganese peroxidase by white-rot basidiomycetes. Journal of Industrial Microbiology and Biotechnology, 37, 1091-1096.

Han, M. J., Choi, H. T., & Song, H. G. (2005). Purification and characterization of laccase from the white rot fungus Trametes versicolor. Journal of Microbiology, 43, 555-560.

Jeon, J. R., Baldrian, P., Murugesan, K., & Chang, Y. S. (2012). Laccase-catalysed oxidations of naturally occurring phenols: From in vivo biosynthetic pathways to green synthetic applications. Microbial Biotechnology, 5, 318-332.

Kirk, T. K., & Farrell, R. L. (1987). Enzymatic combustion- the microbial-degradation of lignin. Annual Review of Microbiology, 41, 465-505.

Kunamneni, A., Ballesteros, A., Plou, F. J., & Alcalde, M. (2007). Fungal laccase—A versatile enzyme for biotechnological applications. In A. Mendez-Vilas (Ed.), Communicating current research and educational topics and trends in applied microbiology (pp. 233-245). Formex, Badajoz, Spain.

Laemmli, U. K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227, 680-685.

Lee, I.Y ., Jung, K. H., Lee, C. H., & Park, Y. H. (1999). Enhanced production of laccase in Trametes vesicolor by the addition of ethanol. Biotechnology Letters, 21, 965-968.

Lomascolo, A., Record, E., Herpoel-Gimbert, I., Delattre, M., Robert, J. L., Georis, J., ... Asther, M. (2003). Over-production of laccase by a monokaryotic strain of Pycnoporus cinnabarinus using ethanol as inducer. Journal of Applied Microbiology, 94, 618-624.

Lu, L., Zeng, G., Fan, C., Ren, X., Wang, C., Zhao, Q., ... Jiang, M. (2013). Characterization of a laccase-like multicopper oxidase from newly isolated Streptomyces sp. C1 in agricultural waste compost and enzymatic decolorization of azo dyes. Biochemical Engineering Journal, 72, 70-76.

Mishra, P. K., Bisht, S. C., Bisht, J. K., & Bhatt, J. C. (2012). Cold tolerant PGPRs as bioinoculants for stress management. In D.K. Maheshwari (Ed.), Bacteria in Agrobiology: Stress Management (pp.95-118). Springer, Berlin, Germany.

Novotny, C., Svobodova, K., Erbanova, P., Cajthamla, T., Kasinatha, A., Lang, E., & Sasek, V. (2004). Ligninolytic fungi in bioremediation: Extracellular enzyme production and degradation rate. Soil Biology and Biochemistry, 36, 1545-1151.

Pandey, A., & Palni, L.M.S. (2007). The rhizosphere effect in trees of the Indian central Himalaya with special reference to altitude. Applied Ecology and Environmental Research, 5(1), 93-102.

Pandey, A., Palni, L .M. S., & Bisht, D. (2001a). Dominant fungi in the rhizosphere of established tea bushes and their interactions with the dominant bacteria under in situ conditions. Microbiological Research, 156, 377-382.

Pandey, A., Palni, L. M. S., & Hebbar, K. P. (2001b). Suppression of damping-off in maize seedlings by Pseudomonas corrugata. Microbiological Research, 156, 191-194.

Rinu, K., & Pandey, A. (2010). Temperature dependent phosphate solubilization by cold and pH tolerant species of Aspergillus isolated from Himalayan soil. Mycoscience, 51, 263-271.

Rinu, K., & Pandey, A. (2011). Slow and steady phosphate solubilization by a psychrotolerant strain of Paecilomyces hepiali (MTCC 9621). World Journal of Microbiology and Biotechnology, 27, 1055-1062.

Rinu, K., Pandey, A., & Palni, L. M. S. (2012). Utilization of psychrotolerant phosphate solubilizing fungi under low temperature conditions of the mountain ecosystem. In T. Satyanarayana, B. N. Johri & A. Prakash (Eds.), Microorganisms in Sustainable Agriculture and Biotechnology (pp.77-90). Springer Science + Buisiness Media.

Rivera-Hoyos, C. M., Morales-Alvarez, E. D., Poutou Pinales, R. A., Pedroza-Rodriguez, A. M., Rodriguez-Vazquez, R., & Delgado-Boada, J. M. (2013). Fungal laccases. Fungal Biology Reviews, 27, 67-82.

Rizvi, S. J., & Rizvi, S. J. (1992). Allelopathy-basic and applied aspects. Chapman and Hall, London.

Rodgers, C. J., Blanford, C. F., Giddens, S. R., Skamnioti, P., Armstrong, F. A., & Gurr, S. J. (2010). Designer laccases: A vogue for high-potential fungal enzymes? Trends in Biotechnology, 28, 63-72.

Sati, P., Dhakar, K., & Pandey, A. (2013). Microbial diversity in soil under potato cultivation from cold desert Himalaya, India. ISRN Biodiversity. doi: org/10.1155/2013/767453

Singh, S. M., Singh, S. K., Yadav, L. S., Singh, P. N., & Ravindra, R. (2012). Filamentous soil fungi from Ny-Ålesund, Spitsbergen, and screening for extracellular enzymes. Arctic, 65, 45-55.

Strong, P. J., & Claus, H. (2011). Laccase: A review of its past and its future in bioremediation. Critical Reviews in Environmental Science and Technology, 4, 373-434.

Trivedi, P., Kumar, B., Pandey, A., & Palni, L. M. S. (2007). Growth promotion of rice by phosphate solubilizing bioinoculants in a Himalayan location. In E. Velazqez & C. Rodriguez-Barrueco (Eds.), Plant and soil, developments in plant and soil sciences (Vol.102, pp.291-299). First International Meeting on Microbial Phosphate Solubilization, Springer.

Trivedi, P., & Pandey, A. (2008). Plant growth promotion abilities and formulation of Bacillus megaterium strain B 388 (MTCC6521) isolated from a temperate Himalayan location. Indian Journal of Microbiology, 48(3), 342-


Yarzábal, L. A. (2014). Cold-tolerant phosphate-solubilizing microorganisms and agriculture development in mountainous regions of the world. In M. S. Khan et al. (Eds.), Phosphate Solubilizing Microorganisms. doi: 10.1007/978-3-319-08216-5_5, © Springer International Publishing Switzerland.




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