Lignocellulolytic activities of a novel strain of Trichoderma harzianum

M.  Rubeena; Kannan  Neethu; S.  Sajith; S.  Sreedevi; Prakasan  Priji; K. N.  Unni; M. K. Sarath  Josh; V. N.  Jisha; S.  Pradeep; Sailas  Benjamin

doi:10.4236/abb.2013.42030

M. Rubeena, Kannan Neethu, S. Sajith, S. Sreedevi, Prakasan Priji, K. N. Unni, M. K. Sarath Josh, V. N. Jisha, S. Pradeep, Sailas Benjamin

Abstract: This study describes a novel dark-green spore producing strain of Trichoderma harzianum exhibiting higher activities of cellulase, hemicellulase and ligninase on specific plate assays. To assess the cellulase production in detail, basal salt medium (BSM) was supplemented with synthetic [carboxymethyl cellulose (CMC), glucose, sucrose, dextrose, lactose or maltose] and natural (flours of banana, banana peel, jack seed, potato or tapioca) carbon as well as nitrogen (yeast extract, beef extract, peptone, NaNO3 or NH4NO3) sources. Temperature and pH optima were 28?C and 4, respectively for the growth of the fungus in CMC- BSM with 146 U/ml cellulase activity. Flours of potato and banana supported comparable yields of cellulase to that of CMC (147 U/ml and 168 U/ml, respectively), while sodium nitrate was the preferred nitrogen source (150 U/ml). The water soluble yellowish-green pigment (a probable siderophore) extracted from the spores showed an absorption maximum at 414 nm. To comprise, this fungus shows the complete lignocellulolytic potential which offers great industrial significance, especially for the ethanol production from the lignocellulosic waste coupled with the production of a new pigment.

Keywords: Trichoderma harzianum; Morphology; Submerged Fermentation; Yellowish-Green Pigment

Cite this paper: Rubeena, M. , Neethu, K. , Sajith, S. , Sreedevi, S. , Priji, P. , Unni, K. , Josh, M. , Jisha, V. , Pradeep, S. and Benjamin, S. (2013) Lignocellulolytic activities of a novel strain of Trichoderma harzianum. Advances in Bioscience and Biotechnology, 4, 214-221. doi: 10.4236/abb.2013.42030.

References

[1] Behrendt, C.J., Blanchette, R.A., Akhtar, M., Enebak, S., Iverson, S. and Williams, D. (2000) Biomechanical pulping with Phlebiopsis gigantea reduced energy consumption and increased paper strength. Tappi Journal, 83, 65.

[2] Blanchette, R.A., Behrendt, C.D., Williams, D., Iverson, S., Akhtar, M. and Enebak, S.A. (1998) A new approach to effective biopulping: Treating logs with Phlebiopsis gigantean. 7th International Conference on Biotechnology in the Pulp and Paper Industry, A51-A54.

[3] Ruiz-Duenas, F.J. and Martínez, A.T. (2009) Microbial degradation of lignin: How a bulky recalcitrant polymer is efficiently recycled in nature and how we can take advantage of this. Microbial Biotechnology, 2, 164-177. doi:10.1111/j.1751-7915.2008.00078.x

[4] Balakrishnan, H., Choudhury, M.D., Srinivasan, M.C. and Rele, M.V. (1992) Cellulase-free xylanase production from an alkalophilic Bacillus species. World Journal of Microbioliology Biotechnology, 8, 627-631. doi:10.1007/BF01238802

[5] Viikari, L., Kantelinen, A., Sundquist, J. and Linko, M. (1994) Xylanases in bleaching: from an idea to the industry. FEMS Microbiology Reviews, 13, 335-350. doi:10.1111/j.1574-6976.1994.tb00053.x

[6] Singhania, R.R., Sukumaran, R.K. and Pillai, A. (2006) Solid-state fermentation of lignocellulosic substrates for cellulase production by Trichoderma reesei NRRL 11460. Indian journal of Biotechnology, 5, 332-336.

[7] Neethu, K., Rubeena, M., Sajith, S., Sreedevi, S., Priji, P., Unni, K.N., Sarath Josh, M.K., Jisha, V.N., Pradeep, S. and Benjamin, S. (2012) Advances in Bioscience and Biotechnology, 3, 1160-1166. doi:10.4236/abb.2012.38142

[8] Tengerdy, R.P. (1996) Cellulase production by solid substrate fermentation. Indian Journal of Scientific Research, 55, 313-316.

[9] Viterbo, A., Haran, S., Friesem, D., Ramot, O. and Chet, I. (2001) Antifungal activity of a novel endochitinase gene (chit36) from Trichoderma harzianum Rifai TM. FEMS Microbiology Letters, 200, 169-174. doi:10.1111/j.1574-6968.2001.tb10710.x

[10] Castro, A.M., Pedro, K.C., Cruz, J.C., Ferreira, M.C., Leite, S.G. and Pereira, N. (2010) Trichoderma harzianum IOC-4038: A promising strain for the production of a cellulolytic complex with significant β-glucosidase activity from sugarcane bagasse cellulignin. Applied Biochemistry and Biotechnology, 162, 2111-2122. doi:10.1007/s12010-010-8986-0

[11] Colussi, F., Garcia, W., Rosseto, F.R., Mello, B.L.S., Net, M.O. and Polikarpov, I. (2011) Effect of pH and temperature on the global compactness, structure, and activity of cellobiohydrolase Cel7A from Trichoderma harzianum. Journal of European Biophysics, 40, 89-98.

[12] Stockton, B.C., Mitchell, D.J. and Grohmann K. (1991) Optimum β-D glucosidase supplementation of cellulose for efficient conversion of cellulose to glucose. Biotechnology Letters, 13, 57-62. doi:10.1007/BF01033518

[13] Berry, D.R. and Paterson, A. (1990) Enzymes in food industry. In: Suckling, C.J., Ed., Enzyme Chemistry, Impact and Applications, 306-351.

[14] Miller, G.L. (1959) Use of dinitrosalicyclic acid reagent for determination of reducing sugar. Analytical Chemistry, 31, 426-428. doi:10.1021/ac60147a030

[15] Sazci, A., Erenler, K. and Radford, A. (1986) Detection of cellulolytic fungi by using Congo red as an indicator: A comparative study with the dinitrosalicyclic acid reagent method. Journal of Applied Microbiology, 61, 559562. doi:10.1111/j.1365-2672.1986.tb01729.x

[16] Kuhad, R.C., Singh, A. and Eriksson, K.E.L. (1997) Microorganisms and enzymes involved in the degradation of plant fiber cell walls. Advances in Biochemical Engineering/Biotechnology, 57, 45-125. doi:10.1007/BFb0102072

[17] Gashe, B.A. (1992) Cellulase production and activity by Trichoderma sp. A-001. Journal of Applied Microbiology, 73, 79-82. doi:10.1111/j.1365-2672.1992.tb04973.x

[18] Mekala, N.K., Singhania, R.R., Sukumaran, R.K. and Pandey (2008) Cellulose production under solid-state fermentation by Trichoderma ressei RUT C30: Statistical optimization of process parameters. Applied Biochemistry and Biotechnology, 151, 122-131. doi:10.1007/s12010-008-8156-9

[19] Juhasz, T.Z., Szengyel, N., Szijarto and Reczey, K. (2004) Effect of pH on cellulases production of Trichoderma reesei RUT C30. Applied Biochemistry and Biotechnology, 201, 113-116.

[20] Deschamps, F., Giuliano, C., Asther, M., Huet, M.C and Roussost, S. (1985) Cellulase Production by Trichoderma harzianum in static and mixed solid-state fermentation reactors under non aseptic condition. Biotechnology and Bioengineering, 27, 1385-1388. doi:10.1002/bit.260270917

[21] Niranjane, A.P., Madhou, P. and Stevenson, T.W. (2006) The effect of carbohydrate carbon sources on the production of cellulase by Phlebia gigantean. Enzyme and Microbial Technology, 40, 1464-1468. doi:10.1016/j.enzmictec.2006.10.041

[22] Xia, L. and Cen, P. (1999) Cellulase production by solid state fermentation on lignocellulosic waste from the xylose industry. Process Biochemistry, 34, 909-912. doi:10.1016/S0032-9592(99)00015-1

[23] Wen, Z., Liao, W. and Chen, S. (2005) Production of cellulase/β-glucosidase by the mixed fungi culture Trichoderma reesei and Aspergillus phoenicis on dairy manure. Process Biochemistry, 40, 3087-3094. doi:10.1016/j.procbio.2005.03.044

[24] Camassola, M. and Dillon, A.J.P. (2007) Production of cellulases and hemicellulases by Penicillium echinulatum grown on pretreated sugar cane bagasse and wheat bran in solid-state fermentation. Journal of Applied Microbiology, 103, 2196-2204. doi:10.1111/j.1365-2672.2007.03458.x

[25] Kiss, G.C., Forgacs, E., Cserhati, T. and Vizcaino, J.A. (2000) Colour pigments of T. harzianum preliminary investigations with thin-layer chromatography—Fourier transform infrared spectroscopy and high-performance liquid chromatography with diode array and mass spectrometric detection. Journal of Chromatography A, 896, 61-68.

[26] Sharma, D., Gupta, C., Aggarwal, S. and Nagpal, N. (2012) Pigment extraction from fungus for textile dyeing. Indian Journal of Fibre and Textile Research, 37, 68-73.

[27] Betina, V. (1995) Photoinduced conidiation in Trichoderma viride. Folia Microbiologica, 40, 219-224. doi:10.1007/BF02814196

[28] Xiaoyi1, Z., Yingde, C.U., Ning, L.U., Xin, Y.U. and Meimei, Z. (2012) Optimization of fermentation condition in liquid culture of T. viride to produce yellow pigment. Center for Improved Engineering and Science Education, 61, 3205-3212.

[29] Chitale, A., Jadhav, D.V., Waghmare, S.R., Sahoo, A.K. and Ranveer, R.C. (2012) Production and characterization of brown coloured pigment from Trichoderma viride. Electronic Journal of Environmental Agricultural and Food Chemistry, 11, 529-537.