A novel strain of Trichoderma viride shows complete lignocellulolytic activities

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

Enzyme Technology Laboratory, Biotechnology Division, Department of Botany, University of Calicut, Malappuram, India.

DOI: 10.4236/abb.2012.38142                

Abstract

In this study we describe a novel dark-green strain of Trichoderma viride exhibiting complete ensemble of cellulase, hemicellulase and ligninase activities on specific plate assays. To assess the cellulase production in detail, basal salt medium (BSM) was fortified 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 137 U/mL cellulase activity, which was enhanced to 173 U/mL at 1.25% CMC concentration. Flours of potato and banana peel supported comparable yields of cellulase to that of CMC, while sodium nitrate was the preferred nitrogen source. The water soluble bluish-green pigment (a probable siderophore) extracted from the spores showed an absorption maximum at 292 nm. To sum up, the complete lignocellulolytic potential of this fungus offers great industrial significance, coupled with the production of a new pigment.

Keywords

Trichoderma viride; Morphology; Liquid Fermentation; Bluish-Green Pigment

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	Martinez, D., Larrondo, L.F., Putnam, N., Gelpke, M.D.S., Huang, K., Chapman, J., Helfenbein, K.G., Ramaiya, P., Detter, J.C., Larimer, F., Coutinho, P.M., Henrissat, B., Berka, R., Cullen, D. and Rokhsar, D. (2004) Genome sequence of the lignocellulose degrading fungus Phanerochaete chrysosporium strain RP78. Nature Biotechnology, 22, 695-700. doi:10.1038/nbt967
[2]	Rubin, E. (2008) Genomics of cellulosic biofuels. Nature, 454, 841-845. doi:10.1038/nature07190
[3]	Howard, R.L. and Abotsi, E. (2003) Issues of bioconversion and enzyme production. African Journal of Biotechnology, 2, 602-619.
[4]	Chandra, M.A., Karala, P.K., Sharma and Sangwan, R.S. (2009) Cellulase production by six Trichoderma spp., fermented on medicinal plant processings. Journal of Industrial Microbiology and Biotechnology, 36, 605-609. doi:10.1007/s10295-009-0544-9
[5]	Glick, B.R. and Pasternak, J.J. (1989) Isolation, characterization and manipulation of cellulose gene. Biotechnology Advances, 7, 361-386. doi:10.1016/0734-9750(89)90180-8
[6]	Kubicek, C.P., Komon-Zelazowska, M. and Druzhinina, I.S. (2008) Fungal genus Hypocrea Trichoderma: From barcodes to biodiversity. Journal of Zhejiang University Science, 9, 753-763. doi:10.1631/jzus.B0860015
[7]	Sánchez, C. (2009) Lignocellulosic residues: Biodegradation and bioconversion by fungi. Biotechnology Advances, 27, 185-194. doi:10.1016/j.biotechadv.2008.11.001
[8]	Gautam, S.P. (2010) Optimization of the medium for the production of cellulase by the Trichoderma viride using submerged fermentation. International Journal of Environmental Science, 1, 656-665.
[9]	Miller, G.L. (1959) Use of dinitrosalicyclic acid reagent for determination of reducing sugar. Analytical Chemistry, 31, 421-428. doi:10.1021/ac60147a030
[10]	Bissett, J. (1991) A revision of the genus Trichoderma. II. Infrageneric classification. Canadian Journal of Botany, 69, 2357-2372. doi:10.1139/b91-297
[11]	Lieckfeldt, E., Samuels, G.J., Nirenberg, H.I. and Petrini, O. (1999) A morphological and molecular perspective of Trichoderma viride: Is it one or two species? Applied and Environmental Microbiology, 65, 2418-2428.
[12]	Leonowicz, A., Matuszewska, A., Luterek, J., Ziegenhagen, D., Wojta?-Wasilewska, M., Cho, N.S., Hofrichter, M. and Rogalski, J. (1999) Biodegradation of lignin by white rot fungi. Fungal Genetics and Biology, 27, 175-185. doi:10.1006/fgbi.1999.1150
[13]	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, 559-562. doi:10.1111/j.1365-2672.1986.tb01729.x
[14]	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
[15]	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
[16]	Kang, S.W., Park, Y.S., Lee, J.S., Hong, S.I. and Kim, S.W. (2004) Production of cellulases and hemicellulases by Aspergillus niger KK2 from lignocellulosic biomass. Bioresource Technology, 91, 153-156. doi:10.1016/S0960-8524(03)00172-X
[17]	Molla, A.H., Fakhru’l-Razi, A. and Alam, M.Z. (2004) Evaluation of solid state bioconversion of domestic wastewater sludge as a promising environmental friendly disposal technique. Water Research, 38, 4143-4152. doi:10.1016/j.watres.2004.08.002
[18]	Belghith, H., Ellouz-Chaabouni, S. and Gargouri, A. (2001) Biostoning of denims by Penicillium occitanis (Pol6) cellulases. Journal of Biotechnology, 89, 257-262. doi:10.1016/S0168-1656(01)00309-1
[19]	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.
[20]	Sun, T., Liu, B.H., Li, Z.H. and Liu, D.M. (1999) Effect of air pressure amplitude on cellulase production by Trichoderma viride SL1 in periodic pressure solid state fermentation. Process Biochemistry, 34, 25-29. doi:10.1016/S0032-9592(98)00060-0
[21]	Betina, V. (1995) Photoinduced conidiation in Trichoderma viride. Folia Microbiologica, 40, 219-224. doi:10.1007/BF02814196
[22]	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.
[23]	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.