Investigating of the Influence of Cellulase Enzymes from Mutated Isolates of Trichoderma harzianum and T. viride on Biodegradation of Cellulose Iα, Iβ and III

Document Type : research

Authors

1 Assistant Professor , Plant Protection and Food Preservation Department, Nuclear Agriculture Research School, Nuclear Science and Technology Research Institute (NSTRI), Atomic Energy Organization of Iran (AEOI), Alborz, Iran

2 MSc Graduated, Plant Protection and Food Preservation Department, Nuclear Agriculture Research School, Nuclear Science and Technology Research Institute (NSTRI), Atomic Energy Organization of Iran (AEOI), Alborz, Iran

Abstract

In this research,two strain of Trichoderma (T. harzianum and T. viride) and their mutants were used for cellulase enzyme production. The Avicel, CMC(III), Bacterial Cellulose (Iα) and Whatman NO.1 filter paper, were used for cellulase activity assay. Their chemical structural properties were investigated by FT-IR, XRD and SEM compared to Avicel. The molecular weight of cellulase enzymes were studied using SDS-PAGE. The SEM image of substrates, showed more delicacy of BC fibers relative to Avicel and CMC. The diameter ratio of BC to Avicel is approximately 1/30 or less. The FTIR spectroscopy and XRD assessments designated that the produced CMC is carrying a carboxylic group and its crystallinity is 81.84. The crystallinity of the Avicel and BC were demonstrated that the crystallinity index of Avicel (89.15%) was more than that of bacterial cellulose (66.44%). Both species and their mutants produced varying amounts of extracellular proteins in the fermentation medium. In T. harzianum and its mutants, the highest cellulase enzyme activity were showed in Th M7 and Th M6, respectively and in T. viride and its mutants, the highest enzymatic activity of mutant strains was observed in Tv M14, Tv M15, respectively. The results showed that bacterial cellulose is a good substrate for total cellulase activity assay, because it has the highest enzyme activity compared to filter paper and avicel; so it signs a high potential of accessibility for cellulose Iα biodegradability compared to cellulose Iβ.

Keywords

References

ASTM. 2005. Analytical Method for Determining Degree of Substitution in the Product, Document CK-G06 Edition, 05 D-1439-03.
Bradford, M. M., 1976. A rapid and sensitive for the quantitation of microgram of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 72: 248-258.
Cannon, R. E. and Anderson, S. M. 1991. Biogenesis of bacterial cellulose. Critical Reviews in Microbiology, 17(6): 435-447.
de Palma-Fernandez, E. R., Gomes, E. and Da Silva, R. 2002. Purification and characterization of two β-glucosidases from the thermophilic fungusThermoascus aurantiacus. Folia microbiologica, 47(6): 685-690.
Foreman, P. K., Brown, D., Dankmeyer, L., Dean, R., Diener, S., DunnColeman, N. S., Goedegebuur, F., Houfek, T. D., England, G. J., Kelly, A. S., Meerman, H. J., Mitchell, T., Mitchinson, C., Olivares, H. A., Teunissen, P. J. M., Yao, J. and Ward, M., 2003. Transcriptional regulation of biomass-degrading enzymes in the filamentous fungus Trichoderma reesei. Journal of Biology and Chemistry, 278: 31988-31997.
Gama, F. M. and Mota, M. 1998. Cellulases for oligosaccharide synthesis: a preliminary study. Carbohydrate Polymers, 37: 279-281.
Grishutin, S. G., Gusakov, A. V., Markov, A. V., Ustinov, B. B., Semenova, M. V. and Sinitsyn, A. P. 2004. Specific xyloglucanases as a new class of polysaccharide-degrading enzymes. Biochimica et Biophysica Acta (BBA)-General Subjects, 1674(3): 268-281.
Haigler, C. H. and Weimer, P. J. 1991. Biosynthesis and Biodegradable of Cellulose. New York: M. Dekker, 694pp.
Karlsson, J., Saloheimo, M., Siika-aho, M., Tenkanen, M., Penttilä, M. and Tjerneld, F. 2001. Homologous expression and characterization of Cel61A (EG IV) of Trichoderma reesei. European Journal of Biochemistry, 268(24): 6498-6507.
Kirk, R. E. and Othmer, D. F. 1967. Cellulose (2nd ed.). Encylopedia of chemical technology,Vol. 4. New York: Wiley, pp: 593-683.
Lynd, L. R., Weimer, P. J., Van Zyl, W. H. and Pretorius, I. S. 2002. Microbial cellulose utilization: fundamentals and biotechnology. Microbiology and Molecular Biology Reviews, 66(3): 506-577.
Maki, M., Leung, K. T. and Qin, W. 2009. The prospects of cellulase-producing bacteria for the bioconversion of lignocellulosic biomass. International Journal of Biological Science, 5: 500-516.
Moosavi-Nasab, M., Yousefi, A. R., Askari, H. and Bakhtiyari, M. 2010. Fermentative production and characterization of carboxymethyl bacterial cellulose using date syrup. World Academic Science Engineering Technology, 68: 1467-1471.
Moradi, R., Shahbazi, S., Ahari mostafavi, H., Ebrahimi, M. A., Askari, H. and Mirmajlesi, M. 2015. Investigation of gamma radiation effects on morphological and antagonistic characteristics of Trichoderma harzianum. Journal of Nuclear Science and Technology, 71: 96-104.
Nidetzky, B. and Claeyssens, M. 1994. Specific quantification of Trichoderma reesei cellulases in reconstituted mixtures and its application to cellulase-cellulose binding studies. Biotechnology Bioengineering, 44: 961-966.
Ross, P., Mayer, R. and Benziman, M. 1991. Cellulose biosynthesis and function in bacteria. Microbiological Reviews, 55(1): 35-58.
Sajith, S., Priji, P., Sreedevi, S. and Benjamin, S. 2016. An overview on fungal cellulases with an industrial perspective. Journal of Nutral Food Science, 6: 461. doi:10.4172/2155-9600.1000461.
Schülein, M. 1997. Enzymatic properties of cellulases from Humicola insolens. Journal of Biotechnology, 57(1-3): 71-81.
Shahbazi, S., Askari, H. and Mojerlou, S., 2016. The impact of different physicochemical parameters of fermentation on extracellular cellulolytic enzyme production by Trichoderma harzianum. Journal of Crop Protection, 5(3): 397-412.
Shahbazi, S., Askari, H., Naseripour, T., Moosavi-Nasab, M. and Bakhtiyari, M. 2014. The synergistic interactions of Trichoderma spp. cellulase enzyme activities in biomass conversion; Part1: comparison of cellulose Iα, Iβ and III. International Journal of Agriculture and Crop Sciences (IJACS), 7(8): 442-453.
Sugiyama, J., Vuong, R. and Chanzy, H. 1991. Electron diffraction study on the two crystalline phases occurring in native cellulose from an algal cell wall. Macromolecules, 24(14): 4168-4175.
Teeri, T. and Koivula, A. 1995. Cellulose degradation by native and engineered fungal cellulases. Carbohydrates in Europe, 12: 28-33.
Wen, Z., Liao, W. and Chen Sh. 2005. Production of cellulase by Trichoderma reesei from dairy manure. Bioresource Technology, 96: 491-499.
Yan, Z., Chen, S., Wang, H., Wang, B., Wang, C. and Jiang, J. 2008. Cellulose synthesized by Acetobacter xylinum in the presence of multi-walled carbon nanotubes. Carbohydrate Research, 343: 73-80.
Zhang, Y. H. P. and Lynd, L. R. 2006. A functionally based model for hydrolysis of cellulose by fungal cellulase. you have free access to this content. Biotechnology and Bioengineering, 94(5): 888-898.
Volume 9, Issue 1
June 2018
Pages 35-44

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