• S. T. Tyohemba
  • S. Aliyu
  • N. N. Ndukwe
  • G. G. Memi
  • U. O. Edem
Keywords: β-glucosidase, Purification, Cow dung, Enzyme activity Trichoderma viride


β-glucosidases have characteristics of biotechnological interest and have thus become important industrial enzymes.In this study, β-glucosidase produced by Trichoderma viride from cow dung was subjected to a three step purification process involving ammonium sulphate precipitation, gel filtration by Sephadex G-100 and ion exchange chromatography by DEAE-Sephadex A-25. The elution profile on Sephadex G-100 resulted in a single broad peak (fractions 9-21) which had a yield of 3.7% and a purification fold of 4.29 with a specific activity of 25.70 µmol/min/mg proteins while the elution profile on DEAE-Sephadex A-25 resulted in a single broad peak (fraction 8-14) which had a yield of 2.76% and a purification fold of 22.14 with a specific activity of 132.41µmol/min/mg of protein. The purified enzyme was obtained as a single band and had a molecular mass of 51.8 kDa on SDS-PAGE. This results provide support for further studies of this enzyme towards revealing its potential biotechnological applications.


Adegunloye, D.V., Adetuyi, F.C., Akinyosoye, F.A., and Doyeni, M.O., (2007). Microbial analysis of compost using cowdung as booster. Pakistan Journal of Nutrition, 6: 506–510.

Arja, M., John, L., Vesa, J., and Raija, L. (2004). Three cellulases from Melanocarpus albomyces for textile treatment at neutral pH. Enzyme Microbiology and Technology, 34: 332-341.

Auta, R., Wusu, A. D., Radeeka, I., and Hooley, P. (2016). Expression and characterization of recombinant β-glucosidases from Aspergillue nidulans. AN2227. Science World Journal, 11(2):1597-6343.

Bai, H., Wang, H., Sun, J., Irfan, M., Han, M., Huang, Y., Han, X. and Yang, Q. (2013). Production, purification and characterization of novel β-glucosidase from newly isolated Penicillium Simplicissimum H-11 in submerged fermentation. EXCLI Journal, 12: 528 – 540.

Binod, P., Palkhiwala, P., Gaikaiwari, R., Nampoothiri, K.M., Duggal, A., Dey, K.,

and Pandey, A. (2013). Industrial enzymes – present status and future perspectives. India Journal of Science Industrial Resources, 72: 271–286.

Bradford, M.M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical

Biochemistry, 72: 248 – 254.

Brumbauer, A., Johansson, G., Reczey, K. (2000). Study heterogeneity of β-glucosidase from Aspergillus species by using counter current distribution. Journal of Chromatography B: Biomedical Science Applied, 74:247 -254.

Chauve, M., Mathis, H., Huc, D., Casanave, D., Monot, F., and Ferreira, N.L. (2010). Comparative kinetic analysis of two fungal β–glucosidases. Biotechnology Biofuels, 3:3.

Coughlan, M.P. (1985). The properties of fungal and bacterial cellulases with comment on their production and application. Biotechnology and Genetic Engineering Reviews, 3(1): 39-110.

Del Pozo, M.V., Fernandez-Arrojo, L., Gil-Martinez, J., Montesinos, A., Chernicova, T.N., Nechitaylo, T.Y., Waliszek, A., Tortajada, M., Rojas, A., Huws, S.A., Golyshina, O.V., Newbold, C.J., Polaina, J., Ferre, M. and Golyshin, P.N. (2012). Microbial β-glucosidases from cow rumen metagenome enhance the saccharification of lignocellulose in combination with commercial cellulase cocktail. Biotechnology for Biofuels, 5(3): 1 – 13.

Dhake, A. and Patil, M. (2005) Production of ß-glucosidase by Penicillium purpurogenum. Brazilian Journal of Microbiology, 36(2): 170-176.

Han, Y.J. and Chen, H.Z. (2008). Characterization of β−glucosidase from corn stover and its application in simultaneous saccharification and fermentation. Bioresources Technology, 99: 6081-6087.

Hiol, A., Jonzo, M. D., Rugani, N., Druet, D., Sarda L., and Corneau L.C. (2000). Purification and characterization of an Extracellular Lipase from a thermophilic Rhizopus oryzae strain isolated from Palm fruit. Enzyme and Microbial Technology, 26(5-6): 421-430.

Iqbal , H. M. N., Ahmed, I., Zia, M. A., and Irfan, M. (2011a). Purification and characterization of the kinetic parameters of cellulase produced from wheat straw by Trichoderma viride under SSF and its detergent compatibility. Advances in Bioscience and Biotechnology, 2:149-156.

Iqbal, H.M.N., Asgher, M. and Bhatti, H.N. (2011b). Optimization of Physical and Nutritional Factors for Synthesis of lignin degrading enzymes by a Novel Strain of Trametes versicolor. BioResources, 6: 1273-87.

Irfan, M., Nadeem, M., and Syed Q. (2014). One-factor-at-a-time (OFAT) optimization of xylanase production from Trichoderma viride-IR05 in solid-state fermentation. Journal of Radiation Research and Applied Sciences, 7: 317–326.

Irshad M., Anwar Z., Ramzan, M., Mahmood, Z., and Nawaz, H. (2013). Characterization of purified β-glucosidase produced from Trichoderma viride through bio-processing of orange peel waste . Advances in Bioscience and Biotechnology, 4: 941-944

Jatinder, K., Bhupinder, S., Badhan, A., Ghatora, S., and Harvinder, S. (2007). Purification and characterization of β-glucosidase from Melanocarpus sp. MTCC 3922. Journal of Biotechnology, 10: 260-270.

Joo, A.R., Jeya, M., Lee, K.M., Lee, K.M, Moon, H.J., Kim, Y.S., and Lee, J.K. (2010). Production and characterization of β-1,4-glucosidase from a strain of Penicillium pinophilum. Journal Process Biochemistry, 45: 851-858.

Kaur, J., Chadha, B.S., Kumar, B.A., Kaur, G., and Saini, H.S.(2007): Purification and characterization of ß-glucosidase from Melanocarpus sp. MTCC 3922. Electronic Journal of Biotechnology, 10(2):260-270.

Laemmli, U. K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, (5259): 680-685

Misra, R. V., Roy, R. H., and Hiraoka (2003). On farm composting method. FAO, Rome.

Narasimha, G., Sridei, A., Golla, R., and Bontha, R. R (2016). Purification and characterization of β-glucosidase from A. niger. International Journal of Food Properties, 19:(3) 652-661.

Obilie, E.M., Tano-Debrah, K., and Amoa-Awua, W.K. (2004). Souring and breakdown of cyanogenic glucosides during the processing of cassava into akyeke. International Journal of Food Microbiology, 93(1): 115-121.

Saleem, M., Samiullah, T. R., Bakhsh, A., Rao, A. Q., and Naz, M. (2009). Isolation, Purification and characterization of extracellular β-glucosidases from Bacillus sp.; Advances in Environmental Biology, 3(3): 269-277.

Saloheimo, M., Nakari-Setala, T., Tenkanen, M.,and Penttila, M. (1997). cDNA cloning of a Trichoderma reesei cellulase and demonstration of endoglucanase activity by expression in yeast. European Journal of Biochemistry, 249: 584-591.

Sorensen, A., Lubeck, M., Lubeck, P.S., and Ahring, B.K. (2013). Fungal betaglucosidases: a bottleneck in industrial use of Lignocellulosic materials. Biomolecules, 3(3):612 –631.

Swangkeaw, J., Vichitphan, S., Butzke, C.E., and Vichitphan , K.(2009). The characterisation of a novel Pichia anomala β-glucosidase with potentially aroma-enhancing capabilities in wine Annals of Microbiology, 59 (2): 335-343.

Tomaz, T. and Roche, A. (2002). Hydrophobic interaction, Chromatography of Trichoderma reesei cellulase on Polypropylene glycol-Sepharose. Separation Science Technology, 37: 1-11.

Tsao, G. T., Xia, L., Cad, N., and Gong,C.S. (2000). Solid state fermentation with Aspergilus niger for cellobiase production. Applied Biochemistry and Biotechnology, 84-86:743-747.

Viniegra-Gonzalez, G., and Favela-Torres, E (2006). Why solid-state fermentation seems to be resistant to catabolite repression. Food Technology and Biotechnology, 44 (3): 397 – 406.

Zhang, C., Li, D., Yu, H., Zhang, B., Jin, F. (2007). Purification and characterization of Peceid β-glucosidase from Aspergillius orgzae. Press Biochemistry, 42:83-88.

How to Cite
TyohembaS. T., AliyuS., NdukweN. N., MemiG. G., & EdemU. O. (2020). PURIFICATION OF β-GLUCOSIDASE PRODUCED FROM Trichoderma viride USING COW DUNG AS CARBON SOURCE. FUDMA JOURNAL OF SCIENCES, 4(2), 190 196. https://doi.org/10.33003/fjs-2020-0402-210

Most read articles by the same author(s)