Purification and Partial Characterization of Cellulase Produced by Aspergillus niger Cultured on Vitellaria paradoxa shells

A. O. Sulyman; Y. A. Iyanda; A. O. Opasola; O. Adedayo; R. A. Aladodo; A. Balogun; O. A. Ajibola; O. Z. Olutokun; B. A. Hammed

doi:10.53704/fujnas.v6i2.167

Authors

A. O. Sulyman Department of Biochemistry, College of Pure and Applied Sciences, Kwara State University, Malete
Y. A. Iyanda Department of Environmental Management and Toxicology, Kwara State University, Malete, Ilorin, Nigeria
A. O. Opasola Department of Environmental Management and Toxicology, Kwara State University, Malete, Ilorin, Nigeria
O. Adedayo Department of Biosciences and Biotechnology (Microbiology unit), Kwara State University, Malete, Ilorin, Nigeria
R. A. Aladodo Department of Biochemistry, Kwara State University, Malete, Ilorin, Nigeria
A. Balogun Department of Biological Science (Biochemistry Unit), Al-Hikmah University, Ilorin, Nigeria
O. A. Ajibola Department of Biological Science (Biochemistry Unit), Al-Hikmah University, Ilorin, Nigeria
O. Z. Olutokun Department of Biological Science (Biochemistry Unit), Al-Hikmah University, Ilorin, Nigeria
B. A. Hammed Department of Biological Science (Biochemistry Unit), Al-Hikmah University, Ilorin, Nigeria

DOI:

https://doi.org/10.53704/fujnas.v6i2.167

Abstract

This research investigated the purification and partial characterization of cellulase produced by Aspergillus niger cultured on Vitellaria paradoxa shells. Cellulase (endoglucanase) from A. niger was produced under optimum fermentation conditions at 35 °C, pH 4.7, V. paradoxa, 4 g/L, inoculum size of 10 mm and the fermentation media incubated for 120 hours. The crude endoglucanase obtained were partially purified by subjecting to ammonium sulphate precipitation, dialysis and gel filtration chromatography for further purification. The effect of temperature and pH on the activity of purified endoglucanase was determined. Cellulase was purified to 734.33 folds by Sephadex G-100 column chromatography with a specific activity and yield of 4.406 U/mg and 63.03% respectively. Fractions 4 and 7 contained the highest endoglucanase activity out of 18 fractions collected and the two fractions were pooled for further analysis. The activity of purified endoglucanase was optimum at a temperature of 40 °C and pH 5. Therefore, the purified endoglucanase produced may be explored in detergent industry.

Keywords: Aspergillus niger, endoglucanase, Vitellaria paradoxa, purification, partial characterization

References

Abagale, F.K., Abdulai, N. and Ojediran, J.O. (2012). Effects of Shea waste slurry on soil physical properties in Peri-Urban Tamale, Northern Ghana. Asian Journal of Science and Technology, 4(12): 036-041.

Acharya, P. B., Acharya, D. K. and Modi, H. A. (2008). Optimization for cellulase production by Aspergillus niger using sawdust as substrate. African Journal of Biotechnology, 7(22): 4147–4152.

Adrio, J. L. and Demain, A. L. (2014). Microbial enzymes: tools for biotechnological processes. Biomolecules, 4(1): 117-139.

Ajayi, A.O. (2004). Emerging roles for extension in promoting sustainable rural environment: lessons from food processing cottage industries and their wastes in rural Oyo State, Nigeria. Journal of Human Ecology, 16(4): 283-289.

Alander, J. (2004). Shea butter – a multifunctional ingredient for food and cosmetics. Lipid Technology 16(9): 202-205

Amadioha, A. C. (1993). Production of cellulolytic enzymes by Rhizopus oryzae in culture and Rhizopus-infected tissues of potato tubers. Mycologia, 85(4): 574-578.

Amin, M., Bhatti, H. N., Zuber, M., Bhatti, I. A. and Asgher, M. (2014). Potential use of agricultural wastes for the production of lipase by Aspergillus melleus under solid state fermentation. Journal of Animal and Plant Science, 24(5): 1430-1437.

Arvanitoyannis, I.S. and Varzakas, H.S. (2008). Vegetable waste treatment: Comparison comparison and critical presentation of methodologies. Critical Reviews in Food Science and Nutrition, 48: 205-247.

Belghith, H., Elbouz-Chaobouni, S. and Gargouri A. (2001). Biostoning of denims by Penicillum acetanis, vol.6; cellulose. Journal of Biotechnology, 6: 257-262

Bhadauria, A., Sodhi, H. S., Kapoor, S., and Phutela, R. P. (1997). Isolation and characterization of Volvariella diplasia enzyme mutants. Indian Journal of Experimental Biology, 35(4): 516-519.

Bhavna, V.M. and Magar, J.G. (2010). Use of agricultural wastes for cellulase production by Aspergilus niger with submerged and sold state fermentation; Bionano Frontier, 3 (2): 189-192

Cavazzoni, V., and Manzoni, M. (1994). Extracellular cellulolytic complex from Morchella conica: production and purification. LWT-Food Science and Technology, 27(1): 73-77.

Chen, P. J., Wei, T. C., Chang, Y. T., and Lin, L. P. (2004). Purification and characterization of carboxymethyl cellulase from Sinorhizobium fredii. Botanical Bulletin of Academia Sinica, 45. 111-118

De Azeredo, L.A.I., Gomes, P.M., Sant’Anna, G.L., Castilho, L.R. and Freire, D.M. (2007). Production and regulation of lipase activity from Penicillium restrictum in submerged and solid-state fermentations. Current Microbiology. 54: 361–365

De-Moraes, L.M.P., Filho, S.A. and Ulhaa, C.J. (1999). Purification and some properties of an alpha amylase and glucoamylase fusion protein from Saccharomyces cerevisiae. World Journal of Microbiology and Biotechnology, 15: 561-564.

Food and Agricultural Organization, (2005). Production Year Book, Trade, 46(115): 18-24

Garba, I.D., Nwawe, C.N. and Oisakede, I.L. (2011). The potentials of Shea nut tree to the Nigerian Economy. International Journal of Agricultural Economics and Rural Development, 4: 62-72.

Ghose, T. K. (1987). Measurement of cellulase activities. Pure and Applied Chemistry, 59(2): 257-268.

Goma, M., Zein, G. N., Mahmoud, R. M., Gibriel, A. and Abouzied, M. (1982). Characteristics of cellulolytic enzymes of Aspergillus niger and Aspergillus terreus. Minufiya Journal of Agricultural Research, 5: 299-317.

Gupta, J. K., and Gupta, Y. P. (1979). Properties of cellulase from Trichoderma viride. Folia microbiologica, 24 (3): 269-272.

Gusakov, A. V., Berlin, A. G., Popova, N. N., Okunev, O. N., Sinitsyna, O. A., and Sinitsyn, A. P. (2000). A comparative study of different cellulase preparations in the enzymatic treatment of cotton fabrics. Applied Biochemistry and Biotechnology, 88(1-3): 119-126.

Immanuel, G., Akila-Bhagwat C.M., Iyappa-Raj, P., Esakkiraj, P. and Palavesam A. (2009). Production and Partial purification of Cellulase by Aspergillus niger and A. fumigates fermented in coir waste and sawdust, Internetational Journal of Microbiology, 3(1): 1-20.

IPGRI, INIA, (2006). Descriptors for Shea Tree (Vitellaria paradoxa). International Plant Resource Institute, Rome, Italy.

Jibreel, M.B., Mumuni, E., Al-Hassan, S. and Baba, N.M. (2013) Shea butter and its processing impacts on the environment in the Tamale Metropolis of Ghana. International Journal of Development and Sustainability, 2(3): 2008-2019.

Kottwiz, B. and Schambil, F. (2005) Cellulase and Cellulase containing detergents, Us pat 20050020472

Lee, K. C., Tong, W. Y., Ibrahim, D., Arai, T., Murata, Y., Mori, Y. and Kosugi, A. (2017). Evaluation of enzymatic deinking of non-impact ink laser-printed paper using crude enzyme from Penicillium rolfsii c3-2 (1) IBRL. Applied Biochemistry and Biotechnology, 181(1): 451-463.

Lovett, P. N. (2004). The Shea butter value chain: production, transformation and marketing in West Africa. West Africa Trade Hub (WATH) Technical Report No. 2. US-AID West Africa Programme.

Lowry, O., Rosebrough, N., Farr, A. and Randall, R. (1951). Protein measurement with Folin-phenol reagent. Journal of Biological Chemistry, 193: 265275.

Macris, B. J., Kekos, D., Evangelidou, X., Galiotou-Panayotou, M., and Sodis, P. (1987). Solid state fermentation of straw with Neurospora crassa for CMCase and ?-glucosidase production. Biotechnology Letters, 9 (9): 661-664.

Mishra, A. (1988). Studies a cellulolytic fungi and cellulolytic enzymes with special reference to Aspergilli (Doctoral dissertation, Ph. D. Thesis, Department of Botany, Gorakhpur Univ. India.

Olama, Z. A., Hamza, M. A., El-Sayed, M. M., and Abdel-Fattah, M. (1993). Purification, properties and factors affecting the activity of Trichoderma viride cellulase. Food Chemistry, 47(3): 221-226.

Po-Jui, C., TaoChun, W., YaoTsung, C. and Liangling, L. (2004). Purification and characterization of carboxylmethyl cellulase from Sinorhigobium fredii. Botanical Bulletin - Academia Sinica. 45: 111-118.

Rousseau, K., Gautier, D. and Wardell, D.A. (2015). Coping with the upheavals of globalization in the Shea value chain: the maintenance and relevance of upstream Shea nut supply chain organization in western Burkina Faso. World Development, 66: 413–427

Sandhya, S. (1992). Effect of temperature on deactivation of cellulase. Asian Environment, 78-81.

Sarkodie, P.A., Agyapong, D.I., Mumuni S. and Amponsah, F.Y. (2016). Assessing the impact of indigenous shea butter processing activities in northern Ghana. International Research Journal of Environment Sciences, Vol. 5(3): 18-26.

Sharma, J., Singh, A., Kumar, R., and Mittal, A. (2006). Partial purification of an alkaline protease from a new strain of Aspergillus oryzae AWT 20 and its enhanced stabilization in entrapped Ca-Alginate beads. Internet Journal of Microbiology, 2(2): 1-14.

Shen, F., Smith Jr, R. L., Li, L., Yan, L., and Qi, X. (2017). Eco-friendly method for efficient conversion of cellulose into levulinic acid in pure water with cellulase-mimetic solid acid catalyst. ACS Sustainable Chemistry and Engineering, 5(3): 2421-2427.

Shin, C.S., Lee, J.P., lee, I.S. and Park, S.C., (2000). Enzyme production of Trichoderma ressei, Rut C-30 on various lignocellulosic substrates, Applied Biochemistry and Biotechnology, 84-86 (1-9): 237–245

Shitu, S., and Mohd-Nor, R. (2016). Shea Kernel Supply Chain and Suppliers Relationships in Rural Borgu, Niger State, Nigeria. Imperial Journal of Interdisciplinary Research, 2 (5). 427-433

Sukumaran, R. K., Singhania, R. R. and Pandey, A. (2005). Microbial cellulases: production, applications and challenges. Journal of Scientific and Industrial Research, 64: 832-844.

Sultana, S. (1997). Isolation of cellulolytic microorganisms and their activities (Doctoral dissertation, M. Phil. Thesis, Institute of Biological Science. University of Rajshahi, Bangladesh).

Sulyman, A. O., Iyanda, Y. A., Balogun, A., Ahmad, J. B., Aladodo, R. A., Aliyu, R. and Ajadi, R. O. (2018). Effect of pH and temperature on the activity of lipase produced by Aspergillus niger cultured on Citrus sinensis peel. Journal of Biological and Chemical Research, 35 (1): 181-187.

Wang, J. S. (1999). Cellulase production by a mutant strain of Trichoderma reesei from bagasse. In 23rd ISSCT Congress, New Delhi, India, (pp. 67-76).

Warra, A. A., (2011). Cosmetic potentials of African Shea nut (Vitellaria paradoxa) butter. Current Research in Chemistry, 3(2): 80-86.

Yuangklang, C., Schonewille, J. T., Alhaidary, A., Vasupen, K., Bureenok, S., Seanmahayak, B. and Beynen, A. C. (2017). Growth performance and macronutrient digestion in goats fed a rice straw-based ration supplemented with fibrolytic enzymes. Small Ruminant Research, 154: 20-22.