Cellulase Induction in Three Aspergillus Species Isolated From Artemisia annua L. Plantation Soil Using Different Cellulose Substrates

A. I. Ogbonna, P. O. Nwadiaro, A. Chuku, C. I. C. Ogbonna, F. C. Onwuliri

Abstract


This study was aimed at isolation and screening of fungal species associated with Artemisia annua Plantation soils from one of the under studied areas in Plateau state, Nigeria for cellulase activity. A total of thirteen fungal species were isolated from various locations within the A. annua Plantation and were screened for cellulase production. Agar plate assay was carried out using basal medium supplemented with 1% Carboxymethylcellulose (CMC) powder and staining with 0.1% Congo red solution after the incubation period. Among these species, Aspergillus fumigatus, A. niger and A. terreus were more predominant and were recorded as cellulase producing species. They have shown to possess cellulose degrading ability and exhibited maximum zones of hydrolysis on Carboxymethylcellulose medium and were selected for enzyme assay using submerged fermentation (SmF). Enzyme production was analyzed by Dinitrosalicylic acid (DNSA) methods and the enzymes assayed for were CMCase (β-1,4-endoglucanase), β-glucosidase and FPase (total cellulose) using Carboxymethylcellulose, cellulose acetate and Filter paper as substrates respectively. The highest cellulase activity was observed on the 3rd day in A. niger with enzyme production of 0.045IU/ml and 0.040IU/ml on CMC and filter paper media respectively. A. fumigatus had high enzyme activity of 0.037IU/ml and 0.025IU/ml on filter paper and cellulose acetate media respectively. Highest enzyme production of 0.034IU/ml was recorded for A. terreus on the 3rd day on cellulose acetate medium. These fungal species could be employed specially to perform in situ environmental applications involving cellulose biodegradation of wastes.


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References


Akiba S, Kimura Y, Yamamoto K, Kumagai H (1995). Purification and characterizationof a protease-resistant cellulase from Aspergillus niger. J. Ferment. Bioeng. 79(2):125-130.

Ali S, Sayed A, Sarker RI, Alam R (1991). Factors affecting cellulose production by Aspergillus terreus using water hyacinth. World J. Microbiol and Biotechnol 7:62-66.

Anita, B. B., Thatheyus, A.J. and Vidhya, R. (2013). Biodegradation of Carboxymethyl Cellulose Employing Cheatomium globosum MTCC 2193. Science International, 1: 92-97.

Barnett H.L. and Hunter B.B. (1972) Illustrated genera of Imperfect Fungi, Burgress Publishing Company, Minneapolis, Minnesota, USA. 241 pp.

Chellapandi, P., Himanshu, M. J. (2008) Production of endoglucanase by the native strains of Streptomyces isolates in submerged fermentation. Brazilian J Microbiol. 39: 122-127. http://dx.doi.org/10.1590/S1517-83822008000100026

Chinedu, S.N., O.C. Nwinyi and V.I. Okochi, 2008. Growth and cellulose activity of wild-type Aspergillus niger ANL301 in different carbon sources. Can. J. Pure Appl. Sci., 2: 357-362.

Coral G, Arikan B, Unaldi MN, Guvenmes H (2002). Some properties of crude carboxymethyl cellulase of Aspergillus niger Z10 wild-ype Strain. Turk J. Biol. 26:209-213.

Gielkens MMC, Dekkers E, Visser J, Graaff L H (1999). Two cellubiohydrolase-encoding genes from Aspergillus niger require D Xylose and the xylanolytic transcriptional activator XlnR for their expression. Appl. Environ. Microbiol. 65(10):4340-4345.

Hamilton, J.A., and Was, D. (1991) Some components of cellulases from two different strains of Aspergillus fumigatus. Process Biochem. 26: 287-290.

Hong J, Tamaki H, Akiba S, Yamamoto K, Kumaga H (2001). Cloning of a gene encoding a highly stable endo1,4-glucanase from Aspergillus niger and Its expression in Yeast. J. Biosci. Bioeng. 92(5):434-441.

Khalid, M., Yang, W.J., Kishwar, N., Rajput, Z.I. & Arijo, A.G. 2006. Study of cellulolytic soil fungi and two nova species and new medium. J. Zhejiang Univ. Sci. B, 7(6): 459-466.

Kluepfel D, Shareck F, Mondou F, Morosoli R (1986). Characterization of cellulose and xylanase activities of Streptomyces lividans. Applied Microbiol. Biotechnol. 24:230-234.

Miller, G. L. (1959). Use of Dinitrosalicyclic reagent for the determination of reducing sugars. Anal. Chem., 31: 426-428.

Mukunda S, Onkarappa R, Prashith Kekuda T. R. (2012). Isolation and Screening of Industrially Important Fungi from the Soils of Western Ghats of Agumbe and Koppa, Karnataka, India. Science, Technology and Arts Research Journal 1(4):27-32.

Onion, A H S; Allsop, D; Eggins, H O W (1981). Smiths Introduction to Industrial Mycology (7th Ed.) Edward-Arnold Publishers Ltd. London p.398.

Oyeyiola, G .P., Hussein, H. S. N. (1992). Fungi populations in the rhizosphere and rhizoplane of wheat grown in Kura Area of Kan, Northern Nigeria. Bioscience Research Communications 4(2): pp:129-133.

Peciulyte, D., 2007. Isolation of cellulolytic fungi from waste paper gradual recycling materials. Ekologija, 53: 11-18.

Pointing, S.B. (1999). Qualitative methods for the determination of lignocellulolytic enzyme production by tropical fungi. Fungal Diversity. 2: 17-33.

Rosgaard, L., Pedersen, S., Cherry, J. R., Harris, P., Meyer, A. S. (2006). Efficiency of new fungal cellulose systems in boosting enzymatic degradation of barley straw lignocelluloses. Biotechnol Prog. 22 (2): 493-498.

Sathyaprabha, G., Panneerselvam, A and Muthukkumarasamy, S. (2011). Production of Cellulase and Amylase from wild and mutated fungal isolates. E-Journal of Life Sciences 1(1): 39-45.

Sharma TR, Sreekantiah KR (1986). Production of cellulases and D-Xylanase by some selected fungal isolates. Enzyme Microbiol Technol. 8:178-182.

Sivakumaran, S. (2014). Isolation of Cellulolytic Fungi and their Degradation on Cellulosic Agricultural Wastes 2 (8) 458-463.

Steiner J, Socha C, Eyzaguirre J. (1994) Culture conditions for enhanced cellulase production by a native strain of Penicillium purpurogen. World J Microbiol Biotechnol. 10: 280-284.

Szakacs G, Boagar B, Molnar T, Szilagi E, Fekete J, Tengerdy RP (2001). Optimization of α- amylase and lovasatin production in solid substrate fermentation; In: Intl Conf. on New Horizons in Biotechnol ,Trivandram, India. 18-21.

Uttam, K., Ashwani, T., Pinky, K., Shiju, V., Suresh, C. (2014). Isolation and Screening of Cellulase Producing Fungi from Forest Waste. International Journal of Pharmaceutical & Biological Archives 5(1): 56 - 59.

Von Arx, J.A. (1974). The Genera of Fungi sporulating in pure culture Publ. J. Grammar. In der A.R. Gantner Verlog Kommanditesells chaft, Fl- 9490 Vaduz, Germany 315 pp.

Warcup, J. H. (1950).The soil plate method for isolation of fungi from soil. Nature 166:117-118.

Wilson, D.B. and Irwin, D.C. (1999). Genetics and properties of cellulases. Advances in Biochemical Engineering/ Bio-technology: Recent Progress in Bioconversion 65: 1-21.

Wood TM, Garcia-Campayo V. Enzymology of cellulose degradation. Biodegradation 1990; 1: 147- 161.




DOI: https://doi.org/10.5296/jab.v3i1.6738

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