Optimization Studies on Cellulase Production by Aspergillus niger and Aspergillus fumigatus
Asian Journal of Biotechnology and Genetic Engineering,
Aim: In this study, two fungal species, Aspergillus niger and Aspergillus fumigatus were screened and optimized for their abilities to degrade cellulose using filter paper and Carboxymethylcellulose (CMC) as substrates.
Materials and Methods: Aspergillus niger and A. fumigatus procured from the Applied Microbiology Unit of Department of Plant Science and Biotechnology, University of Jos were screened using Whatman No. 1 filter paper and Carboxymethylcellulose as substrates in Petri plates. The fungal species abilities to produce cellulase at varying optimization parameters such as incubation periods (5 days), different incubation temperatures (25-500C), different pH(3-9) and different substrate concentration (0.25-2%) using Submerged Fermentation (SmF) were also studied.
Results: The plate assay showed that the two species produced cellulases. The highest cellulolytic activity was shown by A. niger (23±3.22 mm) as it had larger zones of clearance compared to A. fumigatus (13±3.06 mm). However, the organisms grown on filter paper agar showed better hydrolysis compared to the growth on CMC agar. For the Submerged Fermentation (SmF), enzyme activity increased for the first 98 hours of incubation on filter paper recording 2.62 IU/ml for A. niger and 2.45 IU/ml for A. fumigatus after 48 h and then there was decrease in enzyme activity. For the CMC, the highest enzyme activity was observed at 48 h recording 1.76 U/ml and 1.37 IU/ml for A. niger and A. fumigatus respectively. Maximum enzyme production was observed at incubation temperature of 30 0C for A. niger and A. fumigatus recording 1.05 IU/ml and 1.10 IU/ml on filter paper. Enzyme activity was found to be highest at pH 6 with A. niger and A. fumigatus recording 2.27 IU/ml and 2.03 IU/ml respectively on CMC broth. The 2% substrate concentration gave the highest enzyme activity of 0.58IU/ml and 0.54IU/ml for A. niger and A. fumigatus respectively. The increase was linear, the higher the concentration of the substrate, the higher the enzyme activity.
Conclusion: Aspergillus niger and A. fumigatus have demonstrated potential of synthesizing hydrolytic cellulolytic enzymes and could be employed in the degradation of lignocellulosic wastes. These enzymes could find applications in different industries.
- Cellulolytic fungi
- filter paper
- Carboxymethyl cellulose
- Congo red
How to Cite
Abu EA, Onyenekwe PC, Ameh DA, Agbaji AS, Ado SA. Cellulase (EC 220.127.116.11) production from sorghum bran by Aspergillus niger SL1: An assessment of pre-treatment methods. Proceedings of the International Conference on Biotechnology: Commercialization and Food Security, Abuja, Nigeria, 2000;153-157.
Zhang YH, Himmel ME, Mielenz JR. Outlook for cellulase improvement: Screening and Selection Strategies. Biotechnol. Adv. 2006; 24: 452-481.
Spence KL, Venditti RA, Habibi Y, Rojas OJ, Pawlak JJ. The effect of chemical composition on microfibrillar cellulose films from wood pulps: mechanical processing and physical properties. Bioresour. Technol. 2010;101:5961-5968.
Filson PB, Dawson-Andoh BE Sono-chemical preparation of cellulose nanocrystals from lignocellulose derived materials. Bioresour. Technol. 2009;100: 2259-2264.
Jahangeer Sadaf, Khan Nazia, Jahangeer Saman, Sohail Muhammad, Shahzad Saleem, Ahmad Aqeel, Khan Ahmed Shakeel. Screening and Characterization of Fungal Cellulases Isolated from the Native Environmental Source. Pak. J. Bot. 2005; 37(3): 739-748.
Miettinen-Oinonen A, Londesborough J, Joutsjoki V, Lantto R, Vehmaanperä J. Three cellulases from Melanocarpus albomyces with applications in the textile industry. Enz Microbial Technol. 2004; 34: 332 341.
Cavaco-Paulo A, Gübitz G. Catalysis and processing. In: Cavaco-Paulo A, Gübitz G. (ed.). Textile Processing with Enzymes. England, Woodhead Publishing Ltd: 2003; 86-119.
Walsh G. Industrial enzymes: proteases and carbohydrases. In: Proteins; Biochemistry and Biotechnology. John Wiley and Sons. Ltd.; 2002.
Peciulyte D. Isolation of Cellulolytic fungi from waste paper gradual recycling materials. Ekologija 2007; 53: 11-18.
Davis B. Factors influencing protoplast isolation. In: Peberdy J. F., Ferenizy L. (eds.). Fungal Protoplasts: Applications in Biochemistry and Genetics. Marcel Dekker, New York: 1985; p356.
Mandels ML, Hontz L, Nistrom J. Enzymatic hydrolysis of waste cellulose. Biotechnol Bioengineering. 1974; 16: 471–493.
Bhat MK. Cellulases and related enzymes in biotechnology. Biotechnol Adv. 2000; 18(5):355-83.
Lo C, Zhang Q, Lee P, Ju L. Cellulase production by Trichoderma reesei using sawdust hydrolysate. Applied Biochem. Biotechnol. 2005; 121-124: 561-573.
Betini JHA, Michelin M, Peixoto-Noggueira SC, Jorge JA, Terenzi HF, Polizeli MLTM.. Xylanases from Aspergillus niger, Aspergilus niveus and Aspergillus ochraceus Produced under Solid State Fermentation and their Application in Cellulose pulp bleaching. Bio proc. Biosyst. Eng, 2009; 32: 819-824.
Nagamani A, Kunwar IK, Manoharachary C. Handbook of soil fungi. IK International Pvt. Ltd., New Delhi. 2006;475.
Domsch KH, Gams W, Anderson TH. Compendium of Soil Fungi. London, New York, Torroute, Sydney, San Francisco: Academic Press. 1980; 869pp.
Jeffries TW. Manual on production and applications of cellulose laboratory Procedures . 1996;1-10.
Ja’afaru MI, Fagade OE. Optimization studies on cellulase enzyme production by an isolated strain of Aspergillus niger YL128. African Journal of Microbiology Research2010; (24): 2635-2639.
Corel G, Anikan B, Unaldi MN, Guvenmez H. Some properties of crude Carboxymethylcellulase of Aspergillus niger Z10 wild type strain TUBITAK. 2002; 26: 206-213.
Ojumu TV, Solomon OB Betiku E, Layokun SK Amigun B. Cellulase production by Aspergillus flavus Linn. isolate NSPR 101 fermented in sawdust, bagasse and corncob. Afr. J. Biotechnol. 2003; 2: 150-152.
Mandal M, Ghosh U. Value Addition to Horticultural Solid Waste by Applying It in Biosynthesis of Industrially Important Enzyme: Cellulase. In Utilization and Management of Bioresources 2018; pp. 279-289.
Iqbal HMN, Ahmed I, Zia MA, Irfan M. Purification and characterization of the kinetic parameters of cellulase produced from wheat straw by Trichoderma viride under SSF and its detergent compatibility. Adv. Biosci. Biotechnol. 2011; 2: 149-56.
Sun H, Ge X, Hao Z, Peng M. Cellulase production by Trichoderma sp. on apple pomace under solid state fermentation. Afr. J. Biotechnol 2010; 9(2): 163-166.
Nochure SV, Roberts MF Demain AL. True cellulase production by Clostridium thermocellum grown on different carbon sources. Biotech. Lett. 1993;15(6):641-646.
Sonjoy SB, Bex K, Honston H. Cellulose Activity of Trichoderma reesei (rut 30) on Municipal Solid Waste. Applied Biochemistry and Biotechnology. 1995; 51-52(1): 145-153.
Devanathan G, Shanmugan A, Balsibramanian T, Manivannan, S. (2007). Cellulase Production by Aspergillus niger Isolated from coastal Mangrove Debris. Trends in Applied Science Resources, 2: 23-27.
Acharya PB, Acharya DK, Modi HA. Optimization for cellulose production by Apsergillus niger using sawdust as substrate. African Journal of Biotechnology, 2008; 7(22): 4146-4152.
Sherif AA, El-Tanash AB, Atia, N. Cellulase production by Aspergillus fumigatus grown on mixed substrate of rice straw and wheat Bran. Research Journal of Microbiology 2010; 5(3): 199-211.
Hanif A, Tasmeen A, Rajoka MI. Induction, Production, Repression and Expression of exoglucanase synthesis in Aspergillus niger. Bioresource Technology. 2004; 94 (3): 311-319.
Deswal D, Khasa TP, Kuhad RC. Optimization of Cellulase Production by a brown not fungus fomitopsis sp. RCK 2010 under solid state fermentation. Bioresoruce Technology. 2011; 102: 6065-6072.
Irshad M, Asgher M, Sheikh MA, Nawaz H. Purification and characterization of lactase produced by Schyzophylum commune IBL-06 in solid state culture of banana stalks. Bio Res 2011; 6: 2861-2873.
Dutta T, Sahoo R, Sengupta R, Ray SS, Bhattacharjee A, Ghosh S. Novel cellulases from an extremophilic filamentous fungi Penicillium citrinum: production and characterization. J. ind. Microbial. Biotechnol. 2008; 35:275-282.
Gupta R, Gigras P, Mohepata H, Goswani VK, Chuahan B. Microbial Amylases: A biotechnological perspective. Process Biochemistry 2003; 38 (11): 1599-1616.
Ali S, Sayed A, Sarker RI, Alam R. Factors Affecting Cellulose Production by Aspergillus terreus using water hyacinth. World Journal of Microbiology and Biotechnology. 1991; 7 (1): 62-66.
Doppelbauer R, Esterbauer H, Steiner W, Lafferty RM, Steinmuller H. The use of lignocellulosic wastes for production of cellulase by Trichoderma reesei. Appl. Environ. Microbiol. 1987; 26(5): 485-494.
Krishna C. Production of bacterial cellulases by solid state bioprocessing of banana wastes. Bioresour. Technol 1999; 69(3), 231-239.
Murao S, Sakamoto R, Arai M. Cellulase of Aspergillus aculeatus. In: Methods in Enzymology, Wood, W.A. and S.T. Kellog (Eds.). Vol. 160, Academic Press Inc., London. 1988;275-284.
Jecu L. Solid State Fermentation of agricultural wastes for endoglucanase production. 1999;11:1-5
Sarkar N, Aikat K. Aspergillus fumigatus NITDGPKA3 Provides for Increased Cellulase Production. International Journal of Chemical Engineering Volume 2014, Article ID 959845, 9 pages http://dx.doi.org/10.1155/2014/959845
Amaeze NJ, Okoliegbe IN, Francis ME. Cellulase production by Aspergillus niger and Saccharomyces cerevisiae using fruit wastes as substrates.International Journal of Applied Microbiology and Biotechnology Research. 2015;3: 36-44.
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