Production of Glucoamylase, α-amylase and Cellulase by Aspergillus oryzae F-923 Cultivated on Wheat Bran under Solid State Fermentation

Main Article Content

M. Fadel
Sawsan AbdEl-Halim
Hayat Sharada
Ahmed Yehia
Mayar Ammar


Filamentous fungi have been widely utilized in production of enzymes which have many industrial applications. In this study, twenty five local fungal isolates, belonging to Aspergillus sp., Trichoderma sp. and Penicillium sp., were screened under solid state fermentation conditions (SSF) for the production of α-amylase, glucoamylase and cellulase. Asperigillus oryzae F-923, cultivated on wheat bran, was the most promising isolate for production of the target enzymes under this study. Physical parameters of moisture content, pH, temperature and incubation time, optimized were 1:2(w/v), 5.5, 28°C and 72 hr, respectively. The production of enzymes was enhanced when ammonium sulfate was supplemented as a nitrogen source to wheat bran. The production of α-amylase and glucoamylase was also enhanced when 10% (w/w) soluble starch was added as a carbon source to wheat bran. However xylose supplementation at 10% (w/w) was observed to be best for cellulase production. Tap water was found to be efficient for enzymes' extraction from the fermentation medium. Three successive extractions were needed to obtain the produced enzymes from the fermented substrate. Characterization of the produced enzymes revealed that, the optimum temperature for α-amylase and glucoamylase was 60°C, while 50°C was the optimum temperature for cellulase activity. Isopropanol 1:1(v/v) was proved to be more suitable for partial purification of enzymes. Following partial purification of enzymes of glucoamylase, α-amylase and cellulase increased to 10.8, 11.8 and 11.4 folds, respectively.

Glucoamylase, α-amylase, cellulose, Aspergillus oryzae, solid state fermentation.

Article Details

How to Cite
Fadel, M., AbdEl-Halim, S., Sharada, H., Yehia, A., & Ammar, M. (2020). Production of Glucoamylase, α-amylase and Cellulase by Aspergillus oryzae F-923 Cultivated on Wheat Bran under Solid State Fermentation. Journal of Advances in Biology & Biotechnology, 23(4), 8-22.
Original Research Article


Ramachandran S, Patel KA, Nampoothiri KM, Francis F, Nagy V, Szakacs G, Pandey A. Coconut oil cake a potential raw material for the production of α-amylase. Bioresource Technology. 2004;93:169-174.

Pandey A. Recent developments in solid-state fermentation. Process Biochemistry. 1992;27:109-117.

Raimbault M. General and microbiological aspects of solid substrate fermentation. Electronic Journal of Biotechnology. 1998;1(3):174-188.

Thomas L, Larroche C, Pandey A. Current developments in solid-state fermentation. Biochemical Engineering Journal. 2013;81: 146-161.

Raveendran S, Parameswaran B, Ummalyma SB, Abraham S, Mathew AK, Madhavan A, Rebello S, Pandey A. Applications of microbial enzymes in food industry. Food Technology and Biotechnology. 2018;56(1):16–30.

Subramaniyam R, Vimala R. Solid state and submerged fermentation for the production of bioactive substances: A comparative study. Journal of Science and Nature. 2012;3(3):480-486.

Keera AA, Mouafi FE, Kahil T, Fadel M, Abedo A. Hyper production of glucoamylase by Aspergillus oryzae FK-923 under solid state fermentation. World Applied Sciences Journal. 2014;30(11): 1447-1461.

Bhargav S, Panda BP, Ali M, Javedb S. Solid-state fermentation: An overview. Chem. Biochem. Eng. Q. 2008;22(1):49–70.

Karri S, Talla SG, Renuka S. Screening and production optimization of alpha amylase from Aspergillus strains by using solid state ferment. Int. J. Curr. Microbiol. App. Science. 2014;3(4):623-631.

Imran M, Anwar Z, Irshad M, Asad MJ, Ashfaq H. Cellulase production from species of fungi and bacteria from agricultural wastes and its utilization in industry: A review. Advances in Enzyme Research. 2016;4:44-55.

Haq I, Hameed U, Mahmood Z, Javsd MM. Solid state fermentation for the production of alpha amylase by Paenibacillus amylolyticus. Pak. J. Botony. 2012;44:341-346.

Anto H, Trivedi UB, Patel KC. Glucoamylase production by solid-state fermentation using rice flake manufacturing waste products as substrate. Bioresoure Technology. 2006;97(10):1161–1166.

Puri S, Arora M, Sarao L. Production and optimization of amylase and glucoamylase using Aspergillus oryzae under solid state fermentation. International Journal of Research in Pure and Applied Microbiology. 2013;3(3):83-88.

Kottwitz B, Schambil F. Cellulase and cellulose containing detergent. US 20050020472 A1.

Chen Y, Wan J, Zhang X, Ma Y, Wang Y. Effect of heating on recycled properties of unbleached eucalyptus cellulose fiber. Carbohydrate Polymers. 2012;87:730- 736.

Fadel M. Production of therrnostable amylolytic enzymes by Aspergilltus niger F-909 under solid state fermentation. Egypt. J. Microbiology. 2000;35(4):487-505.

Zafar S. Biofuels from lignocellulosic biomass, bio energy consult mass resource. 2020;5:20-35.

Zafar S. Ethanol from lignocellulosic wastes. Bioenergy Consult Mass Resource. 2019;10:13.27.

Lakshmi MVC, Jyothi P. Production and optimization of glucoamylase from Aspergillus oryzae NCIM 1212 using wheat bran, varying chemical parameters under solid state fermentation. Int. J. Curr. Microbiol. App. Science. 2014;3(5):70-76.

Smith RW, Misra M, Dubel J. Mineral bioprocessing and future minerals Engineering. 1991;4:1127-114.

Yehia A, Khalek MA, Ammar M. Cellulase as a new phosphate depressant in dolomite-phosphate flotation. Physicochem. Probl. Miner. Process. 2017;53(2):1092–1104.

Yehia A, Yassin KE, Ammar M. Upgrading of phosphate fines by fatty acid flotation using amylase enzyme as a surface modifier. Mining, Metallurgy & Exploration. 2019;36:949–955.

Hassan HM, Abeer A. Keera, Fadel M. High-yield cellulases and xylanase production from sugar-cane bagasse pith by Aspergillus oryzae FK-923 cultivated under solid state fermentation. Int. J. Curr. Res. Aca. Rev. 2016;4(1):1-13.

Miller GL. Use of dinitrosalicyclic acid reagent for determination of reducing sugar. Analytical Chemistry. 1959;31:426-429.

Ghose TK. Measurement of cellulase activities. Pure & Applied Chemistry. 1987;59(2):257-268.

Medda S, Chandra AK. New strains of Bacillus licheniformis and Bacillus coagulans producing thermostable α-amylase active at Alkaline pH. J. Appl. Microbiology. 1980;48(1):47-58.

Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Journal of Biological Chemistry. 1951;193:265-275.

Sajith S, Priji P, Sreedevi S, Benjamin S. An overview on fungal cellulases with an industrial perspective. J. Nutr. Food Sci. 2016;6(1):1-13.

Zambare V. Solid state fermentation of Aspergillus oryzae for glucoamylase production on agro residues. Int. J. Life Science. 2010;4:16-25.

Farid MA, Shata HMA. Amylase production from Aspergillus oryzae LS1 by solid-state fermentation and its use for the hydrolysis of wheat flour. Iranian Journal of Biotechnology. 2011;9(4):267-27.

Pandey A, Soccol CR, Mitchell D. New developments in solid state fermentation: I-Bioprocesses and products. Process Biochemistry. 2000;35(10):1153–1169.

Parbat R, Singhal B. Production of Glucoamylase by Aspergillus oryzae under solid state fermentation using agro-industrial products. Int. J. Microbiol. Res. 2011;2(3):204-207.

Kiran EU, Antoine P, Trzcinski Liu Y. Glucoamylase production from food waste by solid state fermentation and its evaluation in the hydrolysis of domestic food waste Biofuel Research Journal. 2014;3:98-105.

Abdalwahab SA, Ibrahim SA, Dawood ES. Culture condition for the production of glucoamylase enzyme by different isolates of Aspergillus spp. International Food Research Appl. Journal. 2012;19(3):1261-1266.

Ahmed MM, Soad A. El-Zayata, El-Sayed MA. Cellulolytic activity of cellulose-decomposing fungi isolated from Aswan hot desert soil, Egypt. Journal of Biological Studies JBS. 2018;1(2):35-48.

Bhargav S, Panda BP, Ali M, Javedb S. Solid-state fermentation: An overview. Chem. Biochem. Eng. Q. 2008;22(1):49–70.

Kumar B, Bhardwaj N, Alam A, Agrawal K, Prasad H, Verma P. Production, purification and characterization of an acid/alkali and thermo tolerant cellulase from Schizophyllum commune NAIMCC-F-03379 and its application in hydrolysis of lignocellulosic wastes. AMB Express. 2018;8(1):173.

Singhal B. Production of glucoamylase by Aspergillus oryzae under solid state fermentation using agro industrial products. International Journal of Microbiological Research. 2011;2(3):204-207.

Santos MM, Rosa AS, Dal’boit S, Mitchell DA, Kriger N. Thermal denaturation: Is solid state fermentation really a good technology for the production of enzymes? Bioresource Technology. 2004;93:261-268.

Goto CE, Barbosa EP, Kistner L, Do CL, Gandra KRF, Arrias VL, Peralta RM. Production of amylases by Aspergillus fumigatus. Revista de Microbiologia. 1998;29:99-103.

Abdalwahab SA, Ibrahim SA, Dawood ES. Culture condition for the production of glucoamylase enzyme by different isolates of Aspergillus spp. International Food Research Appl. Journal. 2012;19(3):1261-1266.

Bertolin TE, Schmidell W, Maiorano AE, Casara J, Costa JAV. Influence of carbon, nitrogen and phosphorous sources on glucoamylase production by Aspergillus awamori in solid state fermentation. Zeitschriftfür Naturforschung C: A Journal of Bioscience. 2003;58:708-712.

Kunamneni A, Permaul K, Singh S. Amylase production in solid state fermentation by the thermophilic fungus Thermomyces lanuginosus. Journal of Bioscience and Bioengineering. 2005;100: 168-171.

Shivaramakrishnam S, Ganghadharan D, Nampoothiri KM, Soccol CR, Pandey A. Alpha amylase production by Aspergillus oryzae employing solid-state fermentation. Journal of Scientific and Industrial Research. 2007;66:621-626.

Sher H, Abdul Ghani MF, Mehmood R, Rehman H, Bokhari SAL. Optimization of cellulase enzyme production from Aspergillus oryzae for industrial application. World J. Biology and Biotechnology. 2017;2(2):55-158.

Karim BMR, Tasnim T. Fungal glucoamylase production and characterization: A review. 2018;4(2):591-609.

Ezugwu AL, Eze SOO, Chilaka FC. A study of the optimal conditions for glucoamylase obtained from Aspergillus niger using amylopectin from cassava starch as carbon source. Afric. J. Biotech. 2015;14(36):2693-2702.

Silveira ED, Gomes E, Souza SR, Grandi RP. Production of thermostable glucoamylase by newly isolated Aspergillus flavus A.1.1 and Thermomyces anuginosus A 13.37. Braz. J. Microbiol. 2005;36:75-82.

Chandra MS, Viswanath B, Reddy RB. (Optimization of extraction of β-endoglucanase from the fermented bran of Aspergillus niger. Indian Journal of Microbiology. 2010;50:122-126.

Gao J, Weng H, Zhu D, Yuan M, Guan F, Xi Y. Production and characterization of cellulolytic enzymes from the thermoacidophilic fungal Aspergillus terreus M11 under solid-state cultivation of corn stover. Bioresource Technology. 2008;99(16):7623-7629.

Pirota RD, Delabona PS, Farinas CS. Simplification of the biomass to ethanol conversion process by using the whole medium of filamentous fungi cultivated under solid-state fermentation. Bioenergy Research. 2014;7:744– 752.

Mala JGS, Edwinoliver NG, Kamini NR, Puvanakrishnan R. Mixed substrate solid state fermentation for production and extraction of lipase from Aspergillus niger MTCC 2594. J. Gen. Appl. Microbiology. 2007;53:247-253.

Shata HMA. Extraction of milk-clotting enzyme produced by solid state fermentation of Aspergillus oryzae. Polish Journal of Microbiology. 2005;54(3):241-247.

Khodarahmi R, Mostafaie A. Purification and biochemical characterisation of glucoamylase from a newly isolated Aspergillus niger: Relation to starch processing. Food Chemistry. 2014;161: 270-27.

Lam WC, Pleissner D, Lin CSK. Production of fungal glucoamylase for glucose production from food waste biomolecules. 2013;3(3):651–661.

Riou C, Salmon JM, Vallier MJ, Günata Z, Barre P. Purification, characterization, and substrate specificity of a novel highly glucose-tolerant beta-glucosidase from Aspergillus oryzae. Appl Environ. Microbiology. 1998;64(10):3607-14.

Begum MF, Alimon AR. Assessment of some wild Aspergillus BS1. Ann Microbiol. 2011;64:763-771.

Prajapati BP, Suryawanshi RK, Agrawal S, Ghosg M, Kango N. Characterization of cellulase from Aspergillus tubingenesis NKBP-55 for generatin of fermentable sugars from agricultural residues. Bioresource Technology. 2018;250:733-740.