Isolation, Screening and Time Course Study of Amylase-producing Fungi from Garri Processing Environment


Garri Processing
Time Course

How to Cite

Ire, F., Wokoma, S. I., & Okoli, A. (2020). Isolation, Screening and Time Course Study of Amylase-producing Fungi from Garri Processing Environment. Journal of Life and Bio Sciences Research , 1(3), 68 - 75.


Amylases (E.C. are enzymes which catalyze the breakdown of -1,4 glycosidic linkages of starch to simple sugars and different monomeric products. Microbial amylases are essential enzymes compared with animals and plants amylases in industrial applications. Thus, in southern Nigeria, garri processing waste-water from mills poses a serious environmental challenge and this situation could be exploited by utilizing microorganisms colonizing the area to manufacture microbial products. This present study was aimed at evaluating the potential of fungi isolated from garri processing environment for amylase secretion and conduct a time course study of the enzyme production. Fungi were isolated from the soil and waste water from garri processing sites using Standard Microbiological Procedures. The five fungi isolated from garri processing environmental samples were screened using Lugols iodine. Three fungal isolates were then selected on the hydrolysis of starch in qualitative zonation agar plates but one isolate named B among them was better in starch hydrolysis related to highest clear zone plates. After qualitative screening, the three hyper production amylase extracted fungi were identified according to the morphological characteristics. The hyper producing amylase isolate B was then identified as Aspergillus flavus (RCBBR_AEAFUN2) and recorded as a novel strain in southern Nigeria according to molecular characteristics, which was selected for further studies. The current results of time course study showed that the maximum growth (2.453 g) occurred after 24 h of incubation at pH 4.5 while maximum enzyme production (2.3 U/ml/min) was obtained after 96 h of incubation at pH 3.5. Therefore, the hyper producing amylase from garri processing sites, Aspergillus flavus (RCBBR_AEAFUN2) novel strain has great prospects for future biotechnological and industrial applications and help in ameliorating the environmental pollution posed by the waste-water.


Aiyer, P.V. (2005). Amylases and their applications. African Journal of Biotechnology, 4(13), 1525-1529.

Asrat, B., Girma, A. (2018). Isolation, production and characterization of amylase enzyme using the isolate Aspergillus niger FAB-211. International Journal of Biotechnology and Molecular Biology Research, 9(2), 7-14.

Ayansina, A.D.V., Adebola, M.A., Adeyemi, A.O. (2014). Some microorganisms associated with soils exposed to cassava (Mannihot esculatum) peels. American Journal of Research Comm eunication, 2(9), 155-162.

Bakri, Y., Magali, M., Thonart, P. (2009). Isolation and identification of a new fungal strain for amylase biosynthesis. Polish Journal of Microbiology, 58(3), 269-273.

Batista, E., Watanabe, J.Y.M., Oliveira, V.M., Passarini, M.R.Z., (2018). Avalia羡o da produ羡o de amilase e protease por bacterias da Antartica. Revista Brasileira de Inicia羡o Cientí¦©ca, 5, 1329.

Cappuccino, J.G., Sherman, N. (2004). Microbiology a Laboratory Manual, (6th edition) Pearson Education, Pvt. Ltd, Delhi, India.

Dash, B.K, Rahman, M.M., Sarker, P.K. (2015). Molecular identification of a newly isolated Bacillus subtilis BI19 and optimization of production conditions for enhanced production of extracellular amylase. BioMed Research International, 9.

Elijah, A.I., Asamudo, N.U. (2016). Molecular characterization and potential of fungal species associated with cassava waste. British Biotechnology Journal, 10(4), 1-15.

Fadahunsi, I.F., Garuba, O.E. (2012). Amylase production by Aspergillus flavus associated with the bio-deterioration of starch-based fermented foods. New York Science Journal, 5(1), 13-18.

Fang, W, Xue, S., Deng, P., Zhang, X., Wang, X., Xiao, Y., Fang, Z. (2019). AmyZ1: a novel -amylase from marine bacterium Pontibacillus sp. ZY with high activity toward raw starches. Biotechnology for Biofuels, 12:95.

Felsenstein, J. (1985). Confidence limits on phylogenies: An approach using the Bootstrap. Society for the Study of Evolution, 39, 783-791.

Gautam, N., Singh, K.P., Modi, D.R. (2013). Screening of soil fungi for -amylase activity. Journal of Recent Advances in Applied Sciences, 28, 110-112.

Geetha, K.N., Jeyaprakash, K., Nagaraja, Y.P. (2011). Isolation, screening of Aspergillus flavus and its production parameters for amylase under solid state fermentation. Journal of Applied and Natural Science, 3(2), 268-273.

Gopinath, S.C.B., Anbu, P., Arshad, M.K.M, Lakshmipriya, T., Voon, C.H., Hashim, U., Chinni, S.V. (2017). Hindawi BioMed Research International, 9 Pages,

Gupta, A., Gupta, V.K., Modi, D.R., Yadava, L.P. (2008). Production and characterization of -amylase from Aspergillus niger. Biotechnology, 7(3), 551-556.

Ibraheem, L.J., Mezeal, I.A., Mizil, S.N. (2017). Manufacture enzyme from the fungus A. flavus. World Journal of Pharmaceutical Research, 6(10), 117-125.

Ire, F.S., Ike, V.C. (2014). Screening and Optimization of Process Parameters for the Production of Lipase in Submerged Fermentation by Aspergillus carbonarius (Bainer) IMI 366159. Annual Research & Review in Biology, 4(16), 2587-2602

Ire, F.S., O.C. Eruteya, O.C., Amaechi, V. (2017). Optimization of culture conditions using One-Factor-at-Time Methodology and partial purification of amylase from Aspergillus niger of DTO: H5 under solid state fermentation. International Journal of Current Microbiology and Applied Sciences, 6(5): 307-325.

Ire, F.S., Nwanguma, A.C. (2020). Comparative Evaluation on Tannase Production by Lasiodiplodia plurivora ACN-10 under Submerged Fermentation (SmF) and Solid State Fermentation (SSF). Asian Journal of Biotechnology and Bioresource Technology, 6(1), 39-49,

Jukes, T.H., Cantor, C.R. (1969). Evolution of protein molecules. Mammalian protein metabolism, Academic press, New York, 3, 21-132.

Kolawole, O.P. (2014). Cassava processing and the environmental effect. World Sustainability Forum 2014-Conference Proceedings Paper.

Lonsane, B.K., Ghildyal, N.P., Budiatman S, Ramakrishna, S.V. (1985) Engineering aspects of solid-state fermentation. Enzyme Microbial Technology,7(6), 258-265.

Lowry, O.H., Rosebrough, N.J., Farr, A.L., Randall, R.J., (1951). Protein measurement with the Folin-Phenol reagents. Journal of Biological Chemistry, 48, 17-25.

Miller, G.L. (1959). Use of dinitrosalicylic acid reagent for determination of reducing sugar. Analytical Chemistry, 31(3), 426428.

Negussie F Bussa, Mesfin Moges, Manikandan Muthuswamy, Melese Abdisa (2019). Isolation and characterization of amylase enzyme from selected fungal strains of Washa Forest of North Shoa, Ethiopia. Science Journal of Biology & Life Sciences. 1(2). SJBLS.MS.ID.000506.

Oboh, G. (2005). Isolation and characterization of amylase from fermented cassava (Manihot esculenta Crantz) waste-water. African Journal of Biotechnology, 4 (10), 1117-1123.

Obueh, H. O., Odesiri-Eruteyan, E. (2016). A study on the effects of cassava processing wastes on the soil environment of a local cassava mill. Journal of Pollution Effects and Control, 4(4), 1-4 doi:10.4176/2375-4397.1000177

Ogbonna, C. N., Okpokwu, N. M., Okafor, C. U., Onyia, C. E. (2014). Isolation and screening of amylase producing fungi obtained from garri processing site. International Journal of Biotechnology and Food Science, 2 (5), 88-93.

Okolo, B.N., Ire, F.S, Ezeogu, L.I., Anyanwu, C.U., Odibo, F.J.C. (2000). Purification and some properties of a novel raw starch digesting amylase from Aspergillus carbonarius. Journal of the Science of Food and Agriculture, 81, 329 336.

Omilani, O., Abass, A.B., Okoruwa, V.O. (2019). Smallholder agroprocessors willingness to pay for value-added solid waste management solutions. Sustainability, 11, 1759, doi:10.3390/su11061759

Ominyi, M.C., Ogbonna, J.C., Nwoba, E.G., Nwagwu K.E. Ukachi, R. (2013). Isolation and Screening of a-amylase and Glucoamylase Producing Fungi and their Application in Bioethanol Production. International Journal of Science and Nature, 4(1), 44-50.

Ottoni, J.R., de Silva, T.R., de Oliveira, V.M., Passarini, M.R.Z. (2020). Characterization of amylase produced by cold-adapted bacteria from Antarctic samples. Biocatalysis and Agricultural Biotechnology, 23 (2020) 101452, 1-8.

Pham, J.V., Yilma, M.A., Feliz, A., Majid, M.T., Maffetone, N., Walker, J.R., Kim, E., Cho, H.J., Song, M.C., Park, S.R., Yoon, Y.J. (2019). A review of the microbial production of bioactive natural products and biologics. Frontiers in Microbiology, 2019,

Pruitt, K.D., Tatusova, T., Maglott, D.R. (2005). NCBI Reference Sequence (Ref- Seq): A Curated Non-Redundant Sequence Database of Genomes, Transcripts and Proteins. Nucleic Acids Research, 33:501-504.

Saghai-Maroof, M.A., Soliman, K.M., Jorgensen, R.A., Allard, R.W. (1984). Ribosomal DNA spacer-length polymorphisms in barley: Mendelian inheritance chromosomal location and population dynamics. Proceedings of the National Academy of Sciences of the United States of America, 81, 8014-8018.

Saini, R., Saini, H.S., Dahiya, A. (2017). Amylases: characteristics and industrial applications. Journal of Pharmacognosy and Phytochemistry, 6(4), 1865-1871.

Saitou, N., Nei, M. (1987). The neighbor-joining method: A new method for reconstructing phylogenetic trees. Molecular biology and evolution, 4, 406-425.

Shafique, S., Bajwa, R., Shafique, S. (2009). Screening of Aspergillus niger and A. flavus strains for extracellular alpha-amylase activity. Pakistan Journal of Botany, 41(2): 897-905.

Singh, R., Mittal, A., Kumar, M., Mehta, P.K. (2016). Amylases: A note on current applications. International Research Journal of Biological Sciences, 5(11): 27-32

Sunitha, V.H., Ramesha, A., Savithas, C. (2012). Amylase production by endophytic fungi Cylindrocephalum sp. isolated from medicinal plant Alpinia calcarata (Haw.). ROSCOE. Brazilian Journal of Microbiology, 1213-1221.

Tamura, K., Dudley, J., Nei, M., Kumar, S. (2007). MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) Mol. Bio. Evol., 24:1596-1599.

Creative Commons License

This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License.

Copyright (c) 2020 Array