Integrity Research Journals

ISSN: 2536-7064
Model: Open Access/Peer Reviewed
DOI: 10.31248/JBBD
Start Year: 2016

Biochemical properties of α-amylase isolated from Aspergillus tamarii from cassava starch using solid state fermentation   |   Article Number: 9CB3731F6   |   Vol.8 (2) - April 2023

Received Date: 26 March 2023   |   Accepted Date: 29 April 2023  |   Published Date: 30 April 2023

Authors:  Opeyemi Olumuyiwa Otuewu1* , Sherifat Tolulope Akindele1, , Solomon Aderemi Akapo1 , Abdulrasak Alao Abdulkareem1 , Temitayo Adewale Adefuwa2 and Moronke Muinat Adeyanju2

Keywords: α-Amylase, Aspergillus tamari, cassava flour, ion-exchange chromatography, sweeteners.

Cassava starch can be completely hydrolyzed to produce a sweetener that can be used as a substitute for refined sugars and artificial sweeteners in the food and pharmaceutical industries. This work was designed to isolate and identify a good amylase producing fungus from cassava flour and to extract, purify, and partially characterize α-amylase produced. The enzyme was produced through solid-state fermentation followed by 70% ammonium sulphate precipitation and ion-exchange chromatography on Carboxyl-Methyl (CM) Sephadex C25. The physicochemical properties of the purified enzyme were determined. The peak with the highest activity was pooled from the latter chromatographic step and characterized afterward. The enzyme’s specific activity rose from 0.11 to 2.1 U/mg having a yield of 15.8% and a purification fold of 19.1. The optimal pH and temperature of the enzyme were 6.0 and 50°C respectively. The enzyme was observed to be thermo-stable at 50°C for 15 to 30 minutes. The kinetics revealed that the Vmax was 1.25 U/min while Km was 0.2 mg/ml. The enzyme’s native and sub-unit molecular weights were found to be 22 and 18.5KDa respectively. The results revealed conclusively that the isolated enzyme from Aspergillus tamarii exhibited the properties of glucoamylase.

Adeyanju, M. M., Ojewunmi, O., Akande, T., Ezima, E. N., Akeredolu, A. A., & Fagbohunka, B. S. (2014). Partial purification and some physicochemical properties of Aspergillus flavus α-amylase isolated from decomposing cassava peels. African Journal of Biotechnology, 13(52), 4657-4662.
Aisien, E. T., & Igbinosa, I. H. (2019). Production, purification, and characterization of α-amylase from Aspergillus niger, Aspergillus flavus and Penicillium expansum using cassava peels as substrate. Nigerian Journal of Biotechnology, 36(2), 114-126.
Akhter, P., Mishra, A., Mishra, V., & Raghav, A. (2017). Alpha amylase production from Aspergillus terreus UF39 using oil cakes, European Journal of Biotechnology and Bioscience, 5(6) 15-,21.
Aneja, K. R. (1996). Experiments in microbiology, plant pathology, tissue culture & mush room cultivation (2nd edition). Wishwa Prakashan New Delhi. p. 451.
Ashwini, K., Gaurav, K., Karthik, L., & Bhaskara Rao, K. V. (2011). Optimization, production and partial purification of extracellular α-amylase from Bacillus sp. marini. Archives of Applied Science Research, 3(1), 33-42.
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.
Aullybux, A. A., & Puchooa, D. (2013). α-Amylase production on low-cost substrates by Naxibacter sp. isolated from Mauritian soils. British Microbiology Research Journal, 3(4), 478-491.
Bagheri, A., Khodarahmi, R., & Mostafaie, A. (2014). Purification and biochemical characterisation of glucoamylase from a newly isolated Aspergillus niger: Relation to starch processing. Food Chemistry, 161, 270-278.
Bernfeld, P. (1955). Amylase, α and β. Methods in Enzymology, 1, 149-158.
Bradford, M. M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical biochemistry, 72(1-2), 248-254.
Chay, C., Dizon, E. I., Elegado, F. B., Norng, C., Hurtada, W. A., & Raymundo, L. C. (2017). Isolation and identification of mold and yeast in medombae, a rice wine starter culture from Kompong Cham Province, Cambodia. Food Research, 1(6), 213-220.
de Souza, P. M., & Magalhães, P. D. O. (2010). Application of microbial α-amylase in industry-A review. Brazilian Journal of Microbiology, 41, 850-861.
Deepika, M., & Satyanarayana, T. (2016). Bacterial and archaeal α-amylases: Diversity and amelioration of the desirable characteristics for industrial applications. Frontiers in Microbiology, 7, Article number 1129.
Hwang, K., Shin, S. G., Kim, J., & Hwang, S. (2008). Methano-genic profiles by denaturing gradient gel electrophoresis using order-specific primers in anaerobic sludge digestion. Applied Microbiology and Biotechnology, 80, 269-276.
Karlström, A., Calle, F., Salazar, S., Morante, N., Dufour, D., & Ceballos, H. (2016). Biological implications in cassava for the production of amylose-free starch: impact on root yield and related traits. Frontiers in plant science, 7, Article number 604.
Kayode, O. (2016). Exploring the cassava industry's value chain opportunities for MSMEs. Retrieved from https://www.linkedin. com/pulse/exploring-cassava-industrys-value-chain-msmes-kayode-omosebi.
Koç, Ö., & Metin, K. (2010). Purification and characterization of a thermostable glucoamylase produced by Aspergillus flavus HBF34. African Journal of Biotechnology, 9(23), 3414-3424.
Konsoula, Z., & Liakopoulou-Kyriakides, M. (2007). Co-production of α-amylase and β-galactosidase by Bacillus subtilis in complex organic substrates. Bioresource Technology, 98(1), 150-157.
Mesfin, M., Negussie, F. B., Mesfin, M., Manikandan, M., Melese, A. (2019). Isolation and characterization of amylase enzyme from selected fungal strains of Wof Washa Forest of North Shoa, Ethiopia. Science Journal of Biology & Life Science, 1(2): 1-10.
Metin, K., Koc, O., Ateşlier, B. B., & Biyik, H. H. (2010). Purification and characterization of α-amylase produced by Penicillium citrinum HBF62. African Journal of Biotechnology, 9(45), 7692-7701.
Moreira, F. G., Lenartovicz, V., de Souza, C. G., Ramos, E. P., & Peralta, R. M. (2001). The use of alpha-methyl-D-glucoside, a synthetic analogue of maltose, as inducer of amylase by Aspergillus sp in solid-state and submerged fermentations. Brazilian Journal of Microbiology, 32, 15-19.
Moreira, F. G., Lima, F. A. D., Pedrinho, S. R. F., Lenartovicz, V., Souza, C. G. M. D., & Peralta, R. M. (1999). Production of amylases by Aspergillus tamarii. Revista de Microbiologia, 30, 157-162.
Oboh, G. (2005). Isolation and characterization of amylase from fermented cassava (Manihot esculenta Crantz) wastewater. African Journal of Biotechnology, 4(10), 1117-1123.
Ominyi, M. C., Ogbonna, J. C., Nwoba, E. G., Nwagu, K. E., & Ukachi, R. (2013). Isolation, and screening of α-amylase and Glucoamylase producing fungi and their Glucomaylase producing fungi and their application in Bioethanol, production. International Journal Science Nature, 4(1), 44-50.
Saranraj, P., & Stella, D. (2013). Fungal amylase: A review. International Journal of Microbiological Research, 4(2), 203-211.
Schallmey, M., Singh, A., & Ward, O. P. (2004). Developments in the use of Bacillus species for industrial production. Canadian Journal of Microbiology, 50(1), 1-17.
Shah, I. J., Gami, P. N., Shukla, R. M., & Acharya, D. K. (2014). Optimization for α-amylase production by Aspergillus oryzae using submerged fermentation technology. Basic Research Journal of Microbiology, 1(4), 1-10.
Siddique, F., Hussain, I., Mahmood, M. S., Ahmed, S. I., & Iqbal, A. (2014). Isolation and characterization of a highly thermostable alpha-amylase enzyme produced by Bacillus licheniformis. Pakistan Journal of Agricultural Sciences, 51(2), 309-314.
Sidkey, N. M., Abo-Shadi, M. A., Balahmar, R., Sabry, R., & Badrany, G. (2011). Purification and characterization of α-amylase from a newly isolated Aspergillus flavus F2Mbb. International Research Journal of Microbiology, 2(3), 96-103.
Weber, K., & Osborn, M. (1975). Proteins and sodium dodecyl Sulphate: Molecular weight determination on polyacrylamide gels and related procedures. In: Hill, R. L.(ed.). The proteins (vol 1). New York, Academic Press. Pp. 179-223.