Molecular Screening and Production of α-amylase from Fungal sp.
DOI:
https://doi.org/10.14738/aivp.95.11036Keywords:
18s rDNA, α-amylase, fermentation, Optimization, Aspergillus tamarii T5Abstract
α-amylase producing Aspergillus sp. was isolated from decaying bread collected from bakery site located within Enugu metropolis, Enugu state. Standard microbiology, biochemical and molecular techniques (18s DNA sequencing) were used for confirmation of the fungal strain as Aspergillus tamari.. p-NPG infused nutrient broth confirmed Aspergillus tamarrii as prolific producer of α-amylase with rapid yellow colouration after forty eight hours of incubation. Solid state fermentation on rice bran matrix (SSF) system was used for the enzyme production. Cork borer of 2mm diameter was used throughout the production and optimization studies. Carbon sources including: Starch, wheatbran and sugarcane baggase were optimized, starch was found suitable for the protein production with highest α-amylase activity (91.71 μmol/min). Among the nitrogen sources optimized, peptone was found optimal for α-amylase production with activity of 90.34 μmol/min. pH 6.0 was found the best for the enzyme production. Effect of incubation period on the enzyme production showed the 4th day of fermentation as the peak day for α-amylase production from Aspergillus tamari. The results from this study have shown that Aspergillus tamari among other fungal isolates from mould bread collected from bakery sites in Enugu metropolis has the potential for α-amylase production in a commercial scale for both industrial and clinical applications..
References
Adefila, O., Bakare, M. and Adewale, O. (2012).Characterization of α-amylase from sorghum (sorghum bicolor) obtained under optimized conditions. Journal ofInstitute of Brewing,118(1):63-69
Aiyer, P.V. (2004). Effect of C:N ratio on α-amylase production by Bacillus licheniformis SPT 278.AfricanJournal of Biotechnology,3: 519–522.
Allam, R., Aly, M., El-zhrany, K. and Shafei, M. (2016).Production of β-Galactosidase Enzyme fromLactobacillus acidophilus RK isolated from different sources of milk and dairy products. International Journal of ChemTech Research, 9 (10): 218-231.
Amritkar, N., Kamat, M. and Lali, A.(2004).Expanded bed affinity purificationof bacterial a-amylase and cellulase on composite substrate analogue–cellulose matrices. Process Biochemistry,39:565–570.
Babu, K. and Satyanarayana, T. (1995)α-amylase production by thermophilic Bacillus coagulans in solid-state fermentation.ProcessBiochemistry,30: 305–309
Balkan, B. and Ertan, F. (2005).Production and properties of α-amylase from Penicillium chrysogenum and its application in starch hydrolysis. Preparative Biochemistry and Biotechnology,35, 169-178.
Barnett, H. and Hunters B. (1972).Illustration Genera of Imperfecti Fungi.3rd edn. Burgess Publishing Company, London. pp.273-274.
Baysal, F., Uyar, F. and Aytekin, C. (2003).Solid-state fermentation for production of a-amylase by a thermotolerant Bacillus subtilis from hot-spring water. Process Biochemistry,38:1665–1668.
Bessler, J.,Schmitt, K., Maurer, D. and Schmid, D. (2003): Directedevolution of a bacterial a-amylase: Toward enhanced pHperformance and higher specific activity. Protein Science,12:2141–2149.
Burhan, A., Nisa, U., Gokhan, C., Omer, C., Ashabil, A. and Osman, S. (2003). Enzymatic properties of a novel thermostable, thermophilic, alkaline and chelator resistant amylase from analkaliphilic Bacillus sp. isolate ANT-6.ProcessBiochemistry,38: 1397–1403.
Carlos Alberto, M., Meire, L. and Luciano, A. (2002). Production of α-Amylase from thermophilic Bacillus sp. Brazillian Journal of Microbiology, 33(1): 1517-8382.
Chen, B., Li, D. and Yang S. (2005).Isolation of acid α-amylase producing strain and purification and characterization of acid α-amylase.Food Science, 26: 119-122.
Chinny, R. and Diamond, R. (1982).Generation of chemotactic factors by Rhizopus oryzae in the presence and absence of serum: relationship to hyphal damage mediated by human neutrophils and effects of hyperglycemia and ketoacidosis. Infection and Immunity,38 (3): 1123–29.
Cohn, M. and Monod, J. (1951). Purification etproprietes de la β-galactosidase (lactase) d'Echerichia coli. BiochimBiophysActa.7:153–174.
Corbishley, D. and Miller,W. (1984). Tapioca, arrow Root and Sago Starches Production Production. In Starch Chemistry and Technology, Whistler, R., Be Miller, J and Paschal, E. Academic Press, New York.pp 469-476.
Dakhmouche, S., Gheribi-Aoulmi, Z., Meraihi, Z. and Bennamoun, L. (2005). Application of a statistical design to the optimizationof culture medium for a-amylase production byAspergillus nigerATCC 16404 grown on orange waste powder. Journal of Food Engineering,73: 190–197.
Declerck, N., Machius, M., Joyet, P., Wiegand, G., Huber, R. and Gaillardin, C. (2003) Hyperthermostabilisation of Bacillus licheniformisa-amylase and modulation of its stability over a 50°C temperature range. Protein Engineering,16: 287–293.
Ezeonu, M., Okafor, J., Ogbonna, J. (2013).Laboratory Exercises in Microbiology.1st edn. Ephrata Publishing and Printing Company, Nsukka. Pp 100-117. st
Ezeonu, M., Okafor, J., Ogbonna, J. (2013).Laboratory Exercises in Microbiology. 1 Edn. Ephrata Publishing and Printing Company, Nsukka. pp 100-117.
Gheytanchi, E.,Heshmati, F.,Kordestani, B., Nowroozi, J. and Movahedzadeh, F. (2010). Study on β-galactosidase enzyme produced by isolated Lactobacilli from milk and cheese. African Journal of Microbiology Research, 4(6), 454-458.
Goyal, N., Gupta, K., and Soni, S. (2005 ). A novel raw starch digesting thermostable α-amylase from Bacillus sp. I-3 and its use in the direct hydrolysis of raw potato starch. Enzyme and Microbial Technology, 37(7):723–734.
Khajeh, K. and Nemat-Gorgani, M. (2001).Comparative studies on amesophilic and a thermophilic α-amylase.AppliedBiochemicalBiotechnology,90: 47–55.
Kumar, P., Nampoothiri, M., Ramachandran, S., Szacks, G. and Pandey, A. (2004).Partial purification and characterization of α-amylase produced from Aspergillus oryzae Kumar, S., Stecher, G. and Tamura, K. 92016).Molecular evolutionary genetics analysis version 7.0 for bigger datasets.Molecular Biology and Evolution, 33(7): 1870- 1874.
Kunamneni, A. and Singh, S.(2005) Response surface optimization ofenzymatic hydrolysis of maize starch for higher glucose production.BiochemicalEngineeringJournal,27:179–190.
Lowry, O.,Roseburg,N.,Farr,A. and Randall,R .(1951).Protein Measurement with Folin- Phenol Reagents. Journal of Biological Chemistry, 93:265-275.
MacGregor, E. A. (2005).An overview of clan GH-H and distantly-related families.Biologia (Bratislava), 60: 5–12.
Agwu, C. H. (2021). Molecular Screening and Production of α-amylase from Fungal sp. European Journal of Applied Sciences, 9(5). 365-379.
Miller, G. (1959). Use of Dinitrosalicyclis Acid Reagent for Reducing Sugar.AnalyticalChemistry,31:426-442.
Mishra, S., Noronha, S. and Suraishkumar, G. (2005). Increase inenzyme productivity by induced oxidative stress in Bacillus subtilis cultures and analysis of its mechanism using microarray data, Process Biochemistry,40: 1863–1870.
Mitsuiki, S., Mukae, K., Sakai, M., Goto, M., Hayashida, S. and Furukawa, K.(2005).Comparative characterization of raw starch hydrolyzing α-amylases from various Bacillus strains.Enzyme andMicrobialTechnology,37: 410–416.
Mohamed, L., Zakaria, M., Ali, A.; Senhaji, W.; Mohamed, O., Mohamed, E., Hassan, B. and Mohamed, J. (2007). Optimization of Growth and Extracellular Glucoamylase Production by Candida famata Isolate. African Journal of Biotechnology,6: 2590-2595.
Nwagu, T., and Okolo, N. (2011). Extracellular amylase production of a thermo tolerant Fusariumspecies isolated from Eastern Nigerian soil. Brazilian Archive of Biology andTechnology, 54(4): 1516-8913.
Okolo, B., Ezeogu, L. and Mba, C. (1995). Production of raw starchdigesting amylase by Aspergillus nigergrown on native starch sources.JournalofScienceand Food Agriculture,69:109–115.
Shen, H., Mo, X., Chen, X., Han, D. and Zhao, C. (2012).Purification and enzymatic identification of an acid stable and thermostable α-amylase from Rhizopus microsporus.The Institute of Brewing and Distilling, 118: 309-314.
Silva, T., Attilis Angelis, D., Carvalho, A., Silva, R., Boscolo, M. and Gomes E. (2005).Production of saccharogenic and dextrinogenic amylases by Rhizomucorpussillus.Journal of Microbiology, 43(6)561-568.
Singh, R., Kumar, V. and Kapoor, V. (2014). Partial purification and characterization of a heat stable α-amylase of thermophilicActinobacter, Streptomyces sp.MSC 702. Enzyme Research, 1-8.
Sudha. (2013). Effects of different concentrations of metal ions on alpha amylase produced by Baccilusamyloliquefacians. Research in Biotechnology, 3(4): 67-71.
Vander Maarel,M., Vanderveen, B., Uitdehaag, J., Leemhurs, H., and Dijkhuizen, L. (2002).Properties and application of starch converting enzyme of alpha amylase Family. Journal of Biotechnology, 94:137-155.
Vieille, K. and Zeikus, G. (2001). Hyperthermophilic enzymes: Sources,uses, and molecular mechanisms of thermostability. Microbiology and MolecularBiologyRevised,65:1–43.
Vijayaraghavan, P., Remy, C. and Prakash, V. (2011).Production of α-amylase by Rhizopus microsporus using agricultural by-products in solid state fermentation. Research Journal of Microbiology, 6: 366-375.
Downloads
Published
How to Cite
Issue
Section
License
Copyright (c) 2021 Cletus H. Agwu
This work is licensed under a Creative Commons Attribution 4.0 International License.
