Advances in Biochemistry

| Peer-Reviewed |

Biochemical Characterization of Crude α-Amylase of Aspergillus spp. Associated with the Spoilage of Cassava (Manihot esculenta) Tubers and Processed Products in Nigeria

Received: 22 January 2015    Accepted: 01 February 2015    Published: 09 February 2015
Views:       Downloads:

Share This Article

Abstract

In this research, crude α-amylases associated with the spoilage of cassava (Manihot esculenta) tubers/ product (‘eba’) were biochemically characterized. They were isolated from five fungi: Aspergillus sp. CSA25, Aspergillus sp. CSA26, Aspergillus sp. CSA27, Aspergillus sp. CSA35 and Aspergillus sp. CSA38. The results of the analyses showed that the activities of α-amylase obtained from both sources (cassava tuber/ eba) were optimal at 45°C and pH 5.0. The maximum specific activity (Vmax) of the enzyme was found to be 10 U/mg protein, while its Michaelis-Menten constant (Km) was between 0.37 -1.25%w/v. The α-amylase is thermally stable for 1 - 2 h at optimum temperature and pH (45°C; pH 5.0). A broad range of substrate specificity was expressed by the enzyme for cassava starch-containing products (tapioca flour, garri flour, cassava flour, 1%, w/v); however, potato (Ipomoea batatas) starch, yam (Dioscorea rotundata) flour and cocoyam (Colocasia esculenta) flour were relatively minimally hydrolyzed by the crude α-amylases obtained from Aspergillus spp. that caused spoilage of cassava. Ethylenediamine tetraacetic acid (1 mM EDTA) and Mg2+ treatment had no significant (p > 0.05) effect on the activities of the amylase, but Na+, K+, Ca2+, Fe3+, thiourea and 5′,5′-dithiobis-2-nitrobenzoate (1 mM DTNB) enhanced its activities. The fungal α-amylases were most activated by K+ and had a salt tolerance of 1 - 2 M NaCl for 24 h. The fungal α-amylases reported in this study would find useful application in industries like food industry, detergent industry, paper industry, textile industry, pharmaceutical industry, etc where microbial α-amylases would be required for efficient and cost-effective hydrolysis of cassava starch, cassava flour and or its products.

DOI 10.11648/j.ab.20150301.14
Published in Advances in Biochemistry (Volume 3, Issue 1, February 2015)
Page(s) 15-23
Creative Commons

This is an Open Access article, distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution and reproduction in any medium or format, provided the original work is properly cited.

Copyright

Copyright © The Author(s), 2024. Published by Science Publishing Group

Keywords

Manihot esculenta (Cassava), Aspergillus spp., α-Amylase, Biochemical Characterization

References
[1] Abdullah, R., and Ikram-ul-Haq. Purification and characterisation of α-amylase produced by mutant strain of Aspergillus oryzae EMS-18. Natural product research (ahead-of-print), 2014; 1-7.
[2] Adeniran, H. A., and Abiose, S. H. Partial purification, characterization and hydrolytic activities of amylases from Bacillus licheniformis and Aspergillus niger cultured on agricultural residues. Afri. J. Biotechnol., 2014; 11(6): 1465-1477.
[3] Akpan I., MO Bankole and AM Adesemowo. Production of alpha amylase by Aspergillus niger in a cheap solid medium using rice bran and agricultural material. Trop. Sci., 1999; 39: 77-79.
[4] Al-Qodah Z, Daghstani H, Geopel Ph, Lafi W. Determination of kinetic parameters of α-amylase producing thermophile Bacillus sphaericus. Afr. J. Biotechnol., 2007; 6: 699-706.
[5] Asegbeloyin J.N., and Onyimonyi A.E. The effect of different processing methods on the residual cyanide of ‘Gari’. Pak. J. Nutr., 2007; 6(2): 163-166.
[6] Avwioroko, J.O. and Tonukari, N.J. Isolation and molecular identification of Aspergillus species associated with the spoilage of cassava in Nigeria. Nig. J. Sci. Environ., 2015; in press.
[7] Balkan B and Ertan F. Production and properties of α-amylase from Penicillium chrysogenum and its application in starch hydrolysis. Prep. Biochem. Biotechnol., 2005; 35: 169-178.
[8] Banea, J. P., Bradbury, J. H., Mandombi, C., Nahimana, D., Denton, I. C., and Katumbay, D. T. Effectiveness of wetting method for control of konzo and reduction of cyanide poisoning by removal of cyanogens from cassava flour. Food Nutri. Bulletin, 2014; 35(1): 28-32.
[9] Chi H.L.Z., X. Duan, L. Ma and L. Gao. Purification and characterization of extracellular amylase from the marine yeast Aureobasidium pullulans N13d and its raw potato starch digestion. Enzyme Microb. Technol., 2007; 40: 1006-1012.
[10] Chung YC, Kobayashi T, Kanai H, Akiba T, Kudo T. Purification and properties of extracellular amylase from the hyperthermophilic archaeon Thermococcus profundus DT5432. Appl. Environ. Microbiol., 1995; 61: 1502-1506.
[11] Cordeiro CAM, Martins MLL, and Luciano AB. Production and properties of α-amylase from thermophilic Bacillus sp. Braz. J. Microbiol., 2002; 33: 57-61.
[12] Dar, G.H., Kamili, A.N., Nazir, R., Bandh, S.A., and Malik, T.A. Biotechnological production of α-amylases for industrial purposes: Do fungi have potential to produce α-amylases? Int. J. Biotechnol. Mol. Biol. Res., 2014; 5(4):35-40.
[13] de Souza, P.M. and Magalhães, P.O. Application of microbial α-amylase in industry – a review. Braz. J. Microbiol., 2010; 41: 850-861.
[14] Doyle EM, Kelly CT, and Fogarty WM. The amylolytic enzymes of Penicillium amagasakiense. Biochem. Soc. Trans., 1988; 16: 181-182.
[15] Doyle EM, Kelly CT, Fogarty WM. The high maltose-producing α-amylase of Penicillium expansum. Appl. Microbiol. Biotechnol., 1989; 30:492-496.
[16] Elias, M., Wieczorek, G., Rosenne, S., and Tawfik, D. S. The universality of enzymatic rate- temperature dependency. Trends Biochem. Sci., 2014; 39(1): 1-7.
[17] Ertan F, Yagar H, and Balkan B. Some properties of free and immobilized α-amylase from Penicillium griseofulvum by solid state fermentation. Prep. Biochem. Biotechnol., 2006; 36: 81-91.
[18] Filtz, T. M., Vogel, W. K., and Leid, M. Regulation of transcription factor activity by interconnected post-translational modifications. Trends Pharmacol. Sci., 2014; 35(2): 76-85.
[19] Fogarty WM, and Kelly CT. Recent advances in microbial amylases. In: Fogarty WM, Kelly CT, (eds). Microbial enzymes and biotechnology, London and New York. 1990; Pp. 71-133.
[20] F. Nartey and B. L. Møller. Amino acid profiles of cassava seeds (Manihot esculenta). Econ. Bot., 1976; 30(4): 419-423.
[21] Gauthier, G. M., and Keller, N. P. Crossover fungal pathogens: the biology and pathogenesis of fungi capable of crossing kingdoms to infect plants and humans. Fungal Gen. Biol., 2013; 61:146-157.
[22] Gupta A., Gupta D.R Modi and Yadava L.P. Production and characterization of a-amylase from Aspergillus niger. Biotechnol., 2008; 1:1-6.
[23] Gupta, R., Gigras, P., Mohapatra, H., Goswami, V.K.,Chauhan, B. Microbial α-amylases: a biotechnological perspective. Process Biochem., 2003; 38:1599 - 1616.
[24] Hu, X., Wang, C., Wang, L., Zhang, R., and Chen, H. Influence of temperature, pH and metal ions on guaiacol oxidation of purified laccase from Leptographium qinlingensis. World J. Microbiol. Biotechnol., 2014; 30(4):1285-1290.
[25] Jadhav, S. B., Bankar, S. B., Granström, T., Ojamo, H., Singhal, R. S., and Survase, S. A. Enhanced stability of alcohol dehydrogenase by non-covalent interaction with polysaccharides. Appl. Microbiol. Biotechnol., 2014; 98(14): 6307-6316.
[26] Kathiresan K and Manivannan S. α-Amylase production by Pencillium fellutanum isolated from mangrove rhizosphere soil. Afri. J. Biotechnol., 2006; 5(10): 829-932.
[27] Khedher IBA, Bressollier P, Urdaci MC, Limam F, Marzouki MN. Production and biochemical characterization of Sclerotinia sclera otiorum α-amylase ScAmy1: assay in starch liquefaction treatments. J. Food Biochem., 2008; 32: 597-614.
[28] Kumar, D., Yadav, K. K., Muthukumar, M., and Garg, N. Production and characterization of α-amylase from mango kernel by Fusarium solani NAIMCC-F-02956 using submerged fermentation. J. Environ. Biol., 2013; 34(6): 1053-1058.
[29] Metin, K., Koc, O., Ateşlier, B. B., and Biyik, H.H. Purification and characterization of α-amylase produced by Penicillium citrinum HBF62. Afr. J. Biotechnol., 2010; 9(45):7692-7701.
[30] Miller, G.L. Use of dinitrosalicylic acid Their purification, properties, action on starch and reagent for determination of reducing sugar. Anal. Chem., 1959; 31: 426-429.
[31] Nouadri T, Meraihi Z, Shahrazed DD, and Leila B. Purification and characterization of the α-amylase isolated from Penicillium camemberti PL21. Afr. J. Biochem. Res., 2010; 4(6): 155-162.
[32] Oboh G. Isolation and characterization of amylase from fermented cassava (Manihot esculenta crantz) waste water. Afri. J. Biotechnol., 2005; 4 (10): 1117 – 1123.
[33] Ogugbue, C.J., Mbakwem-Aniebo, C. and Akubuenyi, F. Assessment of microbial air contamination of post processed garri on sale in markets. Afri. J. Food Sci., 2011; 5(8): 503 – 512.
[34] Okolo BN, Ire FS, Ezeogu LI, Anyanwu CU, Odibo FJC. Purification and some properties of a novel raw starch-digesting amylase from Aspergillus carbonarius. J. Sci. Food Agric., 2000; 81: 329-336.
[35] Osakwe, S.A. Effect of Cassava Processing Mill Effluent on Physical and Chemical Properties of Soils in Abraka and Environs, Delta State, Nigeria. Chem. Materials Res., 2012; 2 (7): 27-39.
[36] Oyewole OB and Sanni LO. Constraints in traditional cassava processing: A case of ‘fufu’ production. In: Transformation Alimentaire Du Manioc (Cassava Food Processing). Editors: Tom Agbor Egbe, AlainBrauman, Dany Griffon, Serge Treche. Institut Francais de Recherche Scientifique Pour le Development en Cooperation (ORSTOM), Paris, France. ISBN 2-7099-1279-1. 1995; Pp. 523-529.
[37] Pandey, A., Nigam, P., Soccol, C.R., Soccol, V.Y.,Singh, D. and Mohan, R. Advances in microbial amylases. Biotechnol. Appl. Biochem., 2000; 31:135–152.
[38] Roy, A., Khanra, K., Mishra, A., and Bhattacharyya, N. Partial purification and characterization of amylase from a newly isolated Bacillus megaterium Strain KAN1 from fermented rice Handia. Am. J. Curr. Microbiol., 2014; 2(1): 1-5.
[39] Selvakumar P, Ashakumary L, Helen A, and Pandey A. Purification and characterization of glucoamylase produced by Aspergillus niger in solid state fermentation. Lett. Appl. Microbiol., 1996; 23: 403-406.
[40] Sindhu R. Isolation, purification and characterization of α-amylase from Penicillium janthhinellum. Ph.D. Thesis. School of Biosciences, Mahatma Gandhi University. 2005; P. 175.
[41] Sinha, R., and Khare, S. K. Characterization of detergent compatible protease of a halophilic< i> Bacillus sp. EMB9: Differential role of metal ions in stability and activity. Biores. Technol., 2013; 145:357-361.
[42] Sun H, Zhao P, Ge X, Xia Y, Hao Z, Liu J, and Peng M. Recent advances in microbial raw starch degrading enzymes. Appl. Biochem. Biotechnol., 2010; 160: 988-1003.
[43] Tatsinkou, FB, Tavea F, Jiwoua C, Ndjouenkeu R. Screening of thermostable amylase producing bacteria and yeasts strains from some cameroonian soils. Afr. J. Microbiol. Res., 2009; 3(9): 504-514.
[44] Tatsinkou, FB, Tavea F, Jiwoua C, Ndjouenkeu R. Simultaneous production of raw starch degrading highly thermostable a-amylase and lactic acid by Lactobacillus fermentum 04BBA19. Afr. J. Biotechol., 2011; 10(34): 6564-6574.
[45] Uyoh EA, Udensio, Natwi V, and Urea I. Effect of different processing methods on cyanide content of garri from cultivars of cassava. J. Food Agric. Environ., 2007; 3(4): 105-107
[46] Vishnu, T. S., Soniyamby, A. R., Praveesh, B. V., and Hema, T. A. Production and optimization of extracellular amylase from soil receiving kitchen waste isolate Bacillus sp. VS 04. World Appl. Sci. J., 2014; 29(7): 961-967.
[47] Gornall A, Bardsmill CT, David MM. Determination of serum protein by means of biuret reaction. J. Biol. Chem., 1949; 177: 751-766.
Author Information
  • Department of Biochemistry, Faculty of Science, Delta State University, Abraka, Nigeria

  • Department of Biochemistry, Faculty of Science, Delta State University, Abraka, Nigeria

  • Department of Biochemistry, Faculty of Science, Delta State University, Abraka, Nigeria

Cite This Article
  • APA Style

    Oghenetega Jonathan Avwioroko, Nyerhovwo John Tonukari, Samuel Ogheneovo Asagba. (2015). Biochemical Characterization of Crude α-Amylase of Aspergillus spp. Associated with the Spoilage of Cassava (Manihot esculenta) Tubers and Processed Products in Nigeria. Advances in Biochemistry, 3(1), 15-23. https://doi.org/10.11648/j.ab.20150301.14

    Copy | Download

    ACS Style

    Oghenetega Jonathan Avwioroko; Nyerhovwo John Tonukari; Samuel Ogheneovo Asagba. Biochemical Characterization of Crude α-Amylase of Aspergillus spp. Associated with the Spoilage of Cassava (Manihot esculenta) Tubers and Processed Products in Nigeria. Adv. Biochem. 2015, 3(1), 15-23. doi: 10.11648/j.ab.20150301.14

    Copy | Download

    AMA Style

    Oghenetega Jonathan Avwioroko, Nyerhovwo John Tonukari, Samuel Ogheneovo Asagba. Biochemical Characterization of Crude α-Amylase of Aspergillus spp. Associated with the Spoilage of Cassava (Manihot esculenta) Tubers and Processed Products in Nigeria. Adv Biochem. 2015;3(1):15-23. doi: 10.11648/j.ab.20150301.14

    Copy | Download

  • @article{10.11648/j.ab.20150301.14,
      author = {Oghenetega Jonathan Avwioroko and Nyerhovwo John Tonukari and Samuel Ogheneovo Asagba},
      title = {Biochemical Characterization of Crude α-Amylase of Aspergillus spp. Associated with the Spoilage of Cassava (Manihot esculenta) Tubers and Processed Products in Nigeria},
      journal = {Advances in Biochemistry},
      volume = {3},
      number = {1},
      pages = {15-23},
      doi = {10.11648/j.ab.20150301.14},
      url = {https://doi.org/10.11648/j.ab.20150301.14},
      eprint = {https://download.sciencepg.com/pdf/10.11648.j.ab.20150301.14},
      abstract = {In this research, crude α-amylases associated with the spoilage of cassava (Manihot esculenta) tubers/ product (‘eba’) were biochemically characterized. They were isolated from five fungi: Aspergillus sp. CSA25, Aspergillus sp. CSA26, Aspergillus sp. CSA27, Aspergillus sp. CSA35 and Aspergillus sp. CSA38. The results of the analyses showed that the activities of α-amylase obtained from both sources (cassava tuber/ eba) were optimal at 45°C and pH 5.0. The maximum specific activity (Vmax) of the enzyme was found to be 10 U/mg protein, while its Michaelis-Menten constant (Km) was between 0.37 -1.25%w/v. The α-amylase is thermally stable for 1 - 2 h at optimum temperature and pH (45°C; pH 5.0). A broad range of substrate specificity was expressed by the enzyme for cassava starch-containing products (tapioca flour, garri flour, cassava flour, 1%, w/v); however, potato (Ipomoea batatas) starch, yam (Dioscorea rotundata) flour and cocoyam (Colocasia esculenta) flour were relatively minimally hydrolyzed by the crude α-amylases obtained from Aspergillus spp. that caused spoilage of cassava. Ethylenediamine tetraacetic acid (1 mM EDTA) and Mg2+ treatment had no significant (p > 0.05) effect on the activities of the amylase, but Na+, K+, Ca2+, Fe3+, thiourea and 5′,5′-dithiobis-2-nitrobenzoate (1 mM DTNB) enhanced its activities. The fungal α-amylases were most activated by K+ and had a salt tolerance of 1 - 2 M NaCl for 24 h. The fungal α-amylases reported in this study would find useful application in industries like food industry, detergent industry, paper industry, textile industry, pharmaceutical industry, etc where microbial α-amylases would be required for efficient and cost-effective hydrolysis of cassava starch, cassava flour and or its products.},
     year = {2015}
    }
    

    Copy | Download

  • TY  - JOUR
    T1  - Biochemical Characterization of Crude α-Amylase of Aspergillus spp. Associated with the Spoilage of Cassava (Manihot esculenta) Tubers and Processed Products in Nigeria
    AU  - Oghenetega Jonathan Avwioroko
    AU  - Nyerhovwo John Tonukari
    AU  - Samuel Ogheneovo Asagba
    Y1  - 2015/02/09
    PY  - 2015
    N1  - https://doi.org/10.11648/j.ab.20150301.14
    DO  - 10.11648/j.ab.20150301.14
    T2  - Advances in Biochemistry
    JF  - Advances in Biochemistry
    JO  - Advances in Biochemistry
    SP  - 15
    EP  - 23
    PB  - Science Publishing Group
    SN  - 2329-0862
    UR  - https://doi.org/10.11648/j.ab.20150301.14
    AB  - In this research, crude α-amylases associated with the spoilage of cassava (Manihot esculenta) tubers/ product (‘eba’) were biochemically characterized. They were isolated from five fungi: Aspergillus sp. CSA25, Aspergillus sp. CSA26, Aspergillus sp. CSA27, Aspergillus sp. CSA35 and Aspergillus sp. CSA38. The results of the analyses showed that the activities of α-amylase obtained from both sources (cassava tuber/ eba) were optimal at 45°C and pH 5.0. The maximum specific activity (Vmax) of the enzyme was found to be 10 U/mg protein, while its Michaelis-Menten constant (Km) was between 0.37 -1.25%w/v. The α-amylase is thermally stable for 1 - 2 h at optimum temperature and pH (45°C; pH 5.0). A broad range of substrate specificity was expressed by the enzyme for cassava starch-containing products (tapioca flour, garri flour, cassava flour, 1%, w/v); however, potato (Ipomoea batatas) starch, yam (Dioscorea rotundata) flour and cocoyam (Colocasia esculenta) flour were relatively minimally hydrolyzed by the crude α-amylases obtained from Aspergillus spp. that caused spoilage of cassava. Ethylenediamine tetraacetic acid (1 mM EDTA) and Mg2+ treatment had no significant (p > 0.05) effect on the activities of the amylase, but Na+, K+, Ca2+, Fe3+, thiourea and 5′,5′-dithiobis-2-nitrobenzoate (1 mM DTNB) enhanced its activities. The fungal α-amylases were most activated by K+ and had a salt tolerance of 1 - 2 M NaCl for 24 h. The fungal α-amylases reported in this study would find useful application in industries like food industry, detergent industry, paper industry, textile industry, pharmaceutical industry, etc where microbial α-amylases would be required for efficient and cost-effective hydrolysis of cassava starch, cassava flour and or its products.
    VL  - 3
    IS  - 1
    ER  - 

    Copy | Download

  • Sections