Biochemical and Thermodynamic Characterization of β amylase from Dioscorea alata L.

M. A. Fadunsin *

Department of Biochemistry, College of Medicine, University of Lagos, Idi-Araba-Lagos, Nigeria.

O. A. T. Ebuehi

Department of Biochemistry, College of Medicine, University of Lagos, Idi-Araba-Lagos, Nigeria.

I. S. Akande

Department of Biochemistry, College of Medicine, University of Lagos, Idi-Araba-Lagos, Nigeria.

A. O. Kolawole

Department of Biochemistry, Federal University of Technology, Akure, Nigeria.

*Author to whom correspondence should be addressed.


β-amylase (E.C. is a starch hydrolyzing enzyme fondly used in foods, pharmaceuticals, and brewing industries to convert starch into maltose. The aim of this study was to determine the physicochemical, kinetic, and thermodynamic properties, as well as the potential industrial use of β-amylase from Dioscorea alata L. Studying the enzyme stability with Arrhenius methods, showed that the enzyme was stable at a temperature range of 20–500C, and had good pH stability by retaining over 50 % of its initial activity over a wide range of pH from 4 – 8 and kinetic stability by increasing the half-life of the enzyme. The activation energy (Ea) for catalysis by water yam β-amylase at 250C was 6.45kcal/mol. The activation energy (Ea), half-life, free energy change (ΔG), enthalpy change (ΔH), and entropy change (ΔS) for inactivation at optimum temperature (400C) and pH 5 were 13.92 kcal/mol, 41.25 min, 20.89 kcal/mol, 13.30 kcal/mol and -24.25 cal/mol/K respectively. Km and Vmax values were reduced from 2.25 to 2.13mg/ml and 2.95 to 1.48 µmol/min/ml respectively. The optimum pH shifted from 5 to 6, while the Optimum temperature increased from 40 to 500C after immobilization. Enzyme retained up to 67 % activity after 5 cycles. The enzyme would be of importance in manufacturing companies based on the kinetics and application features reported in this study since it is a cheap and readily available source.

Keywords: Dioscorea alata, β-Amylase, thermo-stability, half-life, immobilization, industrial application

How to Cite

Fadunsin, M. A., Ebuehi, O. A. T., Akande, I. S., & Kolawole, A. O. (2022). Biochemical and Thermodynamic Characterization of β amylase from Dioscorea alata L. Asian Journal of Research in Biochemistry, 11(1), 30–42.


Download data is not yet available.


Reddy NS, Nimmangadda A, Sambasiva RKRS. An overview of the microbial α-amylase family. African J. Biotechnol. 2003;2:645–8.

Schramm M, Loyter A. Purification of alpha-amylase by precipitation of amylase-glycogen complex. Method Enzymol. 1996; 9:533–7.

Guzmán-Maldonado H, Paredes-López O. Amylolytic enzymes and products derived from starch: A review. Crit. Rev. Food Nutr. 1995; 35:373–403.

Coutinho PM, Reilly PJ. Glucoamylase structural, functional and evolutionary relationships. Proteins. 1997;29:334–47.

Pandey A, Nigam P, Soccol CR, Socool VT, Singh D, Mohan R. Advances in microbial amylases. Biotechnol Appl. Biochem. 2000;31:1052-1058

Van der Maarel MJEC, Van der Veen B, Uitdehaag JCM, Leemhuis H, Dijkhuizen L. Properties and applications of starch converting enzymes of the alpha-amylase family. J. Biotechnol. 2002;94:137–55.

Atichokudomchai N, Jane J, Hazlewood G. Reaction pattern of novel thermostable alpha-amylase. Carbohydr Pol. 2006; 64:582–8.

Husain S, Jafri F, Saleemuddin M. Enzyme Microb. Technol. 1996; 18:275–80.

Lin KH, Fu H, Chan CH, Lo HFL, Shih MC. Generation and analyses of the transgenic potatoes expressing heterologous thermostable beta-amylase, Plant Sci. 2008; 174: 649–656.

Moura GS, Lanna EAT, Donzele JL, Falkoski DL, Rezende ST, Oliveira MGA, Albino LFT. Stability of enzyme complex solid-state fermentation Subjected to the processing of pelleted diet and storage time at different temperatures. R. Bras. Zootec. 2016;45(12):731-736.

Pervez S, Afsheen A, Samina I, Nadir NS. Shah, A.U Saccharification and liquefaction of cassava starch: an alternative source for the production of bioethanol using amylolytic enzymes by the double fermentation process. B.M.C. Biotech. 2014;14:49–59.

Gutzow I, Todorova S, Jordanov N. Kinetics of chemical reactions and phase transitions at changing temperature: general reconsiderations and a new approach, Bulg. Chem. Comm. 2010; 42:79–102.

Dos Santos JCS, Bonazza HL, De Matos LJBL, Carneiro EA, Barbosa O, Fernandez-Lafuente R, Gonçalves LRB, De Sant’Ana HB, Santiago-Aguiar RS. Immobilization of CALB on activated chitosan: application to enzymatic synthesis in supercritical and near-critical carbon dioxide, Biotechnol. Rep. 2017; 14:16–26.

Rios NS, Pinheiroa MP, Dos Santos JCS, De Fonsecacc TS, Limac LD, De Mattos MC, Freire DMG, Da Silva Júnior IJ, Rodríguez-Aguadoe E, Goncalves LRB. Strategies of covalent immobilization of a recombinant Candida antarctica lipase B on pore-expanded SBA-15 and its application in the kinetic resolution of (R, S)-Phenyl-ethyl acetate, Mol. Catal. B: Enzym. 2016;133:246–258.

Tavanoa OL, Fernandez-Lafuente R, Goulart AJ, Monti R. Optimization of the immobilization of sweet potato amylase using glutaraldehyde-agarose support. Characterization of the immobilized enzyme, Proc. Biochem. 2013;48:1054–1058.

Sheldon R.A, S. van Pelt. Enzyme immobilization in biocatalysis: why, what and how, Chem. Soc. Rev. 2013;42: 6223–6235.

DiCosimo R, McAuliffe J, Poulose AJ, Bohlmann G. Industrial use of immobilized enzymes, Chem. Soc. Rev. 2013; 42: 6437–6474.

Rodrigues RC, Hernandez K, Barbosa O, Rueda N, Garcia-Galan C, Dos Santos JCS, Berenguer-Murcia A, Fernandez-Lafuente R. Immobilization of proteins in poly-styrene-divinylbenzene matrices: functional properties and applications, Curr. Org. Chem. 2015;19:1707–1718.

Zhang G, Brokx S, Weiner JH. Extracellular accumulation of recombinant proteins fused to the carrier protein YebF in Escherichia coli. Nature Biotechnology. 2006;24(1):100-104.

Sachin Talekar and Sandeep Chavare. Optimization of immobilization of α-amylase in alginate gel and its comparative biochemical studies with free β-amylase. Recent Res. Sci. Technol. 2012;4(2):01-05.

Lonhienne, T, Gerday,C, Feller, G. Psychrophilic enzymes: revisiting the thermodynamic parameters of activation may explain local flexibility BBA. 2000; 1543:1-10.

Kolawole AO, Ajele, JO, Ravi Sirdeshmukh. Purification and Characterization of alkaline-stable β-amylase in malted African finger millet (Eleusine coracana) seed. Plant Biochemistry. 2011;46:2178-2186

Qureshi AS, Khushk I, Ali CH, Chisti Y, Ahmad A, and Majeed H. “Coproduction of protease and amylase by thermophilic Bacillus sp. BBXS-2 using open solid-state fermentation of lignocellulosic biomass,” Biocatalysis and Agricultural Biotechno-logy. 2016;8:146–151.