Investigation into Optimal Conditions for Enzymatic Hydrolysis of Cassava Starch to Glucose by Amylase from Rice
DOI:
https://doi.org/10.54536/ajsts.v2i2.1763Keywords:
Amylase, Cassava, Glucose, Hydrolysis, RiceAbstract
The challenge of finding locally available materials in abundance to meet up with the increase in demand for glucose syrup necessitated this study. Enzymatic hydrolysis of cassava starch to glucose using glucose amylase sourced from rice was conducted. A-3 factor with 6 levels factorial viz. substrate concentration (0.5, 1.0, 1.5, 2.0, 2.5 and 3.0% w/v), pH (4, 5, 6, 7, 8 and 9) and temperature (30, 40, 50, 60, 70 and 80 °C) experiment were employed. Rice malt was prepared and enzyme activated. Starch (substrate), buffer solutions, standard glucose solution and its calibration curve were also prepared. Starch was hydrolyzed by α-amylase and tested for presence of reducing sugar using Benedict solution. Time course of the reaction was studied and enzyme activity determined. It was observed that as reaction time increased (t), amount of glucose produced [P] initially increased but soon recorded infinitesimal increase and later assumed constant. The effects of substrate concentration, pH and temperature were found to be essential on glucose production. Statistical analysis on the effect of substrate concentration [S], reaction time and their interactions showed significant impact at probability level of (p) = 0.05.
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Adejumo, B. A. and Ola, F. A. (2010). The Effect of Cassava Effluent on the Chemical Composition of Agricultural Soil Engineering. 2nd Edition. Academic Press Inc., New York, 250p.
Adenise, L.W., Saul, N. I., Ashock, P. and Carlos, S. (2002). Acid and enzymatic hydrolysis to recover reducing sugars from cassava. Baggasse: An economic study. Brazil Archive of Biotechnology ,45(3), 1516-1522.
Alias-Rodianah, B. (2009). The Effect of Substrate and Enzyme Concentration to the Fructo- oligosaccharides (FOS) Production. BEng Project. Faculty of Chemical & Natural Resources Engineering, University Malaysia Pahang, Malaysia, 125p. (Thesis).
AnalChem-Resources (2023). Preparation of pHBuffersolutions http://delloyd.50megs.com/MOBILE/buffersolns.html (Online).
AOAC (Association of Official and Analytical Chemists) (1998). Official Methods of Analysis. 13th Edition, Association of Official and Analytical Chemists, Washington D.C. 876p.
Bailey, J. E. and Ollis, D. F. (1986). Biochemical Engineering Fundamentals. 2nd Edition. Mc Graw Hill, United Kingdom, 928p.
Balagopalan, C. (2002). Cassava utilization in food, feed and industry. In: R. J. Hillocks, J. M. Tresh and A.C. Belloti (Editors) Cassava: Biology, Production and Utilization. CIAT, Columbia, 301-318.
Cochran, B., Lunday, D. and Miskevich, F. (2008). Kinetic Analysis of Amylase Using Quantitative Benedict’s and Iodine Starch Reagents. Journal of Chemical Education, 85(3), 401- 404.
Cornish- Bowden, A. (2014). Fundamental of Enzymes Kinetics Revised Edition, Elsevier Science, London, 173p.
Edu-Enzyme (2018). Enzyme Activity Determination. http://sites.psu.edu/kmf5429/ lab3 enzyme (Retrieved on 10th May, 2018).
Eric, B. (2017). Recent progress. In: Ian, J. T.(Editor) Understanding Starch Structure. Agronomy, 2(7), 56, https://doi.org/10.3390/agronomy7030056.
Franco, C. M. and Ciacco, C.F. (1992). Factors that affect the enzymatic degradation of natural starch granules: Effect of the size of the granules. Starch Starke, 44 (11), 422-426.
Franco, C. M., Ciacco, C. F. and Tavares, D. Q. (1988). Studies on the susceptibility of granular cassava and corn starches to enzymatic attack. Part 2: Study of the granular structure of starch. Starch-Starke, 40(1), 29-32.
Geetha, D. K. (2012). Practical Biochemistry. Jaypee Brothers Medical Publishers Ltd, New Delhi: 4 - 7.
Hammond,T. K. and Ayernor, G. S. (2000). Combination of malted series and cassava starch in the production of sugar syrup. Journal of the Ghana Science Association, 2(1), 87-92.
Hoover, R. and Zhou,Y. (2003). Invitro and invivo hydrolysis of legume starches by alpha amylase and resistance starch for formation in legumes: A review. Carbohydrate Polymers, 54(4), 401-417.
Jain, S. K. and Verma, R. C. (2003). Osmotic dehydration: A new, promising and emerging industry. Beverage Food World, 30, 30–34.
Jane, J. (2006). Current understanding on starch granule structure. Journal of Applied Glycoscience, 53(3), 205-213.
Jasco International (2019). Application Note: Measurement of Enzyme Activity - Alkaline Phosphatase Activity. http://www.jasco corporation.com (Retrieved on August, 2019).
Karolina, A. B. (2015). Kinetic Modelling of Enzymatic Starch Hydrolysis. PhD Thesis, Wageningen University, Wageningen, 160p. (Thesis).
Khan, S. (2013). Saying Meritocracy and Doing Privilege: The Sociological Quarterly. http://www.wileyonlinelibrary.com (Retrieved on 25th August, 2019).
Kolusheva, T. and Marinova, A. (2007). A study of the optimal conditions for starch hydrolysis through thermostable α-amylase. Journal of the University of Chemical Technology and Metallurgy, 42(1), 93-96.
Li, J. H. (2004). Starch from hull-less barley: In-vitro susceptibility of waxy normal and high amylase starches towards hydrolysis by alpha amylases and amyloglucosidase. Food Chemistry, 84(4), 621-634.
Megazyme (2019). D-glucose Assay Procedure. www.megazyme.com (Retrieved on 19th April 2020).
Nam, S. W. (2023). Starch Hydrolysis by Amylase: Experiment NO. 5 https://user.eng.umd.edu/~nsw/ench485/lab5.htm (Retrieved on 7th June 2023). (Online).
Nieto, A. B., Salvatori, D. M., Castro, M. A. and Alzamora, S. M. (2004). Structural changes in apple tissue during glucose and sucrose osmotic dehydration: shrinkage, porosity, density and microscopic features. Journal of Food Engineering, 61(2), 269 – 278.
Nor, S. (2009). Effects of pH and Temperature on Glucose Production from Tapioca Starch Using Enzymatic Hydrolysis: A Statistical Approach. BEng. Faculty of Chemical and Natural Resources Engineering University Malaysia, Pahang, Malaysia, 24p. (Thesis).
Odebunmi, E. O. and Owalude, S. O. (2005). Kinetics and mechanism of oxidation of sugars by chromium (VI) in perchloric acid medium. Journal Chemical Society of Nigeria, 30(2), 187-191.
Omojola, M.O., Manu, N. I., Thomas, S. A. (2011). Effect of acid hydrolysis on the physicochemical properties of colar starch. African Journal of Pure and Applied Chemistry, 5 (9), 307-315.
Onyenekwe, P.C. (2013). Glucose Syrup Production. RMRDC Proceedings Series 025: Adoption of Improved Processing Equipment for Cassava Cluster Development in CRCRI Adopted Villages, Abuja, Nigeria, 123p. (Proceedings).
Osipina, B. and Wheatley, C. (2005). Processing of Cassava Tubers, Meals and Chips. http://www.fao.org (Retrieved on 24th July 2019). (Proceedings).
Oyewole, O.B. and Obieze, N. (1995). Processing and characteristics of tapioca meal from cassava. Tropical Science, 35, 401-404.
Ozen, B. F., Dock, L. L. Ozdemir, M. and Floros, J. D. (2002). Processing factors affecting the osmotic dehydration of diced green peppers. International Journal of Food Science and Technology, 37, 497–502.
Ponto, J. and Low, N.H. (1995). Glucose syrup production from Indonesian palm and cassava starch. Food Research International, 28(4), 379-385.
Rebecca, V. A., Wout, B. and Ruben, V. (2016). Saccharification protocol for small-scale lignocellulosic biomass samples to test processing of cellulose into glucose. Bioprotocol, 6(1), 1 -10 https://www.researchgate.net/publication/308753308 (Retrieved 13th March, 2018). (Online).
Rosa, M. D. and Giroux, F. (2001) Osmotic treatments and problems related to the solution management. Journal of Food Engineering, 49, 223 – 236.
Sciencell (2019). Glucose Assay. Science Cell Research Laboratories. (Book). www.sciencellonline.com (Retrieved 11th April 2020).
Silva, R. N., Fabio, P. Q., Valdirene, N. M., Eduardo, R. A., Ciencia, T. A. (2008). Production of glucose and fructose syrups from cassava (Manihot esculanta Crantz) starch using enzymes produced by microorganisms isolated from Brazilian Cerrado soil. The Journal of Biological Chemistry, 30.
Tello, P.G., Rubio, F.C., Alameda, E. J. and Rodriguez, R.S. (1993). Kinetics of the hydrolysis of soluble starch by glucoamylase. International Chemical Engineering, 33 (3), 450- 455.
Tester, R.F., Qi, X. and Karkalas, J. (2006). Hydrolysis of native starches with amylase. Animal Feed Science and Technology, 130, 39-54.
Torreggiani, D. and Bertolo, G. (2001). Osmotic pretreatments in food processing: Chemical, physical and structurals effects. Journal of Food Engineering, 49, 247-253.
Tunde, A. A. (2020). Production of glucose from hydrolysis of potato starch. World Scientific News, 145, 128-143.
Uk Essays (2018). Starch Hydrolysis by Amylase: An Experiment https://www.ukessays.com/essays/biology/the-starch-hydrolysis-of-amylase (Retrieved 11th April 2020).
WBC (Worthington Biochemical Corporation) (2020). Introduction to Enzymes. http://www.worthington-biochem.com/introbiochem/enzymeconc.html (Retrieved on 29th April 2022).
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