Statistical Optimization of Enzymatic Hydrolysis of Potato (Solanum tuberosum) Starch by Immobilized α-amylase

Enzymatic hydrolysis of starch from natural sources finds potential application in commercial production of alcoholic beverage and bioethanol. In this study the effect of starch concentration, temperature, time and enzyme concentration were studied and optimized for hydrolysis of Potato starch powder (of mesh 80/120) into glucose syrup by immobilized (using Sodium arginate) α-amylase using central composite design. The experimental result on enzymatic hydrolysis of Potato starch was subjected to multiple linear regression analysis using MINITAB 14 software. Positive linear effect of starch concentration, enzyme concentration and time was observed on hydrolysis of Potato starch by α-amylase. The statistical significance of the model was validated by F-test for analysis of variance (p ≤ 0.01). The optimum value of starch concentration, enzyme concentration, temperature, time and were found to be 6% (w/v), 2% (w/v), 40°C and 80min respectively. The maximum glucose yield at optimum condition was 2.34 mg/mL.

Authors:

• N.Peatciyammal
• B.Balachandar
• M.Dinesh Kumar
• K.Tamilarasan
• C.Muthukumaran

Keywords:

• Alcoholic beverage
• Central Composite Design
• Enzymatic hydrolysis
• Glucose yield
• Potato Starch.

References:

[1] F.W. Schenck, R.E. Hebeda, "Starch Hydrolysis Products: Worldwide
Technology, Production and Applications", vol.45, New York: VCH
publishers, 1993.
[2] Gangadharan, D., Nampoothiri, K. M., Sivaramakrishnan, S., & Pandey,
A. Biochemical characterization of raw-starch-digesting alpha amylase
purified from Bacillus amyloliquefaciens. Applied Biochemistry and
Biotechnology, vol.3, pp.653-62, 2009.
[3] Woolfe, J.A, "The potato in the human diet", UK: Cambridge University
Press, 1987.
[4] Hasirci, N., Aksoy, S., & Tumturk, H, "Activation of poly(dimer acidco-
alkyl polyamine) particles for covalent immobilization of a-amylase",
Reactive and Functional Polymers, vol.66, pp.1546-1551,2006.
[5] Markweghanke, M., Lang, S., & Wagner, F, "Dodecanoic acid inhibition
of a lipase from Acinetobacter sp. OPA 55", Enzyme and Microbial
Technology, vol.17, pp.512-516, 1995.
[6] Mozhaev, V. V., Khmelnitsky, Y. L., Sergeeva, M. V., Belova, A. B.,
Klyachko, N. L., Levashov, A. V., et al. "Catalytic activity and
denaturation of enzymes in watedorganic cosolvent mixtures", European
Journal of Biochemistry, vol.184, pp.597,1989.
[7] Rajagopalan, G., & Krishnan, C, "Immobilization of maltooligosaccharide
forming a-amylase from Bacillus subtilis KCC103:
Properties and application in starch hydrolysis", Journal of Chemical
Technology and Biotechnology, vol.83 pp.1511-1517, 2008.
[8] Reshmi, R., Sanjay, G., & Sugunan, S, "Enhanced activity and stability
of amylase immobilized on alumina", Catalysis Communications, vol.7,
pp.460-465, 2006.
[9] T.E. Fannin, M.D. Marcos, D.A. Anderson, H.L. Bergman, "Use of
fractional factorial design to evaluate interactions of environmental
factors affecting biodegradation rates", Appl. Environ. Microbiol., vol.42
pp.936-943, 1981.
[10] Adinarayana.K, P. Ellaiah, B. Srinivasulu, R. Bhavani Devi, G.
Adinarayana,"Response surface methodological approach to optimize
the nutritional parameters for neomycin production by Streptomyces
marinensis under solid-state fermentation", Process Biochem., vol.38,
pp.1565-1572, 2003.
[11] M. Elibol, D. Ozer, "Response surface analysis of lipase production by
freely suspended Rhizopus arrhizus", Process Biochem.,vol.38, pp.367-
372, 2002.
[12] Tamilarasan.K, A.Subramanian, M.Dharmendira Kumar, "Optimization
of Enzymatic Hydrolysis of Rice Starch by Immobilized ╬▒-Amylase
using Response Surface Methodology", IJBST., vol.3, pp. 61-67, 2010.
[13] Johnsen A, Flink JM, "Influence of alginate properties and gel
reinforcement on fermentation characteristics of immobilized yeast
cells", Enz Microb Technol., vol.8, pp.737-748, 1986.
[14] Miller, G. L, "Use of dinitro-salicylic acid reagent for determination of
reducing sugars", Anal. Chem., vol.31, pp.426-428, 1959