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Thermodynamic and Kinetic Studies of a-Amylase Catalyzed Reaction in Free and Carrageenan/Chitosan Immobilized State
Corresponding Author(s) : Atul Kumar
Asian Journal of Chemistry,
Vol. 26 No. 3 (2014): Vol 26 Issue 3
Abstract
The present study deals with the immobilization of a-amylase into carrageenan and chitosan medium and determination of kinetics as well as thermodynamic parameters of both free and immobilized enzyme catalyzed reaction to predict the extent of reaction and the position of equilibrium. At optimized condition, the Km value derived from Lineweaver Burk plot for free enzyme (0.37 % w/v) was lower than the immobilized enzyme (0.48 % w/v for carrageenan and 0.59 % w/v for chitosan immobilized). The free enzyme had an Ea value of 1536 cal/mol compared to those of immobilized enzyme (5463.8 cal/mol for carrageenan and 4736 cal/mol for chitosan immobilized). Computed DS value for free enzyme is more negative than the immobilized enzyme. The increasing value DGº of in immobilized enzyme system indicates that the enzyme-substrate reaction is slower during immobilization. Immobilized enzyme could be reused up to 12 days of storage.
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- G. Dey, S. Bhupinder and R. Banerjee, Braz. Arch. Biol. Technol, 46, 167 (2003); doi:10.1590/S1516-89132003000200005.
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- J.F. Kennedy, E.H.M. Melo and K. Jumel, Chem. Eng. Prog., 45, 81 (1990).
- S. Pramanik, K.K. Sarwa, A.K. Dolui and A. Kumar, Asian J. Chem., 25, 6557 (2013);doi: 10.14233/ajchem.2013.14361.
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References
A.L. Smith, Oxford Dictionary of Biochemistry and Molecular Biology, Oxford University Press, p. 43 (1997).
C.M. Grisham and R.H. Garrett, Biochemistry, Saunders College Publ., Philadelphia, edn 2, p. 426 (1999).
R.A. Messing, J. Am. Chem. Soc., 91, 2370 (1969); doi:10.1021/ja01037a030.
W.L. Stanley and A.C. Olson, U.S. Patent, 736,231 (1973).
G.R. Stark, Biochemical Aspects of Reactions on Solid Supports, Academic Press, New York (1971).
W. Gaffield, Y. Tomimatsu, A.C. Olson and E.F. Jansen, Arch. Biochem. Biophys., 157, 405 (1973); doi:10.1016/0003-9861(73)90656-5.
P. Bernfeld and J. Wan, Science, 142, 678 (1963); doi:10.1126/science.142.3593.678.
T. Yamamoto, Bacterial a-amylase (liquefying and saccharifying types) of Bacillus subtilis and related bacteria. In: The Amylase Research Society of Japan, Handbook of amylase and related enzymes. Their sources, isolation methods, properties and applications, pp. 40-45 Pergamon Press, Oxford (1988).
D.A. Fell, Portland Press, London, 1997.
R.N. Goldberg, Y.B. Tewari, D. Bell, K. Fazio and E. Anderson, J. Phys. Chem. Ref. Data, 22, 515 (1993); doi:10.1063/1.555939.
A. Tanaka and E. Hoshino, J. Biosci. Bioeng., 96, 262 (2003).
G. Dey, S. Bhupinder and R. Banerjee, Braz. Arch. Biol. Technol, 46, 167 (2003); doi:10.1590/S1516-89132003000200005.
L.M. Hamilton, C.T. Kelly and W.M. Fogarty, Carbohydr. Res., 314, 251 (1998); doi:10.1016/S0008-6215(98)00300-0.
J.F. Kennedy, E.H.M. Melo and K. Jumel, Chem. Eng. Prog., 45, 81 (1990).
S. Pramanik, K.K. Sarwa, A.K. Dolui and A. Kumar, Asian J. Chem., 25, 6557 (2013);doi: 10.14233/ajchem.2013.14361.
B. Krajewska, Enzyme Microb. Technol., 35, 126 (2004); doi:10.1016/j.enzmictec.2003.12.013.
S. Anisworth, University Park press, London, England. P. 43(1977).
M. Riaz, R. Perveen, M.R. Javed, H. Nadeem and M.H. Rashid, Enzyme Microb. Technol., 41, 558 (2007); doi:10.1016/j.enzmictec.2007.05.010.
R.S.S. Kumar, K.S. Vishwanath, S.A. Singh and A.G.A. Rao, Process Biochem., 41, 2282 (2006); doi:10.1016/j.procbio.2006.05.028.