Carrier Free Cross-linked Peroxidase Aggregates: Synthesis and Characterization

Key Findings & Data:The initialenzyme concentration 0.05 mg/ml , BSA 5 mg and glutaraldehyde GA 4 µl/ml amounts were optimized andmaximum immobilized activities were obtained.

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Conclusion & Impact:The apparent kinetic parameters Kmapp andVmapp of cross linked enzyme aggregates have been calculated as 33.4 mM and 0.127 mmol/l.sec, respectively

🔬Design:Research Article

📍Scope:Comprehensive Sample

⚙️Variables:cross-linked enzyme aggregates · horseradish peroxidase · bovine serum albumin

Abstract

In this study, cross-linked Horseradish Peroxidase HRP aggregates were prepared. Bovin Serum Albumin BSA ,a proteic feeder, and glutaraldehyde, a bifunctional reactive, a crosslinker agent has also been used. The initialenzyme concentration 0.05 mg/ml , BSA 5 mg and glutaraldehyde GA 4 µl/ml amounts were optimized andmaximum immobilized activities were obtained. The kinetic parameters of free and immobilised enzyme have beencalculated by using Lineweaver-Burk plots. The saturation constant Km and the maximum reaction rate Vm offree enzyme were found as 2.33 mM and 0.670mmol/l.sec, respectively. The apparent kinetic parameters Kmapp andVmapp of cross linked enzyme aggregates have been calculated as 33.4 mM and 0.127 mmol/l.sec, respectively

Keywords

cross-linked enzyme aggregates horseradish peroxidase bovine serum albumin proteic feeder

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Cite This Article

TOPÇULAR, C., AYHAN, H. (2008). Carrier Free Cross-linked Peroxidase Aggregates: Synthesis and Characterization. Hacettepe Journal of Biology and Chemistry, Cilt: 36 - Sayı: 3, 255–261. https://ajindex.com/articles/256218

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References (16)

K. Mosbach, Immobilized enzymes and cells (Part B) Methods Enzymology, Vol. 135, 1987
W. Hartmeier, Immobilized Biocatalysts, Springer-Verlag, 1988
D.S. Clarc, Can immobilization be exploited to modify enzyme activity? Trends Biotechnology, 12: 439-443, 1994
J.M.S. Cabral, J.F. Kennedy, Immobilization techniques for altering thermal stability of enzymes, Thermo stability of Enzymes, edited by Gupta M. N. Berlin, Springer-Verlag, 163- 179, 1993
S. Rocchietti, A.S.V. Urritia, M. Pregnolata, A. Tagliani, J.M. Guisan, R. Fernandez-Lafuente, M. Terreni, Influence of the enzyme derivative preparation and substrate structure on the enantioselective of penicillin G acylase, Enzyme Microb. Technol, 31:88-93, 2002
L. Cao, L.V. Langen, R.A. Sheldon, Immobilised enzymes: carrier-bound or carrier –free, Current opinion in Biotechnology, 14: 387-394, 2003
N.L. St Clair, M.A. Navia, Cross-linked enzyme crystals as robust biocatalysts, J. Am. Chem. Soc., 114: 7314-7316, 1992
L. Cao, Immobilised enzymes: science or art?, Current opinion in Chemical Biology, 9: 217- 226, 2005
J. Chen, J. Zhang, B. Han, Z. Li, J. Li, X. Feng, Synthesis of cross-linked aggregates (CLEAs) in CO2-expanded micellar solutions, Colloids and Surfaces B: Biointerfaces, 48: 72-76, 2006
S.S. Caramori, K.F. Fernandes, Covalent immobilization of horseradish peroxidase onto poly(ethylene terephalate) – poly(aniline) composite, Process Biochemistry, 39: 883-888, 2004
S. Shah, A. Sharma, M.N. Gupta, Preparation of cross-linked enzyme aggregates by using bovine serum albumin as a proteic feeder, Analytical Biochemistry, 351: 207-213, 2006
R. Tyagi, R. Batra, M.N. Gupta, Amorphous enzyme aggregates: Stability towards heat and aqueous-organic co solvents mixtures, Enzyme and Microbial Technology, 24: 348- 354, 1999
B. Halphin, R. Pressey, J. Jen, N.J. Mondy, Purification and characterization of peroxidase isoenzyme from green peas ( Pisum sativum ), J.Food Sci., 54: 44-49, 1989
F. Rothstein, Differential precipitation of proteins: science and technology, In Protein Purification Process Engineering, Edited by Harrison R. G. New York: Marcel Decker, Inc, 115-208, 1994
G.H. Broun, Chemically aggregated enzymes, in; Mosbach K. (Ed.), Methods in enzymology, Academic Press, New York, 263-269, 1997
C. Topcular, H. Ayhan, Immobilisation of horseradish peroxidase onto monodisperse poly(glycidyl methacrylate) microspheres, J. Biomater. Sci. Polym