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Immobilization of horseradish peroxidase by entrapment in natural polysaccharide
Author(s) -
Shukla Sachin P.,
Modi Kalpana,
Ghosh P. K.,
Devi Surekha
Publication year - 2003
Publication title -
journal of applied polymer science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.575
H-Index - 166
eISSN - 1097-4628
pISSN - 0021-8995
DOI - 10.1002/app.13269
Subject(s) - horseradish peroxidase , chemistry , hydroquinone , substrate (aquarium) , peroxidase , immobilized enzyme , enzyme , michaelis–menten kinetics , carrageenan , chromatography , diffusion , enzyme assay , nuclear chemistry , organic chemistry , biochemistry , oceanography , physics , thermodynamics , geology
Horseradish peroxidase (HRP), which catalyzes oxidation reduction reactions of large number of substrates, was entrapped in K‐carrageenan beads using polyethyleneimine as hardening agent. The heat and storage stability was found to be better for entrapped horseradish peroxidase than free enzyme. The entrapped enzyme showed 50% retention of its activity after 4 cycles. Effective diffusion coefficient for diffusion of hydroquinone into K‐carrageenan beads was found to be 0.27 × 10 −10 m 2 /s during enzyme‐catalyzed oxidation of hydroquinone. Kinetic parameters calculated from Lineweaver–Burke plots were observed to be K m = 8 × 10 −5 and V max = 1.53 for free enzyme, and K m = 8.3 × 10 −5 and V max = 2.18 for entrapped enzyme when enzyme concentration was kept constant and K m = 4 × 10 −11 and V max = 0.45 for free enzyme and K m = 4.5 × 10 −11 and V max = 0.58 for entrapped enzyme when substrate concentration was kept constant. This indicates that there is no conformational change during entrapment. © 2003 Wiley Periodicals, Inc. J Appl Polym Sci 91: 2063–2071, 2004