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Pressure-induced covalent immobilization of enzymes onto solid surface

Paper ID Volume ID Publish Year Pages File Format Full-Text
4066 206 2009 5 PDF Available
Title
Pressure-induced covalent immobilization of enzymes onto solid surface
Abstract

Here in, we demonstrate a rapid method for covalent immobilization of enzymes onto activated polystyrene microtiter plate by application of pressure. Thus, when HRP, taken in an activated well of a polystyrene microtiter plate, is subjected to an optimum pressure of 2.0 × 105 Pa for an optimum time of 25 min in a closed chamber, it shows more than 5-fold increase in absorbance value compared to control experiment carried out without applying pressure. Pressure-induced immobilized HRP shows better thermal and storage stability with respect to free enzyme. Reusability study shows that immobilized enzyme retained almost full activity after three successive uses; however activity falls to half after ten cycles of repeated use. The Michaelis–Menten constant (Km) and maximum reaction velocity (Vmax) for pressure-induced immobilized HRP are found to be 55.3 μM and 0.957 mM/min, respectively, whereas for free enzyme the corresponding values are 34.7 μM and 1.089 mM/min. The method is found to be equally effective for immobilization of other enzymes including glucose oxidase, alkaline phosphatase and invertase. As no thermal incubation is required, pressure-induced immobilization could be used for immobilization of heat sensitive biomolecules useful for preparation of biosensors, biochips and protein-microarrays.

Keywords
Covalent immobilization; Activated surface; Enzyme immobilization; Horseradish peroxidase; Polystyrene; Pressure
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Publisher
Database: Elsevier - ScienceDirect
Journal: Biochemical Engineering Journal - Volume 48, Issue 1, 15 December 2009, Pages 136–140
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
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Full-text PDF Download
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