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Trehalose-mediated thermal stabilization of glucose oxidase from Aspergillus niger

Paper ID Volume ID Publish Year Pages File Format Full-Text
24791 43537 2009 7 PDF Available
Title
Trehalose-mediated thermal stabilization of glucose oxidase from Aspergillus niger
Abstract

Thermal inactivation and enzyme kinetics of glucose oxidase (a FAD dependent enzyme) were studied in the absence and presence of trehalose. The inactivation rate constant decreased by up to 50% at temperatures between 50 and 70 °C in the presence of 0.6 M trehalose; as a consequence the glucose oxidase half-life increased. Intrinsic fluorescence spectra showed a maximum center of spectral mass (CSM) red shift of 6.5 nm. Therefore, major structural changes seem to be related to glucose oxidase thermal inactivation. Trehalose decreased the rate constant for unfolding as monitored by CSM red shift kinetics indicating that this disaccharide favors the most compact folded state. The Ea for unfolding was increased from 204 to 221 kJ mol−1. It is proposed that FAD dissociation is preceded by the exposition of hydrophobic regions, while the presence of trehalose was able to hinder the release of FAD. Enzyme kinetics analysis showed that trehalose does not affect Vmax but instead decreases Km; as a result enzyme efficiency was increased. The stabilizing effect of trehalose in a cofactor-dependent enzyme has not been tested to date. In addition, glucose oxidase has an enormous commercial importance and therefore, the use of trehalose to stabilize glucose oxidase in its multiple applications seems to be promising.

Keywords
Glucose oxidase; Thermal inactivation; FAD dissociation; Trehalose; Fluorescence
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Trehalose-mediated thermal stabilization of glucose oxidase from Aspergillus niger
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 141, Issues 3–4, 20 May 2009, Pages 130–136
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
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us