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Specific and reversible immobilization of NADH oxidase on functionalized carbon nanotubes

Paper ID Volume ID Publish Year Pages File Format Full-Text
24191 43504 2010 7 PDF Available
Title
Specific and reversible immobilization of NADH oxidase on functionalized carbon nanotubes
Abstract

Nanotechnology-inspired biocatalyst systems have attracted a lot of attention in enzyme immobilization recently. Theoretically, nanomaterials are ideal supporting materials because they can provide the upper limits on enzyme-efficiency-determining factors such as surface area/volume ratio, enzyme loading capacity and mass transfer resistance. However, common immobilization methods have limited the applicability of these biocatalysts owing to enzyme leaching, 3D structure loss, and strong diffusion resistance. Expensive enzyme purification step is also required for these methods before immobilization. In this work, we show an efficient immobilization method based on specific interaction between His-tagged NADH oxidase and functionalized single-walled carbon nanotubes without requiring enzyme purification for immobilization. We cloned the annotated NADH oxidase gene from Bacillus cereus genome and overexpressed with pET30 vector encoding N-terminal 6× His-tag. The His-tagged NADH oxidase was then immobilized onto single-walled carbon nanotubes functionalized with Nα,Nα-bis(carboxymethyl)-l-lysine hydrate. The resulting nanoscale biocatalyst has overcome the foresaid limitations, and demonstrates good loading capacity and stability while maintaining 92% maximum activity of the native enzyme. We further demonstrate that the immobilization is reversible and can retain ca. 92% activity for a couple of loading cycles.

Keywords
NADH oxidase; Single-walled carbon nanotube; Enzyme reloading; Enzyme immobilization; Nanomaterials
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Specific and reversible immobilization of NADH oxidase on functionalized carbon nanotubes
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 150, Issue 1, 1 October 2010, Pages 57–63
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