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Nano reengineering of horseradish peroxidase with dendritic macromolecules for stability enhancement

Paper ID Volume ID Publish Year Pages File Format Full-Text
17339 42661 2012 7 PDF Available
Title
Nano reengineering of horseradish peroxidase with dendritic macromolecules for stability enhancement
Abstract

A simple bio-conjugation procedure to surround a single horseradish peroxidase (HRP) enzyme molecule with dendritic polyester macromolecules (polyester-32-hydroxyl-1-carboxyl bis-MPA dendron, generation 5) was proposed. The characterization of resultant nanoparticles entitled HRP dendrozyme, was performed by transmission electron microscopy, dynamic light scattering, gel permeation chromatography and Fourier transform infrared spectroscopy. The results showed that HRP nanoparticles were spherical in shape and have an average size of 14 ± 2 nm in diameter. Furthermore, bio-conformational characterization of HRP dendrozyme was performed by means of circular dichroism and fluorescence spectroscopy to evaluate the secondary and tertiary structure changes after enzyme modification. These investigations revealed that protein conformation had small changes (in secondary and tertiary structures) after bio-conjugation. We also reported here that dendritic modification did not significantly affect the kinetic parameters of free HRP. The stabilization of HRP with dendron macromolecules as single enzyme nanoparticles resulted in improvement of half-life over 70 days storage at 4 °C as well as its tolerance under different elevated temperatures up to 80 °C and in the presence of organic solvents for 15 min. These significant results promise extensive applications of HRP particularly in harsh environmental conditions.

► Dendron macromolecules attach to HRP enzyme for stability improvement. ► Protein structures have only a few changes after bioconjugation. ► Dendron–enzyme bioconjugation causes significant thermal and organic tolerance. ► Extensive applications of this modified HRP are promised in harsh conditions.

Keywords
Horseradish peroxidase; Thermal stability; Organic stability; Dendron; Nanoparticle
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Nano reengineering of horseradish peroxidase with dendritic macromolecules for stability enhancement
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Publisher
Database: Elsevier - ScienceDirect
Journal: Enzyme and Microbial Technology - Volume 50, Issue 1, 5 January 2012, Pages 10–16
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