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Immobilization on magnetic nanoparticles of the recombinant trehalose synthase from Deinococcus geothermalis

Paper ID Volume ID Publish Year Pages File Format Full-Text
19133 43045 2013 6 PDF Available
Title
Immobilization on magnetic nanoparticles of the recombinant trehalose synthase from Deinococcus geothermalis
Abstract

•The obtaining magnetic nanoparticles suitable for enzyme immobilization.•Expression of trehalose synthase from Deinococcus geothermalis in E. coli host.•Immobilization of trehalose synthase with novel techniques that base on magnetic nanoparticles.•Characterization of free and immobilized new trehalose synthase from D. geothermalis.

In our study the gene encoding trehalose synthase from Deinococcus geothermalis was cloned and overexpressed in Escherichia coli Rosetta (DE3)pLysS. Wild-type trehalose synthase has been purified from host protein after cell disruption and precipitation at 20% ammonium sulphate saturation. Recombinant trehalose synthase was immobilized onto glutaraldehyde activated silanized magnetic ferrous-ferric oxide by using covalent binding method. The morphology and surface of the obtained particles were characterized using SEM. These images show that all samples have a particle size below 30 nm. The obtained immobilized preparation has specific activity of 0.134 U/g support when measured at 40 °C using maltose as substrate. Immobilization process was almost fully completed after 30 min of the reaction at 30 °C. The highest immobilization yield of the enzyme was achieved at glutaraldehyde concentration of 10 mM. No significant differences in optimal pH and temperature were observed upon immobilization. The immobilized trehalose synthase exhibited mass transfer limitation, which is reflected by higher KM and activation energy values. In addition, immobilized trehalose synthase was easily separated from the reaction medium by an external magnetic field and retained 82% of its initial activity after successive twelve repeated batches reaction.

Keywords
Trehalose synthase; Immobilization; Magnetic nanoparticles; Nanobiotechnology
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Immobilization on magnetic nanoparticles of the recombinant trehalose synthase from Deinococcus geothermalis
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Publisher
Database: Elsevier - ScienceDirect
Journal: Food and Bioproducts Processing - Volume 91, Issue 4, October 2013, Pages 632–637
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