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Direct affinity immobilization of recombinant heparinase I fused to maltose binding protein on maltose-coated magnetic nanoparticles

Paper ID Volume ID Publish Year Pages File Format Full-Text
3006 147 2014 8 PDF Available
Title
Direct affinity immobilization of recombinant heparinase I fused to maltose binding protein on maltose-coated magnetic nanoparticles
Abstract

•Fe3O4@SiO2-PEO-mal was used to immobilize MBP-Hep I from the crude enzyme directly.•All Fe3O4@SiO2-PEO-mal nanoparticles exhibited excellent immobilization performance.•Immobilized MBP-Hep I showed improved thermostability (20–50 times of free MBP-Hep I).•Immobilized MBP-Hep I showed the same catalytic mechanism as free Hep I and MBP-Hep I.•Fe3O4@SiO2-PEO100k-mal-MBP-Hep I showed suitable operating stability.

Hybrid magnetic Fe3O4@SiO2-poly(ethylene oxide)-maltose (Fe3O4@SiO2-PEO-mal) nanoparticles synthesized by our group can be used as affinity adsorption carriers for direct separation of maltose binding protein-fused Hep I (MBP-Hep I) from a crude enzyme solution in a magnetic field. In this work, different PEO molecular weights for Fe3O4@SiO2-PEO-mal nanoparticles were used for characterizing of MBP-Hep I immobilization. The results showed that all four kinds of Fe3O4@SiO2-PEO-mal magnetic nanoparticles (6k, 20k, 35k and 100k for PEO) exhibited excellent adsorption capacities and the adsorption ratio increased as the PEO molecular weight increased from 6k to 100k. All four kinds of immobilized MBP-Hep I exhibited significantly improved stability at 30 °C compared with free MBP-Hep I and their half-lives were 20–50 times that of the free MBP-Hep I. Fe3O4@SiO2-PEO-mal nanoparticles with a PEO molecular weight of 100k were best able to immobilize MBP-Hep I (Fe3O4@SiO2-PEO100k-mal-MBP-Hep I). The molecular weight distribution profiles and anticoagulant activities, obtained from heparin depolymerization by free Hep I, free MBP-Hep I and Fe3O4@SiO2-PEO100k-mal-MBP-Hep I were the same. Furthermore, Fe3O4@SiO2-PEO100k-mal-MBP-Hep I exhibited reasonable reusability during enzymatic production of low molecular weight heparins (LMWHs).

Keywords
MNPs, magnetic nanoparticles; MBP, maltose binding protein; Hep I, heparinase I; LMWHs, low molecular weight heparins; CBD, cellulose binding domain; Fe3O4@SiO2-PEO-mal, Fe3O4@SiO2-poly(ethylene oxide)-maltose; Mw, weight average molecular weight; Mn, num
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Publisher
Database: Elsevier - ScienceDirect
Journal: Biochemical Engineering Journal - Volume 90, 15 September 2014, Pages 170–177
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