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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

Functionalization of paramagnetic nanoparticles for protein immobilization and purification

Full text
Author(s):
Carneiro, Lara A. B. C. [1] ; Ward, Richard J. [2]
Total Authors: 2
Affiliation:
[1] Univ Sao Paulo, Fac Med Ribeirao Preto, Dept Bioquim & Imunol, BR-14049900 Ribeirao Preto, SP - Brazil
[2] Univ Sao Paulo, Fac Filosofia Ciencias & Letras Ribeirao Preto, Dept Quim, Ave Bandeirantes 3900, BR-14049901 Ribeirao Preto, SP - Brazil
Total Affiliations: 2
Document type: Journal article
Source: Analytical Biochemistry; v. 540, p. 45-51, JAN 1 2018.
Web of Science Citations: 5
Abstract

A paramagnetic nanocomposite coated with chitosan and N-(5-Amino-1-carboxy-pentyl) iminodiacetic acid (NTA) that is suitable for protein immobilization applications has been prepared and characterized. The nanoparticle core was synthesized by controlled aggregation of Fe3O4 under alkaline conditions, and Transmission Electron Microscopy revealed a size distribution of 10-50 nm. The nanoparticle core was coated with chitosan and derivatized with glutaraldehyde and NTA, as confirmed by Fourier Transform Infrared Spectroscopy. The final nanoparticles were used as a metal affinity matrix to separate a recombinant polyhistidine-tagged beta-galactosidase from Bacillus subtilis directly from E. colt cell lysates with high purity (> 95%). After loading with Ni2+, nanoparticles demonstrated a binding capacity of 250 jig of a polyhistidine-tagged beta-galactosidase per milligram of support. The immobilized enzyme retained 80% activity after 9 cycles of washing, and the immobilized recombinant protein could be eluted with high purity with imidazole. The applications for these nanomagnetic composites extend beyond protein purification, and can also be used for immobilizing enzymes, where the beta-galactosidase immobilized on the nanomagnetic support was used in multiple cycles of catalytic reactions with no significant loss of catalytic activity. (AU)

FAPESP's process: 10/18850-2 - Identification, characterization and engineering of plant cell wall degrading enzymes
Grantee:Richard John Ward
Support Opportunities: Research Projects - Thematic Grants
FAPESP's process: 16/15708-7 - Characterization of novel hydrolysis products from the synergistic hydrolysis of xyloglucan by a beta-galactosidase/alpha-xylosidase mixture
Grantee:Lara Aparecida Buffoni de Campos Carneiro
Support Opportunities: Scholarships abroad - Research Internship - Doctorate
FAPESP's process: 12/24147-8 - Rational design of multifunctional enzymes for depolimerization of lignocelulosic material
Grantee:Lara Aparecida Buffoni de Campos Carneiro
Support Opportunities: Scholarships in Brazil - Doctorate