Purification of Alcohol Dehydrogenase from Saccharomyces cerevisiae Using Magnetic Dye-Ligand Affinity Nanostructures

Reactive Green 19 was covalently immobilized onto magnetic nanostructures for purification of alcohol dehydrogenase from Saccharomyces cerevisiae . The Reactive Green 19 immobilized magnetic nanostructures were characterized by Fourier transform infrared spectroscopy, electron spin resonance, atomic...

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Bibliographic Details
Published in:Applied biochemistry and biotechnology 2013-04, Vol.169 (7), p.2153-2164
Main Authors: Kaya, Nazife, Aktaş Uygun, Deniz, Akgöl, Sinan, Denizli, Adil
Format: Article
Language:English
Subjects:
Adsorption
Alcohol
Alcohol Dehydrogenase - isolation & purification
Alcohols
Biochemistry
Biological and medical sciences
Biotechnology
Chemistry
Chemistry and Materials Science
Chromatography, Affinity - methods
Dehydrogenase
Dyes
Fourier transforms
Fundamental and applied biological sciences. Psychology
Infrared spectroscopy
Ligands
Magnetics
Nanostructured materials
Nanostructures
Nanotechnology - methods
Polymers
Saccharomyces cerevisiae
Saccharomyces cerevisiae - enzymology
Sodium chloride
Yeast
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Summary:Reactive Green 19 was covalently immobilized onto magnetic nanostructures for purification of alcohol dehydrogenase from Saccharomyces cerevisiae . The Reactive Green 19 immobilized magnetic nanostructures were characterized by Fourier transform infrared spectroscopy, electron spin resonance, atomic force microscope, and energy dispersive X-ray analysis. Particle size of nanostructures was found to be roughly 70 nm. Alcohol dehydrogenase adsorption experiments were investigated under different conditions in batch system (i.e., medium pH, alcohol dehydrogenase concentration, temperature, and ionic strength). Maximum alcohol dehydrogenase adsorption capacity was found to be 176.09 mg/g polymer while nonspecific alcohol dehydrogenase adsorption onto plain magnetic nanostructures was negligible (19.4 mg/g polymer). Alcohol dehydrogenase molecules were desorbed by using 1.0 M NaCl with 98.4 % recovery. Alcohol dehydrogenase from S. cerevisiae was purified 45.63-fold in single step with dye-immobilized magnetic nanostructures, and purity of alcohol dehydrogenase was shown by silver-stained sodium dodecyl sulfate-polyacrylamide gel electrophoresis.
ISSN:0273-2289
1559-0291
DOI:10.1007/s12010-013-0130-5