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

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Abstract

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.

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Authors and Affiliations

  1. Faculty of Science and Arts, Chemistry Department, Adnan Menderes University, Aydin, Turkey

    Nazife Kaya & Deniz Aktaş Uygun

  2. Faculty of Science, Biochemistry Department, Ege University, Izmir, Turkey

    Sinan Akgöl

  3. Faculty of Science, Chemistry Department, Hacettepe University, Ankara, Turkey

    Adil Denizli

Authors
  1. Nazife Kaya
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  2. Deniz Aktaş Uygun
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  3. Sinan Akgöl
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  4. Adil Denizli
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Correspondence to Deniz Aktaş Uygun.

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Kaya, N., Aktaş Uygun, D., Akgöl, S. et al. Purification of Alcohol Dehydrogenase from Saccharomyces cerevisiae Using Magnetic Dye-Ligand Affinity Nanostructures. Appl Biochem Biotechnol 169, 2153–2164 (2013). https://doi.org/10.1007/s12010-013-0130-5

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  • DOI: https://doi.org/10.1007/s12010-013-0130-5

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