Preparation of convection interaction media isobutyl disc monolithic column and its application to purification of secondary alcohol dehydrogenase and alcohol oxidase

J Chromatogr A. 2007 Mar 9;1144(1):85-9. doi: 10.1016/j.chroma.2006.10.088. Epub 2006 Nov 20.

Abstract

A convection interaction media (trade name CIM, BIA Separation, Ljubljana, Slovenia) isobutyl monolithic disc was prepared by incubating a CIM epoxy monolithic disc with isobutylamine, and it was then applied to the purification of secondary alcohol dehydrogenase (S-ADH) and primary alcohol oxidase (P-AOD). Both enzymes were adsorbed on this column and eluted with high purity. Thus, S-ADH was purified to an electrophoretically homogeneous state by four column chromatographies using CIM DEAE-8 and CIM C4-8 tube monolithic columns, blue-Sepharose column and CIM isobutyl disc monolithic column. P-AOD was also purified to an electrophoretically homogeneous state by three column chromatographies of CIM DEAE-8 tube, CIM C4-8 tube and CIM isobutyl disc columns.

MeSH terms

  • Alcohol Dehydrogenase / chemistry
  • Alcohol Dehydrogenase / isolation & purification*
  • Alcohol Oxidoreductases / chemistry
  • Alcohol Oxidoreductases / isolation & purification*
  • Butylamines / chemistry
  • Chromatography, Liquid / instrumentation*
  • Chromatography, Liquid / methods*
  • Electrophoresis, Polyacrylamide Gel
  • Ethanolamines / chemistry
  • Reproducibility of Results
  • Sepharose / analogs & derivatives
  • Sepharose / chemistry

Substances

  • Butylamines
  • Ethanolamines
  • isobutylamine
  • Blue-Sepharose
  • Sepharose
  • Alcohol Oxidoreductases
  • Alcohol Dehydrogenase
  • alcohol oxidase
  • 2-diethylaminoethanol