Preparation of convection interaction media isobutyl disc monolithic column and its application to purification of secondary alcohol dehydrogenase and alcohol oxidase.
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.
Isobe
Kimiyasu
K
Department of Agro-bioscience, Iwate University, Ueda 3-18-8, Morioka 020-8550, Japan. kiso@iwate-u.ac.jp
Kawakami
Yoshimitsu
Y
D000426:Alcohol Dehydrogenase / Q000737:chemistry / Q000302:isolation & purification*; D000429:Alcohol Oxidoreductases / Q000737:chemistry / Q000302:isolation & purification*; D002082:Butylamines / Q000737:chemistry; D002853:Chromatography, Liquid / Q000295:instrumentation* / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D004983:Ethanolamines / Q000737:chemistry; D015203:Reproducibility of Results; D012685:Sepharose / Q000031:analogs & derivatives / Q000737:chemistry
D002082:Butylamines; D004983:Ethanolamines; C053521:isobutylamine; C029483:Blue-Sepharose; D012685:Sepharose; D000429:Alcohol Oxidoreductases; D000426:Alcohol Dehydrogenase; C019643:alcohol oxidase; C007369:2-diethylaminoethanol