Application of two types of CIM tube column for purification of microbial enzymes.
Chromatography conditions for two types of convection interaction media (CIM) tube monolithic column, DEAE-8 and C4-8, were investigated using three enzymes from different microorganisms. The enzymes were adsorbed on a CIM DEAE-8 tube column under the same conditions as conventional DEAE columns. The CIM C4-8 tube column required a high concentration of ammonium sulfate compared to the conventional C4 column for adsorbing the enzymes. The separation of enzymes on the CIM tube column chromatography was not affected at flow rates between 0.15 and 1.25 volumes of the column per min. Both columns were successfully applied to the purification of enzymes from crude enzyme solution. Thus, both CIM tube monolithic columns proved useful in greatly reducing the purification time, and could be used at any stage of enzyme purification.
Isobe
Kimiyasu
K
Department ofAgro-Bioscience, Iwate University, Ueda 3-18-8, Morioka 020-8550, Japan. kiso@iwate-u.ac.jp
Kawakami
Yoshimitsu
Y
D000327:Adsorption; D000426:Alcohol Dehydrogenase / Q000302:isolation & purification*; D000429:Alcohol Oxidoreductases / Q000302:isolation & purification*; D000645:Ammonium Sulfate / Q000737:chemistry; D019117:Burkholderia / Q000201:enzymology*; D002852:Chromatography, Ion Exchange / Q000295:instrumentation* / Q000379:methods; D007770:L-Lactate Dehydrogenase / Q000302:isolation & purification*
D000429:Alcohol Oxidoreductases; D000426:Alcohol Dehydrogenase; D007770:L-Lactate Dehydrogenase; C019643:alcohol oxidase; D000645:Ammonium Sulfate