Isobe Kimiyasu, Kawakami Yoshimitsu
Department ofAgro-Bioscience, Iwate University, Ueda 3-18-8, Morioka 020-8550, Japan.
J Chromatogr A. 2005 Feb 11;1065(1):129-34. doi: 10.1016/j.chroma.2004.10.050.
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.
使用来自不同微生物的三种酶,研究了两种对流相互作用介质(CIM)管整体柱DEAE - 8和C4 - 8的色谱条件。这些酶在与传统DEAE柱相同的条件下吸附到CIM DEAE - 8管柱上。与传统C4柱相比,CIM C4 - 8管柱吸附这些酶需要高浓度的硫酸铵。在CIM管柱色谱上,酶的分离在每分钟0.15至1.25倍柱体积的流速下不受影响。两根柱子都成功应用于从粗酶溶液中纯化酶。因此,两种CIM管整体柱都被证明在大大缩短纯化时间方面很有用,并且可用于酶纯化的任何阶段。