| 摘要 |
PURPOSE:To sterilize a protein without causing the denaturation and decomposition of the objective protein by carrying out the heat-treatment of a fraction containing the insoluble protein, addition of a reducing agent and the subsequent removing operation prior to the activation operation of the insoluble protein produced by genetic engineering process and accumulated in cells. CONSTITUTION:An insoluble protein is produced by a gene recombination microorganism and accumulated in the microbial cell. The protein is activated by disintegrating the cell, recovering the insoluble fraction, suspending the recovered component and activating in a state capable of expressing the activity. The above process is carried out as follows. The fraction containing the protein is subjected to heat-treatment and addition of a reducing agent at an arbitrary time from the disintegration of the cell to the activation operation and the added reducing agent is removed from the above fraction after the heat- treatment and the addition of the reducing agent and before the activation operation. The reducing agent is e.g. 2-mercaptoethanol or bithioerythritol. The heat-treatment is carried out e.g. at about 70-90 deg.C for about 0.5-1hr for the sterilization of E.coli. |
