Sun W
Institute of Microbiology, Chinese Academy of Sciences, Beijing.
Chin J Biotechnol. 1994;10(2):135-44.
When bentonite I as an absorbent according to 0.6% (w/v) was added to the supernatant of the fermentation broth for adsorption of penicillin acylase from Bacillus megatherium, 100% activity of penicillin acylase and about 10% protein in the supernatant were adsorbed. The adsorption of enzyme was not obviously changed with different pH and salt concentration of the supernatant. Various kinds of buffer with different pH were used to wash the enzyme-adsorbent complex. Only 1% enzyme activity adsorbed was washed out; however, it can wash out about 15% protein adsorbed. When phosphate buffer containing 10% PEG and NaCl as an eluent was used to elute the complex, 100% of enzyme activity adsorbed on the complex would be eluted, and purification and concentration times of enzyme could achieve about 25 and 6, respectively. The isolation and purification process can be carried out at room temperature. Its characters were very simple and showed a high recovery yield of enzyme activity, and it can be directly used for isolation and purification of penicillin acylase from the fermentation broth.
当按照0.6%（w/v）将膨润土I作为吸附剂添加到发酵液的上清液中以吸附巨大芽孢杆菌的青霉素酰化酶时，上清液中100%的青霉素酰化酶活性和约10%的蛋白质被吸附。酶的吸附在上清液不同的pH值和盐浓度下没有明显变化。使用各种不同pH值的缓冲液洗涤酶 - 吸附剂复合物。仅1%被吸附的酶活性被洗出；然而，它可以洗出约15%被吸附的蛋白质。当使用含有10%聚乙二醇和氯化钠的磷酸盐缓冲液作为洗脱剂洗脱复合物时，吸附在复合物上的100%酶活性将被洗脱，并且酶的纯化和浓缩倍数分别可达到约25和6。分离和纯化过程可在室温下进行。其特点非常简单，酶活性回收率高，可直接用于从发酵液中分离纯化青霉素酰化酶。
