Study on Isolation and Purification of Penicillin Acylase by Adsorption on Bentonite
新的分离纯化青霉素酰化酶方法的研究

Bentonite I as an absorbent according to 0. 65% (w/v) was added into the supernatant of fermentation broth for adsorption of penicillin acylase from Bacillus megatherium. It found that 100% activity of penicillin acylase and about 10% protein in the supernatant were adsorbed. The adsorption of enzyme was not obviously changed by different pH and salt .concentration of the supernatant. Verious kinds of buffer having different pH were used to wash the adsorbent-enzyme 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 adsorbend on the complex would be eluted, and purification and concentration of enzyme could achieve about 25 and 6 fold, respectively. The isolation and purification process can be carried out at room temperature. Its characters were very simple and high recovery yield of enzyme activity, and it can be directly used for isolation and purification of penicillin acylase from the fermentation broth.