聚丙烯腈纤维固定化青霉素酰化酶合成头孢氨苄的研究

Studies on Enzymatic Synthesis of Cephalexin by Immobilized Penicillin Acylase by Polyacrylonitrile Fibres

将巨大芽孢杆菌胞外青霉素酰化酶通过共价键结合到聚丙烯腈纤维的衍生物上。制成的丝状固定化青霉素酰化酶表现活力达 1 5 3U g(湿重 )。固定化酶合成头孢氨苄的最适pH为 6 5 ,最适温度为 40℃。 7 ADCA的投料浓度以 4%为好 ,7 ADCA与PGME的投料量比率为1∶2 ,最佳用酶量为 1 70U g 7 ADCA。在pH6 5、温度 3 0℃时 ,固定化酶对 7 ADCA的表观米氏常数K7 ADCA为 0 1 6 2mol L ,对PGME的表观米氏常数KPGME为 0 3 6 4mol L ,最大反应速度Vmax为0 0 4 6 2mol·L- 1·min- 1,用固定化酶合成头孢氨苄 ,使用 5 0次保留酶活力 83 9%

The extracellular penicillin acylase from Bacillus megaterium was immobilized by coupling to derivatives of polyacrylonitrile fibres. The apparent activity of the immobilized enzyme was about 153U/g (wet weight). The optimal pH and temperature were 6.5 and 40℃ for synthesis of cephalexin by penicillin acylase, respectively. Whe the concentration of 7-ADCA was 4% and the ratio of PGME and 7-ADCA and 1∶2,the average velocity of synthesis reaction was highest. The optimal supplied amount of immobilized penicillin acylase was 1.125g/g 7-ADCA.. The apparent Michaelis constant for 7-ADCA was 0 162mol/L and for PGME was 0 364mol/L, Vmax was 0 0462mol·L -1·min -1 at 30℃ and pH6 5. The remained activity was about 83 9% after operating 50 times.