Xian H, Wang Z
Institute of Microbiology, Chinese Academy of Sciences, Beijing 100080, China.
Wei Sheng Wu Xue Bao. 2001 Aug;41(4):475-80.
The immobilization of Penicillin G Acylase from Bacillus megaterium by glutaraldehyde crosslinking on the partially acid-hydrolyed polyacrylonitrile fiber was studied. When the amount of--NH2 on fiber were 690 mumol/g and the moisture in the fiber was 64%, and the content of enzyme protein immobilized on fiber was more than 100 mg/g. The activity of 2300 IU/g was obtained with 30% of overall yield and 56% of binding efficiency. The immobilization yield was markedly influenced by ratio of the amount of free enzyme used to the weight of the fiber. The half-life of storage stability of immobilized PGA at room temperature was 130 days. The immobilized PGA kept 80% of the initial activity after 20 cycles of operation in 10% of PGK(W/V) in 0.05 mol/L phosphate buffer, pH 8.0, at 37 degrees C and an enzyme load of 150 IU/g(PGK) and 10 g(PGK) for per cycle of operation. The hydrolysis conversion of PGK in the range of 2.5%-12.5% (W/V) were over 98% for the immobilized PGA. The operation stability of immobilized PGA treated with DTT was better than that of immobilized PGA untreated.
研究了通过戊二醛交联将巨大芽孢杆菌青霉素G酰化酶固定在部分酸水解的聚丙烯腈纤维上的方法。当纤维上的—NH₂量为690 μmol/g且纤维中的水分含量为64%时,固定在纤维上的酶蛋白含量超过100 mg/g。获得了2300 IU/g的活性,总产率为30%,结合效率为56%。固定化产率受游离酶用量与纤维重量之比的显著影响。固定化青霉素G酰化酶在室温下储存稳定性的半衰期为130天。在37℃、pH 8.0的0.05 mol/L磷酸盐缓冲液中,以150 IU/g(青霉素钾)的酶负载量和每循环10 g(青霉素钾),在10%(W/V)的青霉素钾中进行20次循环操作后,固定化青霉素G酰化酶保持了初始活性的80%。对于固定化青霉素G酰化酶,在2.5% - 12.5%(W/V)范围内的青霉素钾水解转化率超过98%。用二硫苏糖醇处理的固定化青霉素G酰化酶的操作稳定性优于未处理的固定化青霉素G酰化酶。