仿生硅化固定化葡萄糖氧化酶与辣根过氧化酶研究

Immobilization of glucose oxidase and horseradish peroxidase in biomimetic silica particles

采用分步法将仿生硅化与脂质体技术相结合,模拟细胞微结构,以卵磷脂为原料,制备脂质体微囊,以胺类为诱导剂,在其表面硅化形成氧化硅壳层,实现对葡萄糖氧化酶(GOx)与辣根过氧化酶(HRP)双酶固定。研究了超声时间、诱导剂PDADMA用量、硅前驱体水解液(TMOS)用量、Triton X-100浓度等因素对固定化酶活性影响。结果表明,在200μL PDADMA、0.7 m L TMOS、超声50 min制得的固定化酶活性最高,能迅速硅化形成直径200 nm左右的球形微囊。在此优化条件下制备的固定化酶经重复使用7次,依然达到初始催化酶活的61.46%,经一个月冷藏后酶活仍能达到最初的74.1%。可见,经过固定化,双酶系统的稳定性得到了显著提高。

The fractional step method was applied to immobilize GOx and HRP by the combination of biomimetic silicification and liposome technology, using amine as an inducer, liposome as template.The time of ultrasonic, the dosage of PDADMA, the dosage of TMOS and the concentration of TritonX-100 on the effect of enzyme activity were studied.The results showed that the immobilized enzyme prepared with 200 p,L PDADMA,0.7 mL TMOS, 1% Triton X-100 and 50 min for ultrasound had the highest activity.The results of SEM showed that immobilized enzyme particles were near-spheres,which were about 200 nm.The immobilized enzyme prepared under the optimal conditions could be of higher stability and recovery rate. The immobilized enzyme repeated seven cycles could still retain 61.46% of its initial activity.In the storage of the 30 days, the relative activity of immobilized GOx could still retain 74.1% of its initial activity. Thus, the stability of the double enzyme system has been significantly improved after immobilization.