摘要
传统固定化方法常会导致酶活性大幅度下降,回收率较低,而酶定向固定由于固定化后可完全暴露其活性部位,因而可以保持较高的酶活回收率。本文主要综述了定向固定化酶的两种方法,分别为共价定向固定和非共价定向固定。其中非共价定向固定化主要是抗体与抗原、亲和素/链霉亲和素和生物素以及组氨酸标签与Co2+/Ni2+之间的亲和作用;共价定向固定化主要是通过半胱氨酸残基上的巯基与载体作用。简述了其在生物传感器、分子识别、酶生物燃料电池及酶纯化方面的应用。最后指出今后的主要研究方向为探索新的定向固定化标签以降低对酶活性部位的影响,应用新的载体以提高固定化酶酶活回收率,优化固定化过程及简化固定化步骤等。
Traditional immobilization methods often lead to substantially dropped enzyme activity,and the recovery rate is very low. However,site-specific immobilization enzymes can expose their active sites after immobilization. So it can preserve the activity,and the enzyme recovery rate is high. The site-specific immobilization is classified into two categories,one category is the non-covalent immobilization methods that take advantage of the specific interaction between avidin/streptavidin and biotin or between polyhistidine and metal ions Co2＋/Ni2＋;the other is the covalent ligation approaches--immobilization through Cys residue.The application fields include biosensors, molecular recognition,enzymatic biofuel cell and enzyme purification. The main research direction in the future is to exploration of new site-specific immobilization tags to reduce the effect on the enzyme active site,application of new carriers to improve the activity recovery of immobilized enzyme,the immobilization process optimization or simplification of immobilization steps.
出处
《化工进展》
EI
CAS
CSCD
北大核心
2013年第10期2467-2474,共8页
Chemical Industry and Engineering Progress
基金
华侨大学基本科研业务费专项基金(JB-GJ1006)
福建省高校新世纪优秀人才支持项目(07176C0)
关键词
定向固定化酶
亲和标签
共价固定
site-specific immobilized enzyme
affinity tag
covalent immobilization
