关键词: 定向固定化酶, 亲和标签, 共价固定

Abstract: 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.

Key words: site-specific immobilized enzyme, affinity tag, covalent immobilization