《生命科学》 2009, 21(3): 388-393

糖基转移酶定向进化研究进展

吴佳茜1，王　旻1*，陈代杰2，朱 丽3

（1中国药科大学生命科学与技术学院，南京 210009；2上海医药工业研究院，上海 200040；3上海来益生物药物研究开发中心有限责任公司，上海 200240）

摘　要：摘　要：糖基转移酶在改造含糖活性化合物以获得结构多样性过程中起着重要作用，定向进化不仅能够提高其对底物的宽泛性，同时也能够提高其催化活性。定向进化的成功与否取决于突变体库的多样性及筛选方法的可靠性和效率。本文简要综述了糖基转移酶随机突变方法及高通量筛选正突变体的策略，包括基于细胞的筛选法、基于荧光受体高通量筛选和pH值指示计高通量筛选。
关键词：糖基转移酶；定向进化；高通量筛选

Research advances in directed evolution of glycosyltransferase

WU Jia-qian1， WANG Min1*， CHEN Dai-jie2， ZHU Li3

(1 School of Life Science and Technology， China Pharmaceutical University， Nanjing 210009， China;2 Shanghai Institute of Pharmaceutical Industry， Shanghai 200040， China;3 Shanghai Health Creation Center for Biopharmaceutical R&D， Shanghai 200240， Chi

Abstract: Abstract: Glycosyltransferase plays an essential role in modifying bioactive compounds containing sugars. Directed evolution is one of the new strategies not only for improving catalytic activity but also expanding its promiscuity. The successful directed evolution depends on the diversity of mutant library and the high efficacy of screening method. This review briefly introduces the random mutation strategy，and the high-throughput screening（HTS） for the selection of positive mutants. The methods include a cell-based screening system， fluorescence-based acceptor HTS and pH-indicator HTS.
Key words: glycosyltransferase; directed evolution; high-throughput screening