基于荧光激活细胞分选技术的超高通量酶活性筛选方法及其应用

基于荧光激活细胞分选(FACS)技术的超高通量酶活性筛选方法是新出现的一类高通量筛选技术.它利用流式细胞仪高灵敏度、高通量的特点,能以极高的速度(108/天)对大容量酶基因文库进行筛选.FACS筛选技术的出现突破了常规筛选方法低效、耗时、费力等瓶颈问题,极大地提升了人类对大容量基因文库的探索能力,因此在新酶基因筛选、酶活性检测、酶定向进化等领域有广泛的应用潜力.综述了FACS超高通量酶活性筛选方法的最新研究进展,着重介绍了其在酶定向进化中的应用.

Ultrahigh-throughput Enzymatic Screening Method Based on Fluorescence-activated Cell Sorting and Its Applications

Due to its super sensitivity and incomparable high throughput, fluorescence-activated cell sorting (FACS) based screening method is emerging as an important ultrahigh-throughput enzymatic screening technology recently. It could detect multiple fluorescent parameters simultaneously and screen enzyme libraries at an extremely high speed ( > 108/d), thus possesses many advantages over the conventional technologies. FACS based screening method greatly improves our ability in handling large protein libraries and has tremendous potential in various fields such as metagenomics and directed evolution. Herein, recent progress of FACS based enzymatic screening method has been reviewed, with an emphasis on its applications in enzyme directed evolution.