通过超高通量筛选方法发现的辣根过氧化物酶的高度L型和D型对映体选择性变体。
Highly L and D enantioselective variants of horseradish peroxidase discovered by an ultrahigh-throughput selection method.
作者信息
Antipov Eugene, Cho Art E, Wittrup K Dane, Klibanov Alexander M
机构信息
Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
出版信息
Proc Natl Acad Sci U S A. 2008 Nov 18;105(46):17694-9. doi: 10.1073/pnas.0809851105. Epub 2008 Nov 12.
Abstract
A highly efficient selection method for enhanced enzyme enantioselectivity based on yeast surface display and fluorescence-activated cell sorting (FACS) is developed and validated. Its application to horseradish peroxidase has resulted in enzyme variants up to 2 orders of magnitude selective toward either substrate enantiomer at will. These marked improvements in enantioselectivity are demonstrated for the surface-bound and soluble enzymes and rationalized by computational docking studies.
摘要
开发并验证了一种基于酵母表面展示和荧光激活细胞分选（FACS）提高酶对映体选择性的高效筛选方法。将其应用于辣根过氧化物酶，已得到对任意一种底物对映体具有高达2个数量级选择性的酶变体。表面结合酶和可溶性酶的对映体选择性都有显著提高，并通过计算对接研究进行了合理化分析。
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