Williams Gavin J, Zhang Changsheng, Thorson Jon S
Laboratory for Biosynthetic Chemistry, Pharmaceutical Sciences Division, School of Pharmacy, National Cooperative Drug Discovery Program, University of Wisconsin-Madison, 777 Highland Avenue, Madison, Wisconsin 53705, USA.
Nat Chem Biol. 2007 Oct;3(10):657-62. doi: 10.1038/nchembio.2007.28. Epub 2007 Sep 9.
Natural products, many of which are decorated with essential sugar residues, continue to serve as a key platform for drug development. Adding or changing sugars attached to such natural products can improve the parent compound's pharmacological properties, specificity at multiple levels, and/or even the molecular mechanism of action. Though some natural-product glycosyltransferases (GTs) are sufficiently promiscuous for use in altering these glycosylation patterns, the stringent specificity of others remains a limiting factor in natural-product diversification and highlights a need for general GT engineering and evolution platforms. Herein we report the use of a simple high-throughput screen based on a fluorescent surrogate acceptor substrate to expand the promiscuity of a natural-product GT via directed evolution. Cumulatively, this study presents variant GTs for the glycorandomization of a range of therapeutically important acceptors, including aminocoumarins, flavonoids and macrolides, and a potential template for engineering other natural-product GTs.
天然产物，其中许多都带有必需的糖残基，仍然是药物开发的关键平台。在这类天然产物上添加或改变连接的糖可以改善母体化合物的药理特性、多个层面的特异性，甚至作用的分子机制。尽管一些天然产物糖基转移酶（GTs）具有足够的通用性，可用于改变这些糖基化模式，但其他酶的严格特异性仍然是天然产物多样化的限制因素，并凸显了对通用GT工程和进化平台的需求。在此，我们报告了基于荧光替代受体底物的简单高通量筛选方法的使用，通过定向进化来扩大天然产物GT的通用性。累积来看，本研究展示了用于一系列具有治疗重要性的受体（包括氨基香豆素、黄酮类化合物和大环内酯类）糖基随机化的变体GT，以及工程改造其他天然产物GT的潜在模板。
