Assembly of chimeric glycosyltransferases for directing biosynthesis of natural products

用于指导天然产物生物合成的嵌合糖基转移酶的组装

• 批准号: BB/F023111/1
• 负责人: Peter Leadlay
• 金额: $ 44.51万
• 依托单位: University of Cambridge
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2008
• 资助国家: 英国
• 起止时间: 2008 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FF023111%2F1
• 关键词: Assembly; chimeric; glycosyltransferases; directing; biosynthesis

This project seeks to combine the efforts of two established research teams in Cambridge to solve a major outstanding problem in chemistry and biochemistry: how to use protein engineering to generate novel glycosylated compounds that may have important biological activities. A significant fraction of drugs currently used in the clinic, especially anti-infectives such as erythromycins or vancomycins, are natural products isolated from soil bacteria, or derivatives of them. The unusual sugars they contain have been shown to be vital for biological activity. Enzymic modification of natural products is an attractive option to generate new compounds and decorating compounds with different sugars is particularly appealing. One major class of glycosyltransferases (GTs), the enzymes responsible for transferring a sugar from the donor nucleotide diphosphate (NDP) sugar to the acceptor molecule, is known to be composed of two domains. One binds the NDP-sugar donor and the other the acceptor molecule. Our preliminary studies have shown that by cutting and pasting domains from different GTs, hybrid enzymes can be constructed that remain highly active and have the respective donor and acceptor specificity of the parent GTs from which they were derived. This finding has led to the synthesis of several novel vancomycin analogues. These proof of principle experiments pave the way for making a wide range of hybrid GTs from over 19,000 known parent proteins to provide a toolbox of catalysts for accessing novel compounds. Constructing and exploiting these hybrid enzymes will require diverse experimental approaches from carrying out the synthesis of the novel compounds in a test tube with the purified enzymes to genetic manipulation of the genes inside the bacterial cells targetting novel natural products. Success in this project would deliver broad potential benefits to the pharmaceutical industry.

该项目旨在联合收割机剑桥两个已建立的研究团队的努力，以解决化学和生物化学中的一个重大突出问题：如何利用蛋白质工程产生可能具有重要生物活性的新型糖基化化合物。目前在临床上使用的药物，特别是抗感染药如红霉素或万古霉素，有很大一部分是从土壤细菌中分离出来的天然产物，或它们的衍生物。它们含有的不寻常的糖已被证明对生物活性至关重要。天然产物的酶改性是产生新化合物的有吸引力的选择，并且用不同的糖修饰化合物特别有吸引力。已知糖基转移酶（GT）的一个主要类别（负责将糖从供体核苷酸二磷酸（NDP）糖转移到受体分子的酶）由两个结构域组成。一个结合NDP-糖供体，另一个结合受体分子。我们的初步研究表明，通过切割和粘贴来自不同GT的结构域，可以构建保持高度活性并具有它们所来源的亲本GT的各自供体和受体特异性的杂合酶。这一发现导致了几种新型万古霉素类似物的合成。这些原理实验的证明为从超过19，000种已知的亲本蛋白质中制备广泛的杂交GT铺平了道路，为获得新型化合物提供了催化剂工具箱。构建和利用这些杂合酶将需要多种实验方法，从在试管中用纯化的酶合成新化合物到对细菌细胞内的基因进行遗传操作以靶向新的天然产物。该项目的成功将为制药行业带来广泛的潜在利益。

项目成果

Structure of the glycosyltransferase EryCIII in complex with its activating P450 homologue EryCII.

糖基转移酶红细胞的结构及其激活的P450同源物erycii。

• DOI: 10.1016/j.jmb.2011.10.036
• 发表时间: 2012-01-06
• 期刊: JOURNAL OF MOLECULAR BIOLOGY
• 影响因子: 5.6
• 作者: Moncrieffe, Martin C.;Fernandez, Maria-Jose;Spiteller, Dieter;Matsumura, Hiroyoshi;Gay, Nicholas J.;Luisi, Ben F.;Leadlay, Peter F.
• 通讯作者: Leadlay, Peter F.