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A plethora of N-glycosylation pathways in Bacteria from the epsilon Proteobacteria - a resource for glycoprospecting and toolbox for glycoengineering

来自 epsilon Proteobacteria 的细菌中的大量 N-糖基化途径 - 糖勘探的资源和糖工程的工具箱

基本信息

批准号：
BB/H017542/1
负责人：
Dennis Linton
金额：
$ 49.25万
依托单位：
University of Manchester
依托单位国家：
英国
项目类别：
Research Grant
财政年份：
2010
资助国家：
英国
起止时间：
2010 至无数据
项目状态：
已结题

来源：
https://gtr.ukri.org/projects?ref=BB%2FH017542%2F1
关键词：
plethora glycosylation pathways Bacteria epsilon

项目摘要

Glycoproteins (proteins that are modified with sugar structures) are ubiquitous biomolecules involved in most basic biological phenomena in complex living organisms such as humans, ranging from immune recognition to cancer development. They often have underestimated biological functions and in contrast to proteins and nucleic acids, glycans have escaped the cloning revolution. The experimentally tractable model bacterium Escherichia coli is often used as a 'cellular factory' to produce practically inexhaustible amounts of purified proteins for various uses. However, until recently it has not been possible to generate glycoproteins in this bacterium as these simple organisms do not make glycoproteins of this type. This how now changed. We recently identified and characterised a cluster of pgl genes which is responsible for the synthesis of glycoproteins in the simple gut bacterium Campylobacter jejuni. This is the first bacterium known to glycosylated their proteins in this way. Furthermore we have been able to transfer the segment of C. jejuni DNA containing the pgl genes into E. coli to produce recombinant glycoproteins, thus opening up the field of glycoengineering. The key enzyme in the C. jejuni pathway that couples proteins to sugars is the transferase protein termed CjPglB. Although CjPglB can transfer many sugar structures unfortunately there are many important glycostructures that it cannot. Recently we have identified dozens more bacteria that are related to C. jejuni that have different PglB sugar transferase enzymes. Indeed some of the bacteria have more than one PglB, the first time that this has been observed in bacteria, suggesting that they may have subtly different abilities to transfer different sugars and hence be invaluable for glycoengineering. In this proposal we wish to fully characterize the plethora of new PglB enzymes and their associated pathways to expand the range of genetic tools that could be used for glycoengineering. The proposal will also help answer fundamental questions as to why some bacteria require more than one PglB and the evolutionary origin of these unusual systems in bacteria. The program of work will benefit scientists interested in basic research and also in applied research particularly in the burgeoning glycobiotechnology industry.

糖蛋白（用糖结构修饰的蛋白质）是普遍存在的生物分子，涉及复杂生物体（如人类）中最基本的生物现象，从免疫识别到癌症发展。它们往往低估了生物学功能，与蛋白质和核酸相反，聚糖逃脱了克隆革命。实验上易于处理的模式细菌大肠杆菌经常被用作“细胞工厂”，以生产几乎取之不尽的各种用途的纯化蛋白质。然而，直到最近，在这种细菌中还不可能产生糖蛋白，因为这些简单的生物体不产生这种类型的糖蛋白。这怎么现在变了。我们最近确定并表征了一组pgl基因，其负责简单肠道细菌空肠弯曲杆菌中糖蛋白的合成。这是已知的第一种以这种方式使其蛋白质糖基化的细菌。此外，我们已经能够转移C的片段。将含有pgl基因的空肠DNA导入E.大肠杆菌表达重组糖蛋白，为糖工程开辟了新的领域。C.将蛋白质与糖偶联的空肠途径是称为CjPglB的转移酶蛋白。虽然CjPglB可以转移许多糖结构，但不幸的是，有许多重要的糖结构是它不能转移的。最近，我们又发现了几十种与C有关的细菌。空肠具有不同的PglB糖转移酶。事实上，一些细菌有不止一个PglB，这是第一次在细菌中观察到，这表明它们可能具有微妙的不同能力来转移不同的糖，因此对于糖工程是非常宝贵的。在该提案中，我们希望充分表征过多的新PglB酶及其相关途径，以扩大可用于糖工程的遗传工具的范围。该提案还将有助于回答一些基本问题，例如为什么一些细菌需要一个以上的PglB以及细菌中这些不寻常系统的进化起源。这项工作计划将使对基础研究和应用研究感兴趣的科学家受益，特别是在新兴的糖生物技术行业。

项目成果

期刊论文数量（6）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Functional analysis of the Helicobacter pullorum N-linked protein glycosylation system.

DOI：
10.1093/glycob/cwx110
发表时间：
2018-04-01
期刊：
Glycobiology
影响因子：
4.3
作者：
Jervis AJ;Wood AG;Cain JA;Butler JA;Frost H;Lord E;Langdon R;Cordwell SJ;Wren BW;Linton D
通讯作者：
Linton D

Additional file 2: Figure S2. of Chromosomal integration vectors allowing flexible expression of foreign genes in Campylobacter jejuni

附加文件 2：图 S2。