权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Establishing structure-led design rules for spore surface-display of enzymes of commercial interest

建立具有商业利益的酶的孢子表面展示的结构主导设计规则

基本信息

批准号：
2714262
负责人：
金额：
--
依托单位：
University of Cambridge
依托单位国家：
英国
项目类别：
Studentship
财政年份：
2022
资助国家：
英国
起止时间：
2022 至无数据
项目状态：
未结题

来源：
https://gtr.ukri.org/projects?ref=studentship-2714262
关键词：
Establishing structure led design rules

项目摘要

Bacterial (endo)spores, formed by Bacillales and Clostridiales members when nutrient starved, are the most durable cells found in nature. The composition and assembly of spores is well understood at genetic and biochemical levels, but less so at the molecular structural level. This knowledge gap is apparent in current approaches to spore surface display - molecular genetic manipulation of typically Bacillus subtilis to express and deposit heterologous proteins on the outermost spore coat. So far, only a handful of coat proteins have been successful fusion partners for heterologous proteins of interest, despite the coat containing over 70 distinct proteins. Current surface display methodology relies on trial and error, which is time consuming and often ends in failure. As such, it has failed to translate to the commercial sphere that its potential benefits merit. Recently, the structural landscape has changed through advances in successful crystallisation of spore proteins and by the generation of seemingly accurate structural models by AlphaFold. This projectaims to use this information to establish structure-led design principles that more consistently result in successful display of functional proteins on or within the spore coat. The project will be conducted with the support of Johnson Matthey and will focus on the display of enzymes of commercial interest and examining the behaviour of cells intended for spore surface display as they transition from lab to pilot scale.

细菌（内）孢子，由芽孢杆菌目和梭菌目成员在营养缺乏时形成，是自然界中发现的最持久的细胞。孢子的组成和组装在遗传和生物化学水平上得到了很好的理解，但在分子结构水平上了解较少。这种知识差距在目前的孢子表面展示方法中是明显的-通常是枯草芽孢杆菌的分子遗传操作，以在最外面的孢子外壳上表达和存款异源蛋白。到目前为止，只有少数的外壳蛋白已经成功的融合合作伙伴的异源蛋白的目的，尽管外壳包含超过70个不同的蛋白质。目前的表面显示方法依赖于试错，这是耗时的，往往以失败告终。因此，它未能将其潜在的好处转化为商业领域。最近，通过孢子蛋白质成功结晶的进展以及AlphaFold生成看似准确的结构模型，结构景观发生了变化。该项目旨在利用这些信息来建立结构导向的设计原则，从而更一致地在孢子外壳上或孢子外壳内成功展示功能蛋白。该项目将在约翰逊万丰的支持下进行，重点是展示具有商业价值的酶，并检查用于孢子表面展示的细胞从实验室规模过渡到中试规模时的行为。