Exploring the biotechnological potential of GH68 family fructosyltransferases from evolutionary diverse haloarchaea

探索来自进化多样的盐古菌的 GH68 家族果糖基转移酶的生物技术潜力

• 批准号: 2460047
• 金额: --
• 依托单位: University of St Andrews
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2020
• 资助国家: 英国
• 起止时间: 2020 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2460047
• 关键词: Exploring; biotechnological; potential; GH68; family

Fructans such as inulins and levans are fructose-based oligo- and polysaccharides derived from sucrose that have found use in a variety of industrial, agricultural and healthcare applications, most notably as prebiotic dietary supplements. Previously, it was thought that fructan biosynthesis was limited to bacteria and plants. However, recent results have for the first time demonstrated fructan biosynthesis by archaeal organisms, specifically halophilic archaea (haloarchaea). This PhD project will focus on characterising the enzymes and mechanisms of fructan biosynthesis by haloarchaea with a view to establishing new modalities for fructan biosynthesis that can be deployed by industrial biotech. The haloarchaea are a particularly attractive platform for biotech applications. They are non-hazardous, genetically tractable (allowing facile heterologous gene expression), straightforward to grow in high salt medium at mesophilic temperatures and produce enzymes that are active in high salt and generally stable at elevated temperatures. The project will focus initially on haloarchaeal GH68 family fructosyltransferase (FT) enzymes, with four primary objectives: [a] to engineer heterologous expression of evolutionarily diverse haloarchaeal GH68 enzymes in the well-characterised genetically tractable (but non-fructogenic) haloarchaeon Haloferax volcanii, [b] to develop molecular genetic tools allowing regulated high level expression of haloarchaeal GH68 enzymes in their natural host organisms, [c] to purify and characterise in vitro the activities of selected evolutionarily diverse haloarchaeal GH68 enzymes, up to and including protein structure determination by X-ray crystallography, and [d] to utilise the newly developed molecular genetic tools and protocols to analyse the cellular function of GH68 enzymes via reverse genetic approaches. The project brings together the expertise of the MacNeill lab in culturing and genetically manipulating haloarchaea with the expertise of the Gloster lab in the enzymology and structural biology of carbohydrate processing enzymes.

果蛋白和levans之类的果糖是基于果糖的寡糖和源自蔗糖的多糖，它们在各种工业，农业和医疗保健应用中都有使用，最著名的是作为益生饮食补充剂。以前，人们认为果糖生物合成仅限于细菌和植物。但是，最近的结果首次证明了古细菌，特别是卤素古细菌（Haloarchaea）。该博士学位项目将着重于表征Haloarchaea果糖生物合成的酶和机制，以期建立可以通过工业生物技术部署的果糖生物合成的新方式。 Haloarchaea是生物技术应用的特别有吸引力的平台。它们是非危害的，可遗传的，可容纳异源基因表达），在中等温度下直接在高盐培养基中生长，并产生活性在高盐中并且在升高温度下稳定的酶。 The project will focus initially on haloarchaeal GH68 family fructosyltransferase (FT) enzymes, with four primary objectives: [a] to engineer heterologous expression of evolutionarily diverse haloarchaeal GH68 enzymes in the well-characterised genetically tractable (but non-fructogenic) haloarchaeon Haloferax volcanii, [b] to develop molecular genetic tools allowing受调节的卤代GH68酶在其天然宿主生物中的高度高表达，[c]以纯化和表征体外表征所选进化上多种多样的卤代GH68酶的活性，直至和包括蛋白质结构确定蛋白质结构，包括X射线结晶的蛋白质结构，并通过[d]利用新的元素属，以利用GH6的固有遗传功能，以分析分析，分析，分析，以分析为单位，方法。该项目汇集了麦克尼尔实验室在培养和遗传操纵卤素方面的专业知识，并在碳水化合物加工酶的酶学和结构生物学中使用Gloster Lab的专业知识。