Carbohydrate microarray printer for plant and microbial glycomics for food, nutrition and health research

植物、酵母和细菌的细胞都被细胞壁包围，细胞壁是由碳水化合物的复杂混合物组成的。细胞壁决定了生物制品的许多重要特性，因此是遗传改良的极佳靶点。然而，分析这些碳水化合物网络非常复杂，需要数百个基因来构建它们。要正确地理解这些结构通常需要耗时和昂贵的化学分析，这使得收集足够的数据来匹配现代基因技术变得困难。为了解决这个问题，我们开发了一种“基于碳水化合物微阵列”的方法，能够分析打印在特殊显微镜载玻片上的数千个微观碳水化合物点。这种方法的微调使我们能够从数百个品种/菌株（同一物种，不同遗传）中收集细胞壁碳水化合物数据，并使我们能够将这些差异与遗传差异相匹配。将这项技术应用于IFR国家酵母培养物收藏中的酵母菌株，与诺维奇研究园区的其他团队合作，带来了令人兴奋的新研究。我们还与约克大学合作，将这项技术应用于植物。总的来说，我们目前的工作具有巨大的潜力，可以支持现有项目的一系列活动，除此之外，还可以重点关注改善人类健康、将微生物用于工业（工业生物技术）和从植物中获取可再生能源（生物能源）。不幸的是，目前我们所依赖的微阵列打印设备是旧的，无法维护。该打印机建于2004年，用于打印DNA微阵列，因此依赖于旧的软件和部件。现代碳水化合物微阵列打印机也更精确，产生的斑点质量更好。此外，我们的研究所（食品研究所）将于2018年搬到一座名为Quadram研究所的新建筑。我们已经决定，我们目前的微阵列打印机，它是一个房间的大小，不能移动，所以这个重要的工具将会丢失。本提案的目的是为新的最先进的碳水化合物微阵列打印机寻求资金，这将使我们的“糖组学”研究能够在新的QI中继续和加速。计划研究：我们的重点将反映QI和BBSRC的战略，并将集中在与食品、健康和工业应用相关的植物、酵母和其他微生物细胞壁上。利用从酵母（IFR/QI的NCYC）、油菜和小麦（约克大学）和原核微生物（东安格利亚大学和Bactevo有限公司）收集的基因序列数据，这项基于阵列的研究将致力于利用植物和微生物细胞壁来创造生化物质和“生物活性物质”，从而改善我们的健康。并了解微生物如何创造碳水化合物结构（生物膜），以保护它们免受目前医学上使用的抗微生物药物的影响。将要探索的主要研究领域将包括：(a)揭示酵母中碳水化合物变异（结构和功能）的遗传基础，继续和建立目前关于酵母和其他微生物相互作用的研究，包括在肠道中发现的研究；目的是了解可能危害人类的酵母和其他微生物（也与UEA）的行为；(b)探索植物细胞壁结构和功能中碳水化合物变化的遗传基础（与约克大学合作），以生产用于生物医学用途的新生物聚合物。例如，受控的药物输送和伤口覆盖物；(c)从酵母和其他微生物中揭示“生物膜”的生物学特性，重点是医疗安全（与工业、QI和东英吉利大学合作）。正在考虑的未来研究领域包括开发与细胞壁碳水化合物组成，结构和使用相关的其他快速和信息工具。

Plants, yeasts and bacteria cells are all surrounded by a cell wall, which is made up of a complex mixtures of carbohydrates. The cell wall determines many of the important properties of biological products and so is an excellent target for genetic improvement. However, these carbohydrate networks are very complex to analyse, and hundreds of genes are needed for their construction. To understand these structures properly generally requires time consuming and expensive chemical analyses, which can make it difficult to collect enough data to match modern genetic technologies.To address this we have developed a 'carbohydrate microarray-based' method which is able to analyse thousands of microscopic carbohydrate spots printed onto special microscope slides. Fine tuning of this approach has enabled us to collect cell wall carbohydrate data from hundreds of cultivars/strains (same species, different genetics) and has allowed us to match these differences to differences in genetics. Applying this technology to yeast strains housed at the National Collection of Yeast Cultures at IFR has led to exciting new research, working with other teams across the Norwich Research Park. We are also working with The University of York to apply this technology in plants. Overall, our current work has enormous potential to support a whole range of activities on existing projects and beyond which focus on improving Human Health, using microbes for industry (Industrial Biotechnology), and renewable energy from plants (Bioenergy). Unfortunately, the current microarray printing equipment we rely on is old and cannot be maintained. The printer was built in 2004 to print DNA microarrays and so relies on old software and parts. Modern carbohydrate microarray printers are also much more accurate and produce better quality spots. Also, our Institute (Institute of Food Research) is moving to a new building called the Quadram Institute in 2018. It has been decided that our current microarray printer, which is the size of a room, cannot be moved and so this important tool will be lost. The aim of this proposal is to seek funding for a new state-of-the-art carbohydrate microarray printer which will enable our 'Glycomics' research to continue and accelerate in the new QI. Planned Research: Our focus will reflect the strategies of the QI and the BBSRC and will concentrate on plant, yeast and other microbial cell walls relevant to food and health and industrial applications. Using genetic sequence data collected from yeasts (NCYC at IFR/QI)), oilseed rape and wheat (at University of York) and prokaryotic microbes (with University of East Anglia [UEA] and Bactevo Ltd), the array-based research will be targeted towards the use of plant and microbial cell walls for creating biochemicals and 'bioactives' which can improve our health, and understanding how microbes create carbohydrate structures (biofilms) which protect them from current anti-microbials used in medicine. The main areas of research to be explored will include: (a) Revealing the genetic basis for carbohydrate variation (structure and function) in yeasts, continuing and building on current research concerning yeast and other microbe interactions, including those found in the gut; the intention is to understand the behaviour of yeasts and other microbes (also with UEA) which can harm humans; (b) Exploring the genetic basis for carbohydrate variations in structure and functionality in cell walls of plants (with University of York) for production of new biological polymers for biomedical use. For example controlled drug delivery and wound coverings; (c) Unravelling the biology of 'biofilms' from yeasts and other microbes with a focus on medical safety (with industry, the QI and UEA). Future areas of research under consideration include development of other fast and informative tools related to cell wall carbohydrate composition, structure and use.