Meshing Synthesis and Biosynthesis for KDO: A Key Plant and Bacterial Sugar

KDO（2-keto-3-deoxy-D-manno-octulosonic acid）是一种不常见的8碳酸性糖，对许多影响植物和动物的病原菌的生长至关重要。例如，在革兰氏阴性细菌如布鲁氏菌中，其导致人类和牲畜的严重感染，KDO是基于脂质A的脂多糖的关键部分。在植物和绿色藻类中也发现了KDO，尽管其在这些生物中的生物学作用仍不清楚。这种糖的高成本加上通过化学合成制备它的挑战限制了KDO的合成化学、其生物学作用以及基于抑制KDO途径的抗生素的开发的基础科学研究。该项目旨在1）优化工程努力，以从细菌来源获得克量的KDO，2）研究将KDO连接到其他结构的化学方法，3）探索在自动化溶液相寡糖合成平台中使用KDO构建模块的协议，以将这种糖纳入更大的生物活性结构，和4）测试KDO与一系列称为凝集素的植物碳水化合物结合蛋白的结合，以发现这种病原体相关糖的潜在结合配偶体。更广泛的影响：这一跨学科项目将通过吸收代表性不足群体的学生参加拟议的研究和教育活动，扩大代表性不足群体的参与;通过吸收研究生和本科生参加并鼓励他们参加国家和国际科学会议，在促进教学、培训和学习的同时，促进发现和理解;并开发出第一个基于发酵的生产工艺和一种不寻常的糖的新化学物质，这种糖对植物和动物的细菌感染性至关重要，以帮助解开感染过程并最终抑制这些感染。

KDO (2-keto-3-deoxy-D-manno-octulosonic acid) is an unusual 8-carbon acidic sugar that is essential for the growth of many pathogenic bacteria affecting plants and animals. For example, in Gram-negative bacteria such as Brucella, which causes severe infections in humans and lifestock KDO is a crucial part of the Lipid A-based lipopolysaccharides. KDO has also been discovered in plants and green algae, although its biological roles in these organisms are still unclear. The high cost of this sugar coupled with the challenge of making it by chemical synthesis has limited basic scientific studies of the synthetic chemistry of KDO, its biological roles, and the development of antibiotics based on inhibiting KDO-based pathways. This project aims to 1) optimize engineering efforts to obtain KDO in gram quantities from a bacterial source, 2) investigate chemical methods to link KDO to other structures, 3) explore protocols to use KDO building blocks in an automated solution-phase oligosaccharide synthesis platform to incorporate this sugar into larger bioactive structures, and 4) test the binding of KDO to a range of plant carbohydrate-binding proteins called lectins to discover potential binding partners of this pathogen-associated sugar. Broader Impact: This interdisciplinary project will broaden the participation of underrepresented groups by inclusion of students from underrepresented groups as participants in the proposed research and education activities; advance discovery and understanding while promoting teaching, training, and learning by inclusion of graduate and undergraduate students as participants and the encouragement of their participation at national and international scientific meetings; and develop the first fermentation-based production processes and new chemistry of an unusual sugar key to bacterial infectiousness of plants and animals to help unravel the infection processes and ultimately inhibit these infections.