Chemoenzymatic construction of synthetic human milk oligosaccharide (HMO) glycome

项目摘要 化学酶法构建合成人乳低聚糖（HMO）糖组 人乳低聚糖（HMO）是人乳的主要组成部分， 母乳喂养的婴儿在生命的最初几个月需要的营养。人们越来越认识到， 健康维护组织对母乳喂养婴儿健康的多方面贡献。探索健康维护组织的应用， 婴儿配方食品添加剂、营养品和/或治疗剂已经开始，但由于有限的 获得足够数量的结构定义的HMO。已知150多种HMO的结构。 尽管在开发化学、酶、化学酶和发酵方法方面做出了努力和取得了进展， 对于HMO的合成，还没有合成地获得完整的HMO糖组。HMO可以是 从人乳中纯化，但量有限。我们计划通过以下措施， 开发高效的用户友好的基于糖基转移酶的化学酶促方法，并构建一个 综合合成HMO糖体。除了酶，化学酶合成策略， 已经开发的生产工艺，将为基材引入额外的创新， 工艺工程新的酶将被鉴定、工程化、表征并用于合成 健康维护组织以系统的、有针对性的方式。将绘制化学酶合成路线图。一 HMO的综合文库，包括含或不含L-岩藻糖的支链和长链线性结构 和/或唾液酸，以及尚未鉴定的潜在异构体 将被建造。合成的HMO糖组将提供良好表征的纯化合物标准品， 鉴定和分析不同哺乳期母亲、不同泌乳阶段的乳汁中的HMO， 或没有感染或其他疾病。化学酶促合成过程是可编程的，可以适应 用于自动化，并且易于扩展用于未来HMO的大规模生产。

Project Summary Chemoenzymatic construction of synthetic human milk oligosaccharide (HMO) glycome Human milk oligosaccharides (HMOs) constitute a major component of human milk which provides everything that breast-fed infants need in the first several months of their lives. There is an increasing appreciation of the multifaceted contribution of HMOs to the health of breast-fed infants. Exploring the applications of HMOs as infant formula additives, nutraceuticals, and/or therapeutics has begun but has been slow due to the limited access to structurally defined HMOs in sufficient amounts. The structures of more than 150 HMOs are known. Despite efforts and advances in developing chemical, enzymatic, chemoenzymatic, and fermentation methods for HMO synthesis, the access to a complete HMO glycome has not been achieved synthetically. HMOs can be purified from human milk but the amount is limited. We plan to lower the technical barrier to access HMOs by developing highly efficient user-friendly glycosyltransferase-based chemoenzymatic methods and construct a comprehensive synthetic HMO glycome. Other than the enzymes, the chemoenzymatic synthetic strategies, and the production processes that have been developed, additional innovation will be introduced for substrate and process engineering. New enzymes will be identified, engineered, characterized, and used for the synthesis of HMOs in a systematic target-oriented manner. Maps for chemoenzymatic synthetic routes will be developed. A comprehensive library of HMOs including branched and long-chain linear structures with or without L-fucose and/or sialic acid that have been identified from human milk and potential isomers that have not been identified will be constructed. The synthetic HMO glycome will provide well characterized pure compound standards for identifying and profiling of HMOs in the milk from different nursing mothers, at different lactation stages, and with or without infections or other diseases. The chemoenzymatic synthetic process is programmable, can be adapted for automation, and is readily scalable for large-scale production of HMOs in the future.