Chemoenzymatic construction of synthetic human milk oligosaccharide (HMO) glycome

合成人乳低聚糖 (HMO) 糖组的化学酶法构建

• 批准号: 10567752
• 负责人: Xi Chen
• 金额: $ 31.6万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-22 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10567752
• 关键词: Atlases; Automation; Binding Proteins; Breastfed infant; Carbohydrates; Chemicals; Discipline of Nursing; Disease; Economics; Engineering; Enzymes; Escherichia coli; Fermentation; Fucose; Future; Health; Human Milk; In Situ; Individual; Infant formula; Infection; Isomerism; Lactation; Lactose; Libraries; Life; Maps; Methods; Milk; Monosaccharides; Nutraceutical; Oligosaccharides; Polysaccharides; Process; Production; Reagent; Role; Route; Sialic Acids; Site; Source; Speed; Structure; Substrate Specificity; Therapeutic; Vertebral column; base; cost effective; design; glycosylation; glycosyltransferase; improved; innovation; large scale production; method development; nucleoside triphosphate; nursing mothers; programs; scale up; sialylation; sugar nucleotide; tool; user-friendly

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.

项目摘要 人工合成人乳寡糖(HMO)糖基的化学酶法构建 人乳低聚糖(HMOS)是人乳的主要成分，提供一切 母乳喂养的婴儿在他们生命的头几个月所需要的。人们越来越多地认识到 保健组织对母乳喂养婴儿健康的多方面贡献。HMOS在AS中的应用探索 婴儿配方奶粉添加剂、营养食品和/或疗法已经开始，但由于有限的原因，进展缓慢 获得足够数量的结构性界定的医疗保健组织。目前已知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并取得了进展 对于HMO的合成，还没有合成到一个完整的HMO糖基。HMO可以是 从母乳中提纯，但数量有限。我们计划通过以下方式降低获得医疗保健服务的技术门槛 建立基于糖基转移酶的高效、易用的化学酶方法，并构建 全面的合成HMO甘露糖。除了酶，化学酶的合成策略，以及 在已经开发的生产工艺中，将针对基材和 过程工程学。新的酶将被识别、设计、表征并用于合成 以系统的、以目标为导向的方式开展保健组织的工作。将开发化学酶合成路线的地图。一个 含或不含L-岩藻糖的包括支链和长链线形结构的HMO综合文库 和/或已从母乳中鉴定出的唾液酸和尚未鉴定的潜在异构体 将会被建造。合成的HMO糖精将为 鉴定和分析不同哺乳母亲、不同哺乳期和不同乳汁中的HMOS 或者没有感染或其他疾病。化学酶法合成过程可编程，可调 用于自动化，并且可随时扩展以用于未来HMO的大规模生产。