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: 10710393
• 负责人: Xi Chen
• 金额: $ 31.62万
• 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/10710393
• 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; cost effective; design; glycosylation; glycosyltransferase; improved; innovation; large scale production; method development; nucleoside triphosphate; nursing mothers; 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 合成人奶寡糖（HMO）甘氨酸的化学酶结构构建 人牛奶寡糖（HMOS）构成了人类牛奶的主要组成部分，它提供了一切 那个母乳喂养的婴儿在生命的头几个月中需要。对 HMO对母乳喂养婴儿健康的多方面贡献。探索HMO的应用 婴儿配方添加剂，营养素和/或治疗药已经开始，但由于有限 以足够的量访问结构定义的HMO。已知150多个HMO的结构。 尽管努力和开发化学，酶促，化学酶和发酵方法的进步 对于HMO综合，尚未合成地实现对完整的HMO Glycome的访问。 HMO可以 从人类牛奶中纯化，但量有限。我们计划降低访问HMO的技术障碍 开发高效的用户友好糖基转移酶的化学酶方法并构建 全面的合成HMO甘氨酸。除了酶，化学酶合成策略和 开发的生产过程，将引入底物和 过程工程。将确定，设计，表征和用于合成的新酶 以系统为导向的方式的HMO。将开发用于化学酶合成途径的地图。一个 HMO的全面库，包括带有或不带有L-浓糖的分支和长链线性结构 和/或唾液酸已从人牛奶和尚未鉴定出的潜在异构体中鉴定出来的 将被构造。合成的HMO Glycome将为纯净的纯化合物标准提供 在不同的护理母亲，不同的哺乳阶段识别和分析牛奶中HMO的HMO，并在 或没有感染或其他疾病。化学酶合成过程是可以编程的，可以适应 用于自动化，并且很容易扩展，以便将来大规模生产HMO。