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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要合成人乳低聚糖 (HMO) 糖组的化学酶法构建母乳低聚糖 (HMO) 是母乳的主要成分，提供一切母乳喂养的婴儿在生命的最初几个月所需要的。人们越来越欣赏 HMO 对母乳喂养婴儿健康的多方面贡献。探索 HMO 的应用婴儿配方奶粉添加剂、营养保健品和/或疗法已经开始，但由于有限而进展缓慢获得足够数量的结构明确的 HMO。超过 150 种 HMO 的结构是已知的。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 合成，尚未通过合成实现获得完整的 HMO 糖组。 HMO 可以是从人乳中提纯，但数量有限。我们计划通过以下方式降低获得 HMO 的技术壁垒：开发高效、用户友好的基于糖基转移酶的化学酶方法并构建综合合成 HMO 糖组。除了酶之外，化学酶合成策略以及已经开发的生产工艺，将在基材和工艺工程。新酶将被识别、设计、表征并用于合成 HMO 以系统化、目标导向的方式进行。将开发化学酶合成路线图。一个综合的 HMO 库，包括含或不含 L-岩藻糖的支链和长链线性结构和/或已从母乳中鉴定出的唾液酸以及尚未鉴定出的潜在异构体将被建造。合成的 HMO 糖组将为以下物质提供充分表征的纯化合物标准品：鉴定和分析不同哺乳期母亲、不同哺乳阶段的乳汁中的 HMO 或没有感染或其他疾病。化学酶合成过程是可编程的，可以适应用于自动化，并且很容易扩展以适应未来 HMO 的大规模生产。