Microbial production of fucosylated human milk oligosaccharides

岩藻糖基化母乳低聚糖的微生物生产

基本信息

批准号：
10681333
负责人：
Shota Atsumi
金额：
$ 31.02万
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：
2022-09-01 至 2026-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10681333
关键词：
Adult Antiviral Agents Bacteria Binding Biological Process Carbohydrates Cells Chemicals Child Collaborations Colostrum Complex Derivation procedure Development Disease Engineering Entamoeba histolytica Enzymes Epithelial Cells Escherichia coli Fucose Fucosyltransferase Future Galactose Galactosides Genes Guanosine Diphosphate Fucose Helicobacter Infections Helicobacter pylori Human Human Milk Hydrophobicity Immune In Vitro Individual Industrialization Infant Inflammatory Lactose Link Mammals Methods Milk Natural Source Oligosaccharides Optics Pathway interactions Pharmacotherapy Polysaccharides Process Production Reaction Secretor blood group alpha-2-fucosyltransferase Site Structure System Therapeutic Uridine Diphosphate Galactose antimicrobial cost cost effective cytotoxicity density glycosylation improved innovation interest intestinal epithelium microbial microbial host microorganism mutant neonatal health novel novel strategies overexpression prebiotics prevent response synthetic biology tool

项目摘要

Project Summary Microbial production of fucosylated human milk oligosaccharides This proposed project aims to establish efficient and specific microbial production processes for human milk oligosaccharides (HMOs). HMOs are potent bioactive compounds that modulate neonatal health and are of interest for development as potential drug treatments for adult diseases. HMOs are a class of over 200 compounds present at 20-23 g/L in colostrum and 12-14 g/L in mature milk. Unlike their common precursor lactose, HMOs are indigestible by human infants and instead improve neonatal health by serving as effective antimicrobials and antivirals, prebiotics, and regulators of inflammatory immune cell-response cascades. These and other potential benefits of HMOs make them attractive targets of study for preventing or treating diseases in both children and adults. β1−3-Linked galactosides Galβ3GlcNAcβOR, which are called Type 1 glycans, are major HMO components found in more than 100 HMOs. Among the 20 HMO core structures that have been identified, 11 contain at least one Type 1 glycan-terminated branch. Lacto-N-tetraose (LNT, Galβ3GlcNAcβ3Lac) is the simplest Type 1 glycan HMO. LNT and its fucosylated derivatives are among the most abundant HMOs. While Type 1 glycan structures are predominant in human milk, they are less abundant (and sometimes completely absent) in the milk of other mammals. Investigating the biological functions of individual Type 1 glycan-containing HMOs and their potential applications as prebiotics and antimicrobials requires access to sufficient quantities of these structurally defined compounds. The potential of these molecules, their limited access from natural sources, and difficulty in large-scale isolation of individual HMOs for studies and applications have motivated the development of novel production methods. Chemical and in vitro enzymatic syntheses of HMOs based on current methods are expected to be costly for industrial-scale synthesis. Whole cell biocatalysts are emerging as alternative self-regulating production platforms that have significant potential to reduce the production cost of HMOs. Short-chain, linear and small monofucosylated HMOs have been produced in whole cell biocatalysts, but structures with higher complexity have not been explored. In this proposed project, we will establish a strategy for producing fucosylated HMOs including lacto-N-fucopentaose II (LNFP II), lacto-N- fucopentaose I (LNFP I) and lacto-N-difucosylhexaose I (LNDFH I) from lactose and L-fucose in live engineered Escherichia coli cells. Notably, we will develop an innovative method to control the order and the site of glycosylation in whole cell systems to lay the groundwork for future microbial production of other complex HMOs.

项目概要岩藻糖基化母乳低聚糖的微生物生产该拟议项目旨在建立高效且特定的母乳微生物生产工艺低聚糖 (HMO) 是调节新生儿健康的有效生物活性化合物。作为治疗成人疾病的潜在药物，HMO 的开发兴趣已超过 200 种。与常见的前体不同，这些化合物在初乳中的含量为 20-23 g/L，在成熟乳中的含量为 12-14 g/L。乳糖、HMO 不能被人类婴儿消化，而是通过有效地改善新生儿健康抗菌药物和抗病毒药物、益生元以及炎症免疫细胞反应级联的调节剂。 HMO 的其他潜在好处使其成为预防或治疗疾病的有吸引力的研究目标儿童和成人的 β1−3 连接半乳糖苷 Galβ3GlcNAcβOR 被称为 1 型聚糖。在 20 种 HMO 核心结构中发现了 100 多种 HMO 的主要成分。经鉴定，11 个含有至少一个 1 型聚糖末端分支（LNT、Galβ3GlcNAcβ3Lac）。是最简单的 1 型聚糖 LNT，其岩藻糖基化衍生物是最丰富的 HMO。虽然 1 型聚糖结构在母乳中占主导地位，但其数量较少（有时完全不存在）在其他哺乳动物的乳汁中研究个体1型的生物学功能。含聚糖 HMO 及其作为益生元和抗生素的潜在应用需要获得足够数量的这些结构明确的化合物的潜力，它们的有限。天然来源的获取，以及大规模分离单个 HMO 进行研究和应用的困难推动了化学和体外酶促合成的新生产方法的发展。基于当前方法的 HMO 对于工业规模的合成来说预计成本高昂。正在成为替代的自我调节生产平台，具有减少短链、线性和小单岩藻糖基化 HMO 的生产成本已经全部生产出来。细胞生物催化剂，但尚未探索更复杂的结构。在这个拟议的项目中，我们将进行探索。制定生产岩藻糖基化 HMO 的策略，包括乳-N-岩藻五糖 II (LNFP II)、乳-N- 来自活体工程改造的乳糖和 L-岩藻糖的岩藻五糖 I (LNFP I) 和乳-N-二岩藻糖基六糖 I (LNDFH I) 值得注意的是，我们将开发一种创新方法来控制大肠杆菌细胞的顺序和位点。全细胞系统中的糖基化，为未来其他复杂 HMO 的微生物生产奠定基础。