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.

项目摘要岩藻糖基化的人牛奶的微生物产生寡糖该提出的项目旨在为人牛奶建立高效而特定的微生物生产过程寡糖（HMO）。 HMOS是调节新生儿健康的潜在生物活性化合物发育的兴趣作为成人疾病的潜在药物治疗。 HMO是200多个类成乳为20-23 g/L的化合物，成熟牛奶中的化合物为12-14 g/l。与他们的共同前体不同乳糖，HMO被人类婴儿取消，而是通过有效改善新生儿健康抗菌和抗病毒药，益生元和炎症性免疫细胞反应级联反应的调节剂。这些 HMO的其他潜在优势使其成为预防或治疗疾病的研究目标儿童和成人。 β1-3连接的半乳糖苷GALβ3GLCNACβor称为1型聚糖，是在100多个HMO中发现的主要HMO组件。在已有的20种HMO核心结构中确定的11个包含至少一个1型聚糖终止分支。乳酸-N-tetraose（LNT，GALβ3GLCNACβ3LAC）是最简单的1类Glycan HMO。 LNT及其诱导化衍生物是最丰富的HMO之一。虽然1型聚糖结构主要在人类牛奶中，但它们的丰富程度较低（有时在其他哺乳动物的牛奶中完全不存在）。研究单个类型1的生物学功能含糖的HMO及其潜在应用，因为益生元和抗菌药物需要进入足够数量的这些结构定义的化合物。这些分子的潜力，有限自然来源的访问，以及在大规模隔离单个HMO进行研究和应用的困难激发了新型生产方法的发展。化学和体外酶合成基于当前方法的HMO对于工业规模的综合预计将是昂贵的。全细胞生物催化剂正在成为替代自我调节的生产平台，具有巨大的潜力来减少 HMO的生产成本。总体生产了短链，线性和小单核化HMO 尚未探索细胞生物催化剂，但尚未探索具有较高复杂性的结构。在这个拟议的项目中，我们将建立一种产生岩藻糖基化的HMO的策略 Fucopentaose I（LNFP I）和Lacto-n-difucosylhexaose I（lndfh I）来自乳糖和L含量在实时工程中大肠杆菌细胞。值得注意的是，我们将开发一种创新方法来控制在整个细胞系统中，糖基化为未来的其他复合HMO的微生物生产奠定了基础。