Affordable sialoglycans and associated reagents for expanded chemoenzymatic production

用于扩大化学酶生产的经济实惠的唾液酸聚糖和相关试剂

• 批准号: 10693333
• 负责人: Lim Andrew Lee
• 金额: $ 65.34万
• 依托单位: INTEGRATED MICRO-CHROMATOGRAPHY SYSTEMS, INC.
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-15 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10693333
• 关键词: Acids; Amines; Animals; Bacterial Infections; Basic Science; Binding Proteins; Biological; Biotinylation; Carbon; Cell surface; Chemicals; Collaborations; Communities; Complex; Cytolysis; Diagnostic; Disaccharides; Disease; Enzymes; Equilibrium; Event; Family; Family member; Fermentation; Future; Glycolipids; Glycoproteins; Goals; Homeostasis; Homologous Gene; Human; Human Biology; Human Milk; Human Pathology; Human body; Hydrophobicity; In Situ; Industry; Infrastructure; Malignant Neoplasms; Mammals; Methods; Molecular Probes; Monitor; Monosaccharides; N-Acetylneuraminic Acid; Neoplasm Metastasis; Oligosaccharides; Pathogenesis; Pathologic Processes; Physiological Processes; Play; Polysaccharides; Price; Process; Production; Productivity; Property; Protein Engineering; Protocols documentation; Reaction; Reaction Time; Reagent; Research; Role; Sales; Sialic Acids; System; Techniques; Testing; Therapeutic; Vertebral column; Virus Diseases; candidate identification; commercialization; commercialization readiness; cost; glycosylation; glycosyltransferase; human disease; immunoregulation; improved; innovation; large scale production; mannosamine; manufacture; mutant; research and development; sugar; sugar nucleotide; therapeutic development

Project Summary Sialoglycans are sialic acid-containing oligosaccharides that play important biological roles in human biology and pathology and are indispensable molecular probes for research related to bacterial and viral infection, cancer metastasis, immune regulation, etc. The multifaceted functions of sialoglycans in human milk are also being explored. Nevertheless, sialoglycan-related research and development of therapeutics and diagnostics have been hampered by the limited access to these structurally complex compounds as well as the high cost and special expertise needed for synthesizing and purifying these reagents. Among various synthetic strategies for producing sialoglycans, glycosyltransferase-based chemoenzymatic methods are attractive strategies. An efficient method for synthesizing structurally defined sialoglycans is one- pot multienzyme (OPME) chemoenzymatic strategy developed by Dr. Hai Yu and Dr. Xi Chen at UC Davis. Each OPME glycosylation reaction contains a glycosyltransferase and enzymes involved in generating the corresponding sugar nucleotide donor in situ from a simple monosaccharide. The OPME reactions allow the synthesis of target glycans containing the desired glycosidic linkage with high regio- and stereo-specificity. At the current stage, large-scale synthesis by the OPME systems is still limited by the scales and the yields of the enzymes produced. In addition, OPME reaction conditions can be improved to decrease the amounts of enzymes used for the synthesis. Product purification processes can also be streamlined using innovative strategies. To advance sialic acid-related research and therapeutic development, the goal of this proposed project is to develop reagents, enzymes, and methods that are ready for commercialization to allow low-cost access to sialoglycans of high demands and their underlying asialoglycans by the broad scientific community (both academic and industry) and to enable highly efficient production of diverse biologically important special sialoglycan targets by even nonspecialists. This will be achieved by the collaboration of the UC Davis team with IMCS which has expertise in high-yield enzyme production by fermentation, unique lysis technique, commercial infrastructure to manufacture, and product sale at affordable prices to the community.

项目摘要 唾液酸聚糖是含唾液酸的寡糖，在人类生物学中发挥重要的生物学作用， 是研究细菌和病毒感染、癌症、癌症和癌症相关疾病不可或缺的分子探针。 转移，免疫调节等。母乳中唾液酸聚糖的多方面功能也正在研究中。 探讨了尽管如此，唾液酸聚糖相关的研究和治疗和诊断的开发已经取得了进展。 由于获得这些结构复杂的化合物的途径有限以及成本高， 合成和纯化这些试剂所需的特殊专业知识。 在用于产生唾液酸聚糖的各种合成策略中，基于糖基转移酶的化学酶促 方法是有吸引力的策略。合成结构确定的唾液酸聚糖的有效方法是一种- 罐多酶（OPME）化学酶策略由加州大学戴维斯分校的Hai Yu博士和Xi Chen博士开发。每个 OPME糖基化反应含有糖基转移酶和参与产生OPME糖基化的酶。 从简单单糖原位合成相应的糖核苷酸供体。OPME反应允许 合成含有具有高区域和立体特异性的所需糖苷键的靶聚糖。在 现阶段，OPME系统的大规模合成仍然受到规模和产量的限制。 生产的酶。此外，可以改进OPME反应条件以减少酶的量 用于合成。产品纯化过程也可以使用创新策略进行简化。 为了推进唾液酸相关的研究和治疗开发，本项目的目标是 开发试剂，酶和方法，准备商业化，使低成本获得 广泛的科学界对高需求的唾液酸聚糖及其基础的去唾液酸聚糖（两者都 学术界和工业界），并能够高效生产各种生物学上重要的特殊 唾液酸聚糖的目标，即使是非专业人士。这将通过加州大学戴维斯分校团队与 IMCS拥有通过发酵生产高产量酶的专业知识，独特的裂解技术，商业 基础设施的生产，和产品销售以负担得起的价格向社会。