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.

项目概要 唾液酸聚糖是含唾液酸的寡糖，在人类生物学和 病理学，是细菌和病毒感染、癌症相关研究不可缺少的分子探针 转移、免疫调节等。母乳中唾液酸聚糖的多方面功能也正在被研究 探索过。尽管如此，唾液酸聚糖相关的治疗和诊断研究和开发已经取得进展。 由于获取这些结构复杂的化合物的机会有限以及成本高和 合成和纯化这些试剂所需的特殊专业知识。 在生产唾液酸聚糖的各种合成策略中，基于糖基转移酶的化学酶法 方法是有吸引力的策略。合成结构确定的唾液酸聚糖的一种有效方法是： 罐式多酶（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 糖基化反应包含糖基转移酶和参与生成 从简单的单糖原位产生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目标聚糖。在 现阶段，OPME系统的大规模合成仍受到规模和产量的限制。 产生的酶。此外，可以改善OPME反应条件以减少酶的用量 用于合成。产品纯化过程也可以使用创新策略来简化。 为了推进唾液酸相关的研究和治疗开发，该项目的目标是 开发可商业化的试剂、酶和方法，以低成本获得 广泛的科学界对高要求的唾液酸聚糖及其潜在的唾液酸聚糖（两者 学术界和工业界）并能够高效生产多种具有重要生物学意义的特殊物质 即使是非专业人士也可以确定唾液酸聚糖的目标。这将通过加州大学戴维斯分校团队与 IMCS 在发酵高产酶生产、独特的裂解技术、商业化方面拥有专业知识 制造基础设施，以及以可承受的价格向社区销售产品。