Artificial Glycosidase with Controlled Selectivity

具有受控选择性的人工糖苷酶

• 批准号: 10024717
• 负责人: Yan Zhao
• 金额: $ 29.91万
• 依托单位: IOWA STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10024717
• 关键词: 3-Dimensional; Acidity; Active Sites; Affinity; Binding; Binding Sites; Biological; Biological Assay; Biological Process; Biology; Biomimetics; Buffers; Carbohydrates; Catalysis; Chemicals; Chemistry; Complement; Complex; Development; Endoglycosidases; Enzymes; Exoglycosidases; Generations; Genomics; Glycoside Hydrolases; Glycosides; Heterogeneity; Hydrolysis; Knowledge; Lectin; Length; Methods; Modification; Molecular; Oligosaccharides; Organic Synthesis; Oxygen; Peptides; Planet Earth; Polysaccharides; Positioning Attribute; Preparation; Proteins; Proteomics; Reporting; Research Design; Shapes; Site; Structure; Techniques; Technology; Time; Water; adduct; base; biological systems; catalyst; design; functional group; human disease; imprint; improved; insight; molecular recognition; molecular size; monomer; nanoparticle; public health relevance; receptor; reconstruction; small molecule; success; sugar; surfactant; three dimensional structure; tool

Artificial Glycosidase with Controlled Selectivity Abstract Carbohydrates are the most abundant biomolecules on the earth, and involved in numerous biological processes and all major human diseases. Glycoscience, nonetheless, lags behind genomics and proteomics, due to the extreme complexity, dynamic structural diversity, and micro-heterogeneity of glycans found in biological systems. Another reason, according to the 2012 NRC report “Transforming Glycoscience”, was the lack of suitable tools and methods “to detect, describe, and fully purify glycans…and then to characterize their chemical composition and structure. Molecular recognition of carbohydrates and peptides has been long-standing challenges in bioorganic and supramolecular chemistry, due to the importance of these molecules in biology. The PI’s group has developed protein-sized molecularly imprinted nanoparticles (MINPs) to bind a wide range of biologically interesting guests including carbohydrates and peptides. They are prepared and purified in < 2 days without any special techniques, once the template, functional monomers, and cross-linkable surfactants are available. MINP-based “synthetic lectins” were shown to recognize a wide range of mono- and oligosaccharides in water with tens of micromolar binding affinities. Oligosaccharides were distinguished based on their building blocks, glycosidic linkages, and chain length. The overall objective of this proposal is to develop synthetic glycosidases with selectivities unavailable in their natural counterparts. The proposed catalysts contain substrate-specific active sites with precisely installed catalytic groups for optimal catalysis. In the traditional synthesis of receptors and supramolecular catalysts, tremendous synthetic efforts are needed just to have a binding pocket. Fine tuning of the pocket for specific and complex biomolecules is nearly impossible. The micellar imprinting technology used in the MINP preparation, on the other hand, can quickly construct multifunctionalized, complex- shaped active sites from simple building blocks. The principles to be demonstrated are not limited to glycan hydrolysis and are expected to open up many possibilities in the design and synthesis of enzyme-mimicking catalysts.

具有受控选择性的人工糖苷酶 抽象的 碳水化合物是地球上最丰富的生物分子，参与 许多生物过程和所有主要的人类疾病。尽管如此，糖科学 由于极其复杂、动态的结构，落后于基因组学和蛋白质组学 生物系统中发现的聚糖的多样性和微观异质性。另一个原因， 根据 2012 年 NRC 报告“转变糖科学”，缺乏合适的 工具和方法“检测、描述和完全纯化聚糖……然后表征 它们的化学成分和结构。 碳水化合物和肽的分子识别由来已久 由于生物有机和超分子化学的重要性，这些挑战 生物学中的分子。 PI的团队开发出了蛋白质大小的分子印迹 纳米颗粒（MINP）可结合多种生物学上感兴趣的客体，包括 碳水化合物和肽。它们在不到 2 天内制备和纯化，无需任何特殊处理 技术，一旦模板、功能单体和可交联表面活性剂 可用的。基于 MINP 的“合成凝集素”被证明可以识别多种单 以及水中具有数十微摩尔结合亲和力的寡糖。低聚糖 根据其结构单元、糖苷键和链长度进行区分。 该提案的总体目标是开发合成糖苷酶 其天然对应物不具有选择性。所提议的催化剂包含 底物特异性活性位点具有精确安装的催化基团，可实现最佳催化作用。 在传统的受体和超分子催化剂合成中，巨大的合成 仅仅为了拥有一个有约束力的口袋就需要付出努力。对口袋进行微调，以适应特定和 复杂的生物分子几乎是不可能的。胶束印迹技术应用于 另一方面，MINP 制备可以快速构建多功能、复杂的 用简单的构建块塑造活性位点。所要论证的原理并不 仅限于聚糖水解，预计将在设计中开辟许多可能性 以及模拟酶催化剂的合成。