Development of Glycosynthase Enzymes to Construct Septanosyl Glycosides

构建七糖基糖苷的糖合酶的开发

• 批准号: 1506567
• 负责人: Mark Peczuh
• 金额: $ 42万
• 依托单位: University of Connecticut
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-01 至 2019-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1506567&HistoricalAwards=false
• 关键词: Development; Glycosynthase; Enzymes; Construct; Septanosyl

With this award, the Chemistry of Life Processes Program in the Chemistry Division is funding Dr. Mark Peczuh at the University of Connecticut to transform a beta-glucosyl hydrolase from Streptomyces that normally cleaves the beta-1,4-linkage between glucose residues in cellobiose into a glycosynthase that attaches a seven-membered ring septanose onto a glucose residue. Carbohydrates are biological polymers essential to numerous functions of plant and animal life. They are built up through the sequential addition of monomers by one type of enzyme or broken down by another, completely different type of enzyme; natural carbohydrates are composed primarily of monomers with a six-atom ring. This project will develop a new type of enzyme for attaching unique, seven-atom ring monomers to carbohydrates. Moreover, the new enzyme will be derived from a degradative enzyme that has been engineered to work in reverse. That is, through genetic engineering, a degradative enzyme will be converted to a synthetic enzyme. Efforts to achieve this goal will be through an international collaborative team of graduate student chemists and bioengineers. The new enzyme plus the method used to design and create it will be a training ground for a number of interdisciplinary techniques and ideas. The researchers will directly engage non-scientists to amplify the impact of the results and their enthusiasm for the research process. Glycosynthases are mutant enzymes, derived from natural glycosyl hydrolases that form rather than cleave glycosidic linkages. The project links the expertise in septanose carbohydrate synthesis and characterization in the Peczuh research group with the group of Dr. Antoni Planas at IQS in the University of Ramon Lllull in Barcelona Spain. The development process will provide information about septanose conformations, protein-septanose interactions and enzyme dynamics. Only a few glycosynthases that couple unnatural sugars to another molecule have been successfully developed which makes this investigation particularly challenging. Individual aims within the proposal represent a step-wise approach toward the overall goal of a functional glycosynthase; the development strategy is to optimize activity for the hydrolysis reaction for the new substrate and then convert the glycosidase into a glycosynthase.

有了这个奖项，化学生命过程的化学过程计划正在为康涅狄格大学的Mark Peczuh博士提供资金，以转化链霉菌的β-葡萄糖基水解酶从链霉菌中转化，该链霉菌通常将beta-1,4-链接在纤维二糖中的葡萄糖残基之间切割到糖果酶中的糖果酶中的糖浆中septa septe septe a septe a newmeme a glucte glucte a glucte a glucte a glucte glucte。 碳水化合物是对动植物生命的众多功能必不可少的生物聚合物。它们是通过通过一种类型的酶的顺序添加单体或由另一种完全不同类型的酶分解的。天然碳水化合物主要由带有六原子环的单体组成。该项目将开发一种新型的酶，用于将独特的7个原子环单体连接到碳水化合物上。此外，新酶将源自已设计为反向工作的降解酶。也就是说，通过基因工程，降解酶将转化为合成酶。实现这一目标的努力将是通过一支由研究生化学家和生物工程师组成的国际合作团队。新的酶以及用于设计和创建的方法将是许多跨学科技术和想法的训练场。研究人员将直接吸引非科学家，以扩大结果的影响及其对研究过程的热情。 糖合成酶是突变酶，源自形成而不是裂解糖苷键的天然糖基水解酶。 该项目将PECZUH研究小组中的隔膜碳水化合物合成和表征的专业知识与西班牙巴塞罗那Ramon Lllull大学的Antoni Planas博士小组联系起来。开发过程将提供有关隔糖构象，蛋白质 - 乳糖相互作用和酶动力学的信息。 只有少数几个伴随不自然糖与另一个分子的糖合成酶已经成功地开发出来，这使得这项研究特别具有挑战性。 该提案中的个体目标代表了实现功能性糖合酶的整体目标的逐步方法。开发策略是为了优化新底物的水解反应活性，然后将糖苷酶转化为糖合酶。