SBIR Phase II: Computer-Directed High Throughput Screening for Improved Enzymatic Activity

SBIR 第二阶段：计算机指导的高通量筛选以提高酶活性

• 批准号: 0091586
• 负责人: John Desjarlais
• 金额: $ 50万
• 依托单位: Xencor
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-03-15 至 2003-02-28
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0091586&HistoricalAwards=false
• 关键词: SBIR; Phase; II; Computer; Directed

This Small Business Innovation Research (SBIR) Phase II project focuses on the development of an enabling technology for computer- directed high-throughput screening of proteins with improved properties. Xencor's Protein Design Automation (PDA) predicts all the possible amino acid sequences that will fold into the three-dimensional structure of a protein. There should be molecules among those sequences that have the structure and function of the "parent "protein, together with additional novel properties such as increased thermo-stability or alkaline pH optima. In Phase I the company addressed this possibility using xylanase as a model protein. After targeting the active site of the enzyme for PDA re-design, the company found sequences that were more active than the wild-type protein and one that had a different pH profile. These results were achieved by testing only 260 of a possible 110,592 sequences. In Phase II the company will develop a high-throughput assay system that will allow testing the majority of the predicted sequences. The research will also improve electrostatic functions of the PDA algorithm, and then use this version of the program to re-design the entire xylanase molecule instead of just the active site, thereby finding mutations located away from the active site that effect the protein's characteristics.The PDA technology improves enzyme efficiency and expands the reactions and process conditions where they can be applied. Major markets include polymer manufacturers, value extraction from waste streams and food processing.

这个小企业创新研究（SBIR）第二阶段项目的重点是开发一种使能技术，用于计算机指导的高通量筛选具有改进特性的蛋白质。Xencor的蛋白质设计自动化（PDA）预测所有可能的氨基酸序列，这些序列将折叠成蛋白质的三维结构。在那些序列中应该存在具有“亲本“蛋白质的结构和功能的分子，以及额外的新特性，例如增加的热稳定性或碱性pH最适性。 在第一阶段，该公司使用木聚糖酶作为模型蛋白质来解决这种可能性。 在靶向酶的活性位点进行PDA重新设计后，该公司发现了比野生型蛋白更活跃的序列，并且具有不同的pH值。这些结果是通过测试110，592个可能序列中的260个来实现的。在第二阶段，该公司将开发一个高通量的检测系统，将允许测试大多数预测的序列。该研究还将改进PDA算法的静电功能，然后使用该版本的程序重新设计整个木聚糖酶分子，而不仅仅是活性位点，从而找到位于远离活性位点的影响蛋白质特征的突变。PDA技术提高了酶的效率，并扩展了可以应用的反应和工艺条件。 主要市场包括聚合物制造商、废物流的价值提取和食品加工。