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

SBIR 第一阶段：计算机指导的高通量筛选以提高酶活性

• 批准号: 9960806
• 负责人: Joerg Bentzien
• 金额: $ 10万
• 依托单位: Xencor
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-01-01 至 2000-06-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9960806&HistoricalAwards=false
• 关键词: SBIR; Phase; Computer; Directed; Throughput

This Small Business Innovation Research Phase I project will develop an enabling technology for computer-directed high throughput screening of proteins for improved properties. By combining a fast computer screen with experimental methods, the capabilities of random library screening can be greatly extended. This technology will be tested on Bacillus circulans xylanase with the goal of improving its activity at high temperature and pH. Starting from the enzyme crystal structure, Xencor's proprietary protein design automation (PDA) technology will be used to computationally prescreen the possible sequences resulting from varying selected residue positions, thereby eliminating unfavorable sequences and thus enormously reducing the number to be screened experimentally. The result of the computer screen is an amino acid probabilitydistribution for the selected residue positions. A method will be developed to transform these probabilities into a DNA library that will be synthesized and experimentally screened for improved enzymatic activity. The computational prescreening will allow screening of ~10^80 sequences compared to 10^10 to 10^15 with conventional in vitro evolution techniques. The proposed research will result in a generally applicableprocedure that allows optimization of far more residue positions and results in the more efficient development of improved proteins by coupling rational and random methods.

这个小企业创新研究第一阶段项目将开发一种使能技术，用于计算机指导的蛋白质高通量筛选，以改善性能。通过将快速的计算机筛选与实验方法相结合，可以大大扩展随机文库筛选的能力。该技术将在环状芽孢杆菌木聚糖酶上进行测试，目的是提高其在高温和pH下的活性。从酶晶体结构开始，Xencor专有的蛋白质设计自动化（PDA）技术将用于计算预筛选可能的序列，这些序列来自不同的选定残基位置，从而消除不利的序列，从而大大减少了实验筛选的数量。计算机屏幕的结果是所选残基位置的氨基酸概率分布。将开发一种方法来将这些概率转化为DNA文库，该文库将被合成并通过实验筛选以改善酶活性。计算预筛选将允许筛选约10^80个序列，而常规体外进化技术的筛选为10^10至10^15个。拟议的研究将产生一种普遍适用的程序，允许优化更多的残基位置，并通过结合理性和随机方法更有效地开发改良蛋白质。