权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Enzyme Design and Catalytic Function for Production of NovelMaterials

新材料生产的酶设计和催化功能

基本信息

批准号：
8807179
负责人：
Harvey Blanch
金额：
$ 76.62万
依托单位：
University of California-Berkeley
依托单位国家：
美国
项目类别：
Continuing Grant
财政年份：
1988
资助国家：
美国
起止时间：
1988-08-01 至 1992-06-30
项目状态：
已结题

来源：
https://www.nsf.gov/awardsearch/showAward?AWD_ID=8807179&HistoricalAwards=false
关键词：
Enzyme Design Catalytic Function Production

项目摘要

This proposal describes a comprehensive program aimed at developing new methods for the production of speciality biochemicals and advanced biomaterials. These materials will be formed by reversing the normal hydrolytic action of two proteases. These proteases will be modified to alter their selectivity and to enhance their stability under reaction conditions. Trypsin or chymotrypsin specificity will be altered by several techniques; site-directed mutagenesis, screening for mutants which produce enzymes with the ability to act on the substrates of interest, refolding partially denatured enzyme around novel substrates and a new approach which will permit the insertion of synthetic amino acids at the active site of the enzyme. Cathepsin C will be modified by a refolding procedure, and immobilized by a variety of techniques. Methods for enhancing the reversal of the hydrolytic reaction for dipeptide and oligomer synthesis with non-natural amino acids will be developed. These include the use of non-aqueous solvents (both water-miscible and immiscible) to increase substrate solubility and the use of directed methods for enzyme immobilization to enhance enzyme stability under reaction conditions. Changes in enzyme conformation induced by non-aqueous solvents and by immobilization will be probed by several techniques, including EPR.

该提案描述了一个全面的方案，旨在开发新的专业生产方法生物化学品和先进的生物材料。这些材料将通过逆转两种蛋白酶的正常水解作用而形成。这些蛋白酶将被修饰以改变它们的选择性，提高它们在反应条件下的稳定性。胰蛋白酶或胰凝乳蛋白酶特异性将通过几种技术改变; 定点诱变，筛选产生具有作用于感兴趣的底物的能力的酶，在新底物周围重折叠部分变性的酶，新的方法，这将允许插入合成氨基，在酶的活性位点的酸。组织蛋白酶C将通过重折叠程序进行修饰，并通过各种技术. 用于增强水解反应的逆转的方法，用非天然氨基酸合成二肽和低聚物将发展。这些包括使用非水溶剂（两者均水混溶性和水不混溶性）以增加底物溶解度以及使用定向方法固定酶，提高酶在反应条件下的稳定性。变化非水溶剂诱导的酶构象和固定化将通过几种技术进行探测，包括 EPR。