Improving Glycosyltransferase by RACHITT Gene Shuffling

通过 RACITT 基因改组改进糖基转移酶

• 批准号: 6404741
• 负责人: YONGQUAN XUE
• 金额: $ 10万
• 依托单位: ACERA BIOSCIENCES, INC.
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-09-01 至 2002-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6404741
• 关键词: Streptomyces; bacterial proteins; biological products; biotechnology; catalyst; enzyme activity; genetic manipulation; glycosyltransferase; lactones; macrolide antibiotics; protein engineering; recombinant proteins

DESCRIPTION (provided by applicant): Glycosyltransferases involved in linking polyketide macrolactones with deoxysugars are key enzymes for combinatorial biosynthesis of novel macrolide anti-infective agents. In this proposal, a new gene shuffling technology, RACHITT, is applied to the desosarninyl transferase (Des VII) from the pikromycin biosynthetic pathway of Streptomyces venezuelae. Des VII is an enzyme with remarkable inherent substrate flexibility, and we expect to expand further its substrate spectrum to include 16-membered ring macrolactones. This technology is expected to provide glycosyltransferase enzymes that can catalyze the biosynthesis of 12-, 14-, and 16-membered ring macrolides. Moreover, we expect to generate a recombinant DesVII that has a higher enzymatic activity toward the 14-membered ring macrolactone narbonolide. This will be a key step in combinatorial biosynthesis of ketolide antibiotics that have a potential to combat multi-drug resistant bacterial pathogens. PROPOSED COMMERCIAL APPLICATION: NOT AVAILABLE

描述(申请人提供)：参与连接的糖基转移酶 含有脱氧糖的聚酮大内酯是组合酶的关键酶 新型大环内酯类抗感染药物的生物合成。在这项提案中，一个新的 基因改组技术RACHITT应用于脱硫氨基转移酶 (DES VII)来自维尼链霉菌的吡克罗霉素生物合成途径。 DES VII是一种具有非凡的固有底物灵活性的酶，我们 预计将进一步扩大其底物光谱，以包括16元环 大环内酯类。这项技术有望提供糖基转移酶 能催化12、14和16元环生物合成的酶 大环内酯类。此外，我们希望产生一个具有 对14元环大内酯那波内酯有较高的酶活性。 这将是酮内酯抗生素组合生物合成的关键一步。 它们有可能对抗耐多药细菌病原体。 建议的商业应用：不可用