Improving Glycosyltransferase by RACHITT Gene Shuffling

描述（由申请人提供）：参与连接的糖基转移酶 聚酮大环内酯与脱氧糖是关键酶的组合 新的大环内酯类抗感染剂的生物合成。在这项提案中，一个新的 将基因改组技术RACHITT应用于脱乙酰氨基转移酶， (Des VII）来自委内瑞拉链霉菌的吡克罗霉素生物合成途径。 Des VII是一种具有显着的固有底物灵活性的酶，我们 期望进一步扩展其底物谱以包括16元环 大环内酯。这项技术有望提供糖基转移酶 可以催化12-、14-和16-元环的生物合成的酶 大环内酯类。此外，我们期望产生具有以下特性的重组DesVII： 对14-元环大环内酯那波内酯具有较高的酶活性。 这将是组合生物合成酮内酯类抗生素的关键步骤 有可能对抗多重耐药细菌病原体。 拟议商业应用：不可用

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