Directed Evolution of Glycosyltransferases

描述（由申请人提供）： 抗生素抗性细菌感染的发病率增加表明需要改进的构建体来治疗肠球菌感染的患者。对现有糖肽类抗生素（如万古霉素和替考拉宁）在糖替代物上和周围的修饰导致了包括奥利万星在内的新治疗方法的临床试验。这项研究的长期目标是提高对糖基转移酶活性位点选择性的理解，以便可以鉴定新的非天然碳水化合物构建体用于治疗耐药感染。在本研究的背景下解决的具体问题是：“在能够使非天然底物糖基化的突变糖基转移酶中，天然酶活性位点是如何修饰的？“这项研究将开发一种现有的分子生物学工具，噬菌体展示，用于突变糖基转移酶的分子进化。我们相信，鉴定能够糖基化新底物的酶将继续产生能够与化学合成竞争的催化剂，以快速和大规模生产糖缀合物。

DESCRIPTION (provided by applicant): The increasing incidence of antibiotic resistant bacterial infections indicates the need for improved constructs to treat enterococcal infected patients. Modification of existing glycopeptide antibiotics, such as vancomycin and teicoplanin, on and around the sugar substitutents has led to the clinical trials of new treatments, including oritavancin. The long-term goal of this research is to develop an improved understanding of the active-site selectivity of glycosyltransferases so that new, non-native carbohydrate-based constructs may be identified for treatment of drug-resistant infections. The specific question addressed in the context of this research is: "How is the native enzyme active site modified in a mutant glycosyltransferase that is capable of glycosylating a non-native substrate?" This research will develop an existing tool in molecular biology, phage display, for the molecular evolution of mutant glycosyltransferases. We believe that identifying enzymes that can glycosylate new substrates will segue into the generation of catalysts able to compete with chemical synthesis for the rapid and large scale production of glycoconjugates.