Directed Evolution of Glycosyltransferases

描述(由申请人提供)： 越来越多的耐药细菌感染表明需要改进的结构来治疗肠球菌感染的患者。对现有的糖肽抗生素，如万古霉素和替考拉宁，在糖替代品上和周围进行修饰，已经导致了包括奥立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更好地了解糖基转移酶的活性部位选择性，以便找到新的、非天然碳水化合物为基础的结构来治疗耐药感染。这项研究的具体问题是：“在突变的糖基转移酶中，能够糖基化非天然底物的天然酶活性部位是如何修饰的？”这项研究将开发分子生物学中现有的工具-噬菌体展示，用于突变糖基转移酶的分子进化。我们相信，识别能够糖基化新底物的酶将有助于产生能够与化学合成竞争的催化剂，以快速和大规模地生产糖结合物。

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.