Directed Evolution of Glycosyltransferases

糖基转移酶的定向进化

• 批准号: 7197291
• 负责人: Kerry R Love
• 金额: $ 4.88万
• 依托单位: WHITEHEAD INSTITUTE FOR BIOMEDICAL RES
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-04-01 至 2008-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7197291
• 关键词: Active Sites; Address; Bacterial Antibiotic Resistance; Bacterial Infections; Bacteriophages; Binding; Biological Assay; Biological Factors; Carbohydrates; Chemicals; Chimeric Proteins; Clinical Trials; Condition; Dependence; Drug resistance; Enzymes; Evolution; Facility Construction Funding Category; Fellowship; Generations; Glucose; Glucosyltransferase; Glycoconjugates; Glycopeptide Antibiotics; Goals; High Pressure Liquid Chromatography; Incidence; Incubated; Individual; Infection; Investigation; Length; Libraries; Link; Love; Methods; Modification; Molecular Biology; Molecular Evolution; Names; Nature; Patients; Peptides; Phage Display; Polymerase Chain Reaction; Positioning Attribute; Production; Range; Reaction; Reaction Time; Research; Research Proposals; Solutions; Solvents; Streptavidin; Surface; System; Teicoplanin; Temperature; Testing; Therapeutic Agents; Uridine Diphosphate Glucose; Vancomycin; base; biotin 2; catalyst; chemical synthesis; crosslink; directed evolution; enzyme activity; functional group; glycosylation; glycosyltransferase; improved; interest; member; mutant; oritavancin; protein aminoacid sequence; research study; sugar; tool

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.

描述（由申请人提供）： 抗生素耐药性细菌感染发生率的增加表明需要改进结构来治疗肠球菌感染患者。对现有糖肽抗生素（例如万古霉素和替考拉宁）在糖替代品上及其周围的修饰导致了包括奥利万星在内的新疗法的临床试验。这项研究的长期目标是加深对糖基转移酶活性位点选择性的了解，以便可以鉴定新的、非天然的基于碳水化合物的构建体用于治疗耐药感染。本研究解决的具体问题是：“如何在能够糖基化非天然底物的突变糖基转移酶中修饰天然酶活性位点？”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，以快速大规模地生产糖复合物。