Biosynthesis of nucleoside antibiotics targeting bacterial translocase

针对细菌转位酶的核苷抗生素的生物合成

• 批准号: 8281422
• 负责人: Steven Gary Van Lanen
• 金额: $ 42.99万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-06-15 至 2015-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8281422
• 关键词: 3-hydroxybutanal; Acquired Immunodeficiency Syndrome; Amino Acids; Anabolism; Animals; Anti-Bacterial Agents; Antibiotics; Bacteria; Biochemical; Biochemical Reaction; Biological Assay; Biological Factors; Carbon; Case Study; Cell Wall; Cessation of life; Clinic; DNA Fingerprinting; Development; Disease; Drug resistance; Drug resistance in tuberculosis; Enzymes; Family; Fluorescence; Gene Silencing; Generations; Genes; Genetic; Genotype; Glycine Hydroxymethyltransferase; Goals; Health; Hexoses; Hospitals; Human; In Vitro; Infection; Libraries; Modeling; Modification; Multi-Drug Resistance; Natural Product Drug; Nucleosides; Parents; Peptidoglycan; Pharmaceutical Preparations; Physical condensation; Property; Public Health; Reaction; Recombinant Proteins; Research; Streptomyces; Structure-Activity Relationship; System; Testing; Tuberculosis; United States; base; biomaterial compatibility; combinatorial; community setting; design; drug discovery; expectation; improved; in vivo; inhibitor/antagonist; killings; methicillin resistant Staphylococcus aureus; novel; pathogen; scaffold; sugar; tool; translocase

DESCRIPTION (provided by applicant): The emergence of multiple drug resistant pathogens is becoming problematic worldwide, and the discovery of new antibiotics continues to decline. The broad, long-term objective of this proposal is to discover, characterize, and develop a new structural class of antibiotics-nucleoside antibiotics-that target bacterial translocase I involved in peptidoglycan cell wall biosynthesis. The specific aims of this proposal are (I) to characterize the assembly and incorporation of the aminoribosyl moiety found within the family of lipopeptidyl-nucleoside antibiotics and (II) to functionally and mechanistically characterize enzymes catalyzing novel or unusual biochemical reactions represented herein by a new family of serine hydroxymethyltransferase-like enzymes that are hypothesized to catalyze an aldol-type condensation to form unusual nonproteinogenic amino acids. Specific Aim I and II will be achieved by using the robust genetic system developed for the lipopeptidyl nucleoside A-90289-producing strain for in vivo studies employing gene inactivation and cross-complementation, and recombinant proteins will be exploited for functional and mechanistic studies in vitro. The results will establish a new mechanism for incorporating ribosyl units into natural product scaffolds and will establish a paradigm for the entry into high-carbon nucleoside antibiotics. The results will be essential for our long-term goals of searching for new nucleoside antibiotics using genetic information (a genotype-to-chemotype approach), will allow for the structural diversification of the parent scaffolds using combinatorial biosynthesis and total synthesis, and will provide the basis for the design of second generation antibiotics with improved biocompatibility and pharmacological properties.

描述（由申请人提供）：多种耐药病原体的出现正在成为世界范围内的问题，并且新抗生素的发现持续下降。该提案的广泛、长期目标是发现、表征和开发一种新结构的抗生素（核苷抗生素），其靶向参与肽聚糖细胞壁生物合成的细菌易位酶 I。该提案的具体目标是（I）表征脂肽基核苷抗生素家族中发现的氨基核糖基部分的组装和掺入，以及（II）功能和机制上表征催化新颖或不寻常的生化反应的酶，本文中以新的方法为代表。丝氨酸羟甲基转移酶样酶家族，推测其催化羟醛型缩合形成不寻常的非蛋白氨基酸。具体目标 I 和 II 将通过使用为脂肽核苷 A-90289 生产菌株开发的强大遗传系统来实现，用于采用基因失活和交叉互补的体内研究，并且重组蛋白将用于体外功能和机制研究。研究结果将建立一种将核糖基单元纳入天然产物支架的新机制，并将为进入高碳核苷抗生素建立范例。这些结果对于我们使用遗传信息（基因型到化学型方法）寻找新核苷抗生素的长期目标至关重要，将允许使用组合生物合成和全合成实现母体支架的结构多样化，并将提供为设计具有改善的生物相容性和药理学特性的第二代抗生素奠定了基础。