Collaborative Research: RUI: Elucidating Design Rules for non-NRPS Incorporation of Amino Acids on Polyketide Scaffolds

合作研究：RUI：阐明聚酮化合物支架上非 NRPS 氨基酸掺入的设计规则

• 批准号: 2300891
• 负责人: Shaun McKinnie
• 金额: $ 32.48万
• 依托单位: University of California-Santa Cruz
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-15 至 2027-03-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2300891&HistoricalAwards=false
• 关键词: Collaborative; Research; RUI; Elucidating; Design; Collaborative; Research; RUI; Elucidating; Design

More bacteria have become resistant to current antibiotics. Natural compounds with antibiotic properties are assembled in a modular fashion inside cells. The modular nature allows for the design of hybrid pathways to produce non-natural molecules. The mechanisms of individual bond-forming reactions in an antibiotic molecule will be investigated. The intention is to develop a degree of insight that would enable the shuffling of building blocks to produce novel antibiotics. The concept is akin to molecular building blocks. In addition, this work will provide a foundation for training undergraduate students in conducting biotechnological research and communicating science to their community. The project will attempt to uncover novel enzymatic mechanisms that facilitate production of unique natural products. Specifically, this work will elucidate mechanistic underpinnings of amino acid incorporation on polyketide scaffolds. The project focuses on the model molecule TLN-05220, which is an Echinosporamicin-type antibiotic with antibacterial activity against several strains of methicillin-resistant Staphylococcus aureus and vancomycin-resistant Enterococci (VRE), and cytotoxic activities against several human tumor cell lines. Individual gene functions in TLN-05220 biosynthesis will be identified using gene knockouts in the wild-type producing organism. Potential substrate binding sites and catalytic residues of amino acid-incorporating enzymes will be identified using homology modeling. Expression and characterization of individual enzymes will elucidate mechanism(s) of amino acid incorporation into natural product structures. The minimally necessary genes to produce TLN-05220 will be cloned and expressed in a heterologous host.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

更多的细菌已经对当前的抗生素具有抗药性。具有抗生素特性的天然化合物以模块化的方式组装。模块化的性质允许设计杂交途径以产生非天然分子。将研究抗生素分子中个体形成反应的机制。目的是开发一定程度的见解，使构建基块的改组能够产生新颖的抗生素。 该概念类似于分子构建块。 此外，这项工作将为培训本科生进行生物技术研究并向社区传达科学的基础。该项目将试图发现新型的酶促机制，以促进产生独特的天然产品。具体而言，这项工作将阐明在聚酮化合物支架上氨基酸掺入的机械基础。该项目侧重于模型分子TLN-05220，该项目是一种具有抗菌活性的棘孢子蛋白型抗生素，可抗抗甲氧西林抗甲氧西林的金黄色葡萄球菌和万古霉素的葡萄球菌葡萄球菌和抗Vancomycin的抗抗菌素（VRE）（VRE）（VRE）（VRE）（VRE），以及抗细胞毒素的活性。 TLN-05220生物合成中的单个基因功能将使用野生型生成生物中的基因敲除鉴定。 将使用同源性建模鉴定出氨基酸酶的氨基酸结合位点和催化残基。 单个酶的表达和表征将阐明氨基酸掺入自然产物结构中的机制。生产TLN-05220的最小基因将在异源主机中克隆并表达。该奖项反映了NSF的法定任务，并被认为是值得通过基金会的知识分子优点和更广泛影响的评估标准来通过评估来支持的。