Creating natural product diversity via the fabrication of a hybrid fatty acid-nonribosomal peptide synthetase

通过制造杂合脂肪酸-非核糖体肽合成酶创造天然产物多样性

• 批准号: 10339399
• 负责人: Joshua Catungal Corpuz
• 金额: $ 1.99万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-01-01 至 2022-04-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10339399
• 关键词: Achievement; Address; Amino Acids; Anabolism; Antibiotics; Antifungal Agents; Berry; Biochemistry; Biomedical Engineering; Carrier Proteins; Chemicals; Clinic; Complex; Computational Biology; Computer Models; Computing Methodologies; Coupling; Daptomycin; Data; Development; Drug Targeting; Engineering; Enzymes; Escherichia coli; Fatty Acids; Fatty-acid synthase; Future; Goals; Hybrids; Immunosuppressive Agents; In Vitro; Knowledge; Methodology; Methods; Microbe; Modeling; Modernization; Modification; Molecular; Monitor; Movement; Multiple Partners; Mutagenesis; Natural Products; Nature; Organism; Pathway interactions; Peptide Biosynthesis; Peptides; Pharmaceutical Preparations; Prevalence; Prodigiosin; Production; Productivity; Proteins; Role; Specificity; Structure; System; Therapeutic; Training; Work; X-Ray Crystallography; antimicrobial; clinically relevant; combinatorial; design; genetic technology; genome sequencing; holo-(acyl-carrier-protein) synthase; in vivo; methicillin resistant Staphylococcus aureus; microbial; novel; peptide synthase; polyketide synthase; practical application; programs; protein complex; protein protein interaction; rational design; scaffold; simulation; structural biology; success; tool; Achievement; Address; Amino Acids; Anabolism; Antibiotics; Antifungal Agents; Berry; Biochemistry; Biomedical Engineering; Carrier Proteins; Chemicals; Clinic; Complex; Computational Biology; Computer Models; Computing Methodologies; Coupling; Daptomycin; Data; Development; Drug Targeting; Engineering; Enzymes; Escherichia coli; Fatty Acids; Fatty-acid synthase; Future; Goals; Hybrids; Immunosuppressive Agents; In Vitro; Knowledge; Methodology; Methods; Microbe; Modeling; Modernization; Modification; Molecular; Monitor; Movement; Multiple Partners; Mutagenesis; Natural Products; Nature; Organism; Pathway interactions; Peptide Biosynthesis; Peptides; Pharmaceutical Preparations; Prevalence; Prodigiosin; Production; Productivity; Proteins; Role; Specificity; Structure; System; Therapeutic; Training; Work; X-Ray Crystallography; antimicrobial; clinically relevant; combinatorial; design; genetic technology; genome sequencing; holo-(acyl-carrier-protein) synthase; in vivo; methicillin resistant Staphylococcus aureus; microbial; novel; peptide synthase; polyketide synthase; practical application; programs; protein complex; protein protein interaction; rational design; scaffold; simulation; structural biology; success; tool

Abstract: Natural products and derivatives/mimics of natural products make up the majority of modern therapeutic drugs due to their unique complex organic scaffolds that contribute to their wide range of bioactivities. These natural products are synthesized by the fatty acid synthase (FAS), the nonribosomal peptide synthetase (NRPS) or the polyketide synthase (PKS). This program studies the role of protein•protein interactions in the modular synthases responsible for the formation of clinically relevant fatty acids and nonribosomal peptides for the long-term goal of re-designing and completely controlling the biosynthetic assembly lines. An example of a natural product therapeutic in the clinic is the antimicrobial daptomycin, which is a lipopeptide synthesized from a hybrid fatty acid synthase (FAS) and nonribosomal peptide synthetase (NRPS) that is used to treat methicillin- resistant Staphylococcus aureus. Thus, the hybrid FAS/NRPS are valuable systems to study and control so we may diversify natural products with new or enhanced bioactivities. Over the last three decades, true control over the biosynthetic assembly lines has remained elusive; the lack of a fundamental understanding on how the mega- synthase’s protein•protein interactions govern natural product biosynthesis has hindered this promising advance. In order to control and develop a hybrid FAS/NRPS biosynthetic pathway, first we must understand the fundamentals that guides the related biosynthetic machinery. Both FASs and NRPSs are dependent on the carrier protein (CP); the CP shuttles the growing substrate between multiple partner proteins (PP) for functionalization and incorporation into the natural product. The modular nature of fatty acid and nonribosomal peptide biosynthesis is conserved among organisms, however, the mechanism of CP•PP recognition in each pathway is distinct. Thus, to re-design these biosynthetic assembly lines, this project focuses on studying the molecular details of the CP•PP interface. We aim to 1) Characterize NRPS CP•PP interactions via X-ray crystallography, 2) establish a computational methodology to design a hybrid NRPS/FAS interaction, and 3) explore the cross-pathway productivity of the hybrid NRPS/FAS interaction. The results from these studies will be integral to the future bioengineering of these modular synthases.

摘要：天然产品和衍生产品/天然产品的模仿构成了大多数现代产品 治疗药物由于其独特的复杂有机支架而导致其广泛的生物活性范围。 这些天然产物是由非核糖体胡椒合成酶合成的 （NRP）或聚酮化合物合酶（PKS）。该程序研究蛋白质的作用•蛋白质相互作用在 负责形成临床相关的脂肪酸和非核糖体肽的模块化合酶 重新设计和完全控制生物合成组装线的长期目标。一个例子 诊所的天然产物治疗是抗菌daptomycin，它是从中合成的脂肽 杂化脂肪酸合酶（FAS）和非核糖体肽合成酶（NRPS），用于治疗甲氧西林 - 抗性金黄色葡萄球菌。这是混合FAS/NRP是研究和控制的宝贵系统，因此我们 可以通过新的或增强的生物活性使天然产品多样化。在过去的三十年中，真正控制 生物合成组装线仍然难以捉摸。缺乏对大型如何的基本了解 合酶的蛋白质•蛋白质相互作用控制天然产物生物合成，这阻碍了这一前途的进步。 为了控制和开发混合FAS/NRP的生物合成途径，首先我们必须了解 引导相关生物合成机械的基本面。 FASS和NRPS都取决于 载体蛋白（CP）; CP将多种伴侣蛋白（PP）之间的生长底物穿梭 功能化和公司为天然产品。脂肪酸和非核糖体的模块化性质 但是，肽生物合成在生物体中是保守的，但是，每种CP•PP识别的机制 途径是不同的。为了重新设计这些生物合成组装线，该项目着重于研究 CP•PP接口的分子细节。我们的目的是1）表征NRP CP•PP通过X射线相互作用 晶体学，2）建立一种设计混合NRP/FAS相互作用的计算方法，3） 探索混合NRP/FAS相互作用的跨道路生产率。这些研究的结果将 是这些模块化合酶的未来生物工程不可或缺的一部分。