Engineering isobutylamine N-hydroxylase for applications in antibiotic biosynthes

工程化异丁胺 N-羟化酶在抗生素生物合成中的应用

• 批准号: 8691725
• 负责人: Jessica Vey
• 金额: $ 13.11万
• 依托单位: CALIFORNIA STATE UNIVERSITY NORTHRIDGE
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-07-01 至 2016-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8691725
• 关键词: Active Sites; Address; Adverse effects; Amines; Amino Acids; Anabolism; Antibiotics; Bacterial Antibiotic Resistance; Bacterial Infections; Binding; Biochemical; Bioinformatics; Biological Factors; California; Catalysis; Climate; Communicable Diseases; Communities; Computer Simulation; Data; Development; Disease; Drug Design; Engineering; Environment; Enzymes; Faculty; Family; Family member; Flavins; Foundations; Goals; Gram-Negative Bacteria; Human; Hydroxylation; Incidence; Knowledge; Learning; Literature; Metabolic Pathway; Mixed Function Oxygenases; Multi-Drug Resistance; Mutagenesis; Mutation; One-Step dentin bonding system; Organism; Pathway interactions; Pharmaceutical Preparations; Positioning Attribute; Prevalence; Process; Production; Protein Engineering; Proteins; Research; Site-Directed Mutagenesis; Specificity; Streptomyces; Structure; Structure-Activity Relationship; Students; Substrate Specificity; Techniques; Therapeutic; Time; Universities; Work; X-Ray Crystallography; anti-cancer therapeutic; bacterial resistance; design; drug development; experience; flavin-containing monooxygenase; graduate student; infectious disease treatment; interest; member; mutant; novel therapeutics; public health relevance; skills; small molecule; structural biology; synthetic biology; tool; valanimycin

DESCRIPTION (provided by applicant): With the emergence of bacterial resistance, identification of new diseases, and the need for new therapeutics with different efficacies, our ability to design drugs to battle bacterial infections is becoming a more urgent priority. Natural products are often useful as therapeutics for humans, though problems such as side effects and production difficulties can preclude their successful development. This proposal seeks to address the need for new antibiotics by studying structure-function relationships in the valanimycin biosynthetic pathway. This naturally available antibiotic has efficacy against gram positive and gram negative bacteria, and shows some promise as an anticancer therapeutic. With this research we hope to make valanimycin amenable to a new drug development strategy, synthetic biology, in which the drug's biosynthetic pathway is engineered to allow introduction of diversity into the product. The research described here focuses on a biosynthetic step common to multiple antibiotics - flavin-dependent hydroxylation of a primary amine. The enzyme responsible for this step in the valanimycin biosynthetic pathway will be structurally and biochemically characterized. The structure activity relationships identified by those studies will be verified by bioinformatics and biochemical techniques, including mutagenesis combined with enzymatic activity and binding studies. The data yielded will enable rational design of vlmH to alter its substrate binding specificity. Such studies can be pursued on other steps of the pathway; in this way, we can introduce diversity into the valanimycin final structure. Given enough time and research, this molecule could be developed into a useful therapeutic.

描述（由申请人提供）：随着细菌耐药性的出现、新疾病的鉴定以及对具有不同功效的新疗法的需求，我们设计药物以对抗细菌感染的能力正变得更加紧迫。天然产品通常可用作人类的治疗药物，但副作用和生产困难等问题可能妨碍其成功开发。该提案旨在通过研究valanimycin生物合成途径中的结构-功能关系来解决对新抗生素的需求。这种天然可用的抗生素对革兰氏阳性和革兰氏阴性细菌有效，并显示出作为抗癌治疗的一些希望。通过这项研究，我们希望使valanimycin适合于一种新的药物开发策略，合成生物学，其中药物的生物合成途径被工程化，以允许将多样性引入产品。这里描述的研究集中在多种抗生素共同的生物合成步骤-伯胺的黄素依赖性羟基化。将在结构上和生物化学上表征在缬氨霉素生物合成途径中负责该步骤的酶。将通过生物信息学和生物化学技术，包括结合酶活性和结合研究的诱变，验证这些研究确定的结构活性关系。所得到的数据将使得vlmH的合理设计能够改变其底物结合特异性。这样的研究可以在途径的其他步骤上进行;这样，我们可以将多样性引入valanimycin的最终结构。如果有足够的时间和研究，这种分子可能会被开发成一种有用的治疗方法。