Probing the Substrate Specificity of Polyketide Synthase

聚酮合酶的底物特异性探讨

• 批准号: 6445777
• 负责人: CLAY CC WANG
• 金额: $ 3.66万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-02-04 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6445777
• 关键词: Escherichia coli; bioengineering /biomedical engineering; enzyme mechanism; enzyme substrate; enzyme substrate complex; polyketide synthase; postdoctoral investigator; protein purification

DESCRIPTION: (provided by applicant) Polyketides are a large family of structurally diverse and complex natural products with a wide range of biological functions. In nature, polyketides are biosynthesized by large polyketide synthase enzymes (PKS) through the successive condensations of simple monomers. The long-term goal of the proposed research is to understand the PKSs in order to generate novel polyketides through bioengineering techniques. For modular PKS, each individual module contains all the necessary active sites to carry on one round of synthesis. The proposed research will express and purify the individual modules of 6 deoxyerythronolide B synthase (DEBS). DEBS is a modular PKS responsible for the biosynthesis of the antibiotic erythromycin. Substrates are designed to specifically probe the underlying rules for binding and processing of different substrates by the individual modules. Native and hybrid bimodular constructs will also be prepared and subjected to the novel substrates to understand the incorporation of the substrates by the first module and transfer to the second module for further chemical transformations. Understanding the substrate specificities of the PKS is an important step towards combinatorial biosynthesis of diverse polyketides.

描述：（由申请人提供）聚酮化合物是一个大家族， 结构多样和复杂的天然产品， 生物功能。在自然界中，聚酮化合物是通过大规模生物合成的。 聚酮合酶（PKS）通过连续的缩合， 简单单体这项研究的长期目标是了解 为了通过生物工程产生新的聚酮化合物， 技术.对于模块化PKS，每个单独的模块包含所有必要的 活性位点进行一轮合成。拟议的研究将 表达和纯化6-脱氧腺苷酸B合酶的各个模块 （DEBS）。DEBS是一个模块化PKS，负责生物合成的 抗生素红霉素基底被设计成专门探测 粘合和处理不同基材的基本规则 单独的模块。天然和杂交双模构建体也将被 制备并经受新型底物以理解掺入 然后传送到第二模块， 进一步的化学转化。了解底物特异性 PKS是朝向多种生物组合生物合成重要步骤 聚酮化合物。