Combinatorial Biosynthesis of Polyketides

聚酮化合物的组合生物合成

• 批准号: 6999390
• 负责人: DANIEL V. SANTI
• 金额: $ 10万
• 依托单位: KOSAN BIOSCIENCES, INC.
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-01 至 2006-03-05
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6999390
• 关键词: biological products; biotechnology; combinatorial chemistry; high throughput technology; liquid chromatography mass spectrometry; nuclear magnetic resonance spectroscopy; peptide chemical synthesis; polyketide synthase; synthetic protein

DESCRIPTION (provided by applicant): Polyketides are complex natural products that are abundant in soil bacteria. From fewer than 10,000 known polyketides, a large number of therapeutic agents have been derived, covering every important therapeutic area. Complex polyketides are synthesized by polyketide synthase (PKS) consisting of sets (modules) of catalytic domains acting sequentially to build long carbon chains. Rudimentary knowledge of the rules governing the ability of PKS modules or domains to interact productively exists but the level of understanding has negated many attempts to engineer desired polyketides. The long-term aim of this proposal is to overcome this problem and to determining how to construct almost any PKS, and thus any polyketide, by combining heterologous modules in a productive fashion. Its novelty lies in the analysis of hundreds or thousands of module combinations in a semi-empirical approach and selecting those that work well. This is enabled by our recent development of high-throughput long gene synthesis technology, and a design of generic PKS modules that each contains the same restriction sites at module and domain boundaries, enabling a "building block" approach to combinatorial biosynthesis. The knowledge base for rational PKS engineering will bring enormous benefits to the manufacture of desired therapeutics as well as the availability of fermentation-derived structures to be used as intermediates in chemical synthesis and manufacturing. The specific aims of this Phase I proposal are (1) to test functionality of 2-module hybrid PKS systems with >200 combinations and identify those that produce polyketide product and (2) to test the connectivity of the results obtained with overlapping modules in the biomodular system to rationally create larger polyketide chains. The resulting technology will be expanded and perfected in Phase II research for the production of natural products derived from plant, animal, and microbial sources, such as antibiotics, anticancer drugs, and other therapeutic agents, and useful synthetic starting materials for manufacturing polyketides of current interest.

描述（由申请人提供）：聚酮化合物是土壤细菌丰富的复杂天然产物。从不到10,000个已知的聚酮化合物中，已经得出了大量的治疗剂，涵盖了每个重要的治疗区域。复杂的聚酮化合物由由聚酮化合物合酶（PK）合成，由催化域的集合（模块）组成，依次作用以构建长碳链。对规则的基本知识，该规则管理PKS模块或领域有效地存在相互作用的能力，但理解水平否定了许多试图设计了所需的聚酮化合物的尝试。该提案的长期目的是克服这个问题，并通过以生产性的方式组合异源模块来确定几乎所有PK，因此，因此都可以构建任何PK。它的新颖性在于以半经验的方法对数百或数千个模块组合进行分析，并选择效果很好的模块组合。我们最近开发了高通量长基因合成技术以及每个通用PKS模块的设计来实现这一点，每个模块都包含模块和域边界处相同的限制位点，从而实现了组合生物合成的“构建障碍”方法。理性PKS工程的知识库将为所需的治疗剂的生产带来巨大的好处，以及用于化学合成和制造中中间体的发酵衍生结构的可用性。该阶段I提案的具体目的是（1）测试具有> 200个组合的2模块混合PKS系统的功能，并识别生产聚酮化合物的产品和（2）测试用生物调节系统中重叠模块获得的结果的连通性，以便于理性地产生较大的聚酮链。在II期研究中将扩展和完善所得技术，以生产源自植物，动物和微生物来源的天然产品，例如抗生素，抗癌药物以及其他治疗剂，以及用于生产当前利益的多吉碱的有用合成起始材料。