Development of new tools for de novo polyketide synthase design

开发用于从头设计聚酮合酶的新工具

• 批准号: BB/M012158/1
• 负责人: Peter Leadlay
• 金额: $ 8.64万
• 依托单位: University of Cambridge
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2015
• 资助国家: 英国
• 起止时间: 2015 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FM012158%2F1
• 关键词: Development; new; tools; de; novo

Natural products are diverse chemical compounds, produced chiefly by bacteria and fungi, that confer a survival advantageon the producing strain often by antagonising the growth of competing organisms. Among such natural products,polyketides are a particularly successful drug class, with >40 marketed examples, the top six with peak annual sales of >$1billion. Many companies have tried to improve upon natural diversity, with limited success. Previous methods used toengineer polyketide synthases (PKS), the proteins which generate these products, can be slow, are complex to design, andhave frequently poorly productive and yielding.We aim to develop new synthetic biology tools for de novo syntheticgeneration of productive polyketide synthases (PKS), with broad potential for application in therapeutic discovery anddevelopment and in other areas where natural products have been successful.Researchers at our industrial partner Isomerase Therapeutics have discovered a groundbreaking technique for usingrecombination to rapidly generate novel and productive PKS. We believe that analysing these recombination events andidentifying potential recombination hotspots could lead to new tools and techniques for rational design of new polyketidenatural products. The Cambridge research team will use our extensive expertise in genome sequencing and analysis ofantibiotic-producing bacteria and collaborate with Isomerase to obtain genome sequence of up to 60 strains in whichrecombined PKS have been shown to give rise to novel truncated or elongated polyketide products. By careful sequencecomparisons between such rearranged PKS genes, we aim to establish the identity of those 'hotspot' regions in whichrecombination favours a successful outcome; and to establish the extent to which these hotspot regions vary betweendifferent PKS. This would open the way to the construction of an in silico database of PKS gene fragments defined byhotspot boundaries, a potentially valuable asset in the future design and construction of novel PKS from scratch.

天然产物是多种化学化合物，主要由细菌和真菌产生，通常通过对抗竞争生物体的生长来赋予生产菌株生存权。在这些天然产品中，聚酮化合物是一个特别成功的药物类别，有超过40个上市的例子，前六名的年销售额超过10亿美元。许多公司试图改善自然多样性，但成效有限。以往用于工程化聚酮酶（PKS）（产生这些产物的蛋白质）的方法可能是缓慢的，设计复杂的，并且经常具有低生产率和产量。我们的目标是开发新的合成生物学工具用于从头合成产生生产性聚酮酶（PKS），在治疗发现和开发以及其他天然产物已经取得成功的领域具有广泛的应用潜力。研究人员在我们的工业合作伙伴Isomerase Therapeutics，我们发现了一种突破性的技术，可以使用重组来快速产生新的和多产的PKS。我们相信，分析这些重组事件和识别潜在的重组热点可能会导致新的工具和技术，合理设计新的聚酮变性产品。剑桥研究小组将利用我们在产酶细菌基因组测序和分析方面的广泛专业知识，与异构酶合作，获得多达60种菌株的基因组序列，其中重组PKS已被证明能产生新型截短或延长的聚酮产物。通过对重排的PKS基因进行仔细的序列比较，我们的目标是确定那些重组有利于成功结果的“热点”区域的身份，并确定这些热点区域在不同PKS之间的变化程度。这将为构建由热点边界定义的PKS基因片段的计算机数据库开辟道路，这是未来从头开始设计和构建新型PKS的潜在宝贵资产。

项目成果

Sulfation and amidinohydrolysis in the biosynthesis of giant linear polyenes.

• DOI: 10.3762/bjoc.13.238
• 发表时间: 2017
• 期刊: Beilstein journal of organic chemistry
• 影响因子: 2.7
• 作者: Hong H;Samborskyy M;Usachova K;Schnatz K;Leadlay PF
• 通讯作者: Leadlay PF

Diversity oriented biosynthesis via accelerated evolution of modular gene clusters.

通过模块化基因簇的加速进化进行面向多样性的生物合成。

• DOI: 10.17863/cam.18018
• 发表时间: 2017
• 作者: Wlodek A