Development of new tools for de novo polyketide synthase design

天然产物是多种化合物，主要由细菌和真菌产生，它们通常通过对抗竞争有机体的生长而赋予生产菌株生存优势。在这些天然产品中，多酮类药物是一个特别成功的药物类别，有40个上市实例，排在前六位的年销售额高达10亿美元。许多公司试图改善自然多样性，但收效甚微。以前用于工程聚酮合成酶(PKS)的方法可能会很慢，设计复杂，产量和产量往往很低。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合成酶(PKS)，在治疗发现和开发以及其他天然产物已经成功的领域具有广泛的应用潜力。我们的工业合作伙伴异构酶治疗公司的研究人员发现了一种突破性的技术，可以使用复合来快速生成新的和高效的PKS。我们相信，分析这些重组事件，识别潜在的重组热点，可能会为合理设计新的聚酮变性产品提供新的工具和技术。剑桥大学的研究团队将利用我们在抗生素生产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个菌株的基因组序列，其中重组的PKS已被证明产生了新的截断或延长的聚酮产品。通过对这些重排的PKS基因进行仔细的序列比较，我们的目的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之间的差异程度。这将为构建由热点边界定义的PKS基因片段的电子数据库开辟道路，这对未来从头开始设计和构建新型PKS具有潜在的价值。

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.