Interfacing Polyketide Synthase and Non-Ribosomal Peptide Synthetase Pathways Through Exploitation of Hairpin Docking Domain Pairs

通过利用发夹对接结构域对连接聚酮化合物合成酶和非核糖体肽合成酶途径

• 批准号: 2265840
• 金额: --
• 依托单位: University of Warwick
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2019
• 资助国家: 英国
• 起止时间: 2019 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2265840
• 关键词: Interfacing; Polyketide; Synthase; Non; Ribosomal

Polyketide synthases (PKSs) and non-ribosomal peptide synthetases (NRPSs) play a critical role in the biosynthesis of numerous biologically-active natural products. Acting as molecular assembly lines, these modular megasynth(et)ases construct metabolic products in a stepwise fashion from simple precursors. To maintain biosynthetic fidelity, the interactions between subunits of these enzymatic machines are mediated by docking domains (DDs); short polypeptides appended to the C- and N-termini of the subunits. This project focusses on the action of Short Linear Motif (SLiM) -hairpin docking domain (HDD) pairs, one of a handful of docking domain types known to mediate subunit interactions in hybrid NRPS-PKS systems. These docking domains are attached to the C-terminus of PKS carrier proteins and the N-terminus of their interacting NRPS domain. Recent work in the Challis group has shown these DDs are involved in the biosynthesis of HDAC inhibitors, where they are hypothesised to mediate the interaction between PKS and NRPS components of the assembly line. A range of computational, chemical and biochemical approaches will be taken to explore the prevalence of SLiM-HDD pairs in hybrid PKS-NRPSs, and how their interactions can be exploited to generate novel biosynthetic pathways. Overproduction of relevant protein domains in E.coli will form the basis for biochemical assays to assess the interaction between recombinant PKS and NRPS components containing the docking domains. Biosynthetic engineering will be undertaken to fuse the PKS and NRPS components from separate pathways, enabling the de novo production of novel HDAC inhibitors.

聚酮酶（PKS）和非核糖体肽合成酶（NRPS）在许多具有生物活性的天然产物的生物合成中起关键作用。作为分子装配线，这些模块化的大合成酶从简单的前体逐步构建代谢产物。为了保持生物合成的保真度，这些酶机器的亚基之间的相互作用是由对接结构域（DD）介导的;短多肽附加到亚基的C-和N-末端。该项目侧重于短线性基序（SLiM）-发夹对接结构域（HDD）对的作用，这是已知在混合NRPS-PKS系统中介导亚基相互作用的少数对接结构域类型之一。这些对接结构域连接到PKS载体蛋白的C-末端和它们相互作用的NRPS结构域的N-末端。查利斯小组最近的工作表明，这些DD参与HDAC抑制剂的生物合成，假设它们介导组装线的PKS和NRPS组分之间的相互作用。一系列的计算，化学和生物化学的方法将被用来探索SLiM-HDD对在混合PKS-NRPS中的流行，以及如何利用它们的相互作用来产生新的生物合成途径。大肠杆菌中相关蛋白结构域的过量产生将形成生物化学测定的基础，以评估重组PKS和含有对接结构域的NRPS组分之间的相互作用。生物合成工程将进行融合PKS和NRPS组件从不同的途径，使从头生产新的HDAC抑制剂。