Understanding regulation of biosynthetic gene clusters to facilitate production of secondary metabolites

了解生物合成基因簇的调控以促进次级代谢产物的产生

• 批准号: 2898868
• 金额: --
• 依托单位: University of Manchester
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2023
• 资助国家: 英国
• 起止时间: 2023 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2898868
• 关键词: Understanding; regulation; biosynthetic; gene; clusters

Microbes produce a wide range of secondary metabolites with diverse medical and industrialapplications. Genes responsible for the synthesis of secondary metabolites are carried onbiosynthetic gene clusters (BGCs), often alongside transcription factors that are involved in theregulation of the BGC. The exploitation of BGCs is sometimes impossible in their native host,necessitating heterologous expression in another host, such as E. coli, to obtain good yields ofsecondary metabolites. Often, the expression of a transferred BGC is sub-optimal or occasionally itis not expressed at all. This requires modification of existing promoters within the cluster thatmust be modified to (i) be recognized by the new host's RNA polymerase; (ii) minimize interactionsbetween proteins on the BGC and those in the host; (iii) ensure the TF(s) carried on the BGCmaintain their regulatory function. For these reasons, heterologous expression currently involvesan expensive and time-consuming process of refactoring - placing each gene of the BGC under itsown promoter and then individually modifying each promoter in a semi-random manner toachieve optimal expression levels. An alternative strategy with potential for greater efficiency is topreserve the structure and regulatory context of the BGC, but to adapt it to regulatory networks ofthe new host. This rational approach requires alterations of native BGC promoters in order toachieve optimal expression in the new host.The aim of this studentship is to develop a combined experimental and computationalframework for accurate prediction and optimization of expression levels of genes within BGCsheterologously expressed in E. coli. This will enable optimizing transcriptional expression levels ofeach gene in the BGC more rapidly and with less trial-and-error than currently possible.

微生物产生多种次生代谢物，具有多种医疗和工业应用。负责次生代谢物合成的基因由生物合成基因簇 (BGC) 携带，通常与参与 BGC 调节的转录因子一起。 BGCs 在其天然宿主中的利用有时是不可能的，需要在另一个宿主（例如大肠杆菌）中进行异源表达，以获得良好的次级代谢产物产量。通常，转移的 BGC 的表达不是最佳的，或者有时根本不表达。这需要对簇内现有的启动子进行修饰，必须将其修饰为 (i) 被新宿主的 RNA 聚合酶识别； (ii) 尽量减少 BGC 上的蛋白质与宿主中的蛋白质之间的相互作用； (iii) 确保 BGC 下的 TF 维持其监管职能。由于这些原因，异源表达目前涉及昂贵且耗时的重构过程——将BGC的每个基因置于其自己的启动子下，然后以半随机的方式单独修饰每个启动子以实现最佳表达水平。另一种具有更高效率潜力的策略是保留 BGC 的结构和监管环境，但使其适应新东道主的监管网络。这种合理的方法需要改变天然 BGC 启动子，以便在新宿主中实现最佳表达。该学生的目的是开发一个组合的实验和计算框架，用于准确预测和优化在大肠杆菌中异源表达的 BGC 内基因的表达水平。这将能够比目前更快地优化 BGC 中每个基因的转录表达水平，并减少试错次数。