Engineering Novel Allosteric Transcription Factor Recognition: Applications in Bio-catalysis and Synthetic Biology

本项目的目的是探索具有新的变构识别能力的工程转录因子(Tf)的潜力，并将这些新的Tf应用于高通量选择新的生物催化剂。TFS可用于一系列应用，例如，筛选生物质有价的酶过程[1]，通过适应性实验室进化进化新的酶和菌株[2]，以及检测环境污染物和生物修复。该项目将致力于改造Marr型TFS的识别特性，以识别天然和合成来源的替代芳香族分子[1]，然后应用这些新的识别能力来进化出具有新底物耐受性和催化性能特征的酶。这些新的酶和细胞功能将为一些社会经济驱动因素和环境需求提供潜在的解决方案，例如全球可持续发展的文化压力，并为生产精细化工产品和中间体提供替代路线。曼彻斯特生物技术研究所的博士生将由Neil Dixon博士、David Leys教授和Nick Turner共同指导，重点将集中在i)工程Marr型转录因子以识别具有环境和工业重要性的非天然芳香化合物，ii)应用工程TF进化新的酶功能和优化生物催化能力，iii)将这些单独的酶和TF整合到整个细胞生物转化的生物合成途径中，使用合成生物学工具和原理设计开发的途径。学生将在生物技术、微生物基因表达调控、生物催化、定向进化、微生物发酵、分子生物学和生物分析方法(如GC-MS)的广泛方面进行培训。该项目将适合对未来生物技术、生物催化和生物处理职业感兴趣的个人。

The aim of this project is to explore the potential of engineering transcription factors (TF) with novel allosteric recognition capabilities, and then to apply these novel TFs in the high-throughput selection of new biocatalysts. TFs can be used for a range of applications for example; screening enzymatic processes for biomass valorisation [1], evolving novel enzymes and strains via Adaptive Laboratory Evolution [2], and detection of environmental pollutants and bio-remediation. This project will seek to engineer the recognition properties of MarR-type TFs to recognise alternative aromatic molecules of both natural and synthetic origin [1], and then apply these novel recognition capabilities to evolve enzymes with new substrate tolerance and catalytic performance characteristics. These novel enzymatic and cellular functionalities will provide potential solutions to a number of socioeconomic drivers and environmental demands such as cultural pressure for global sustainability and provide alternative routes for the production of fine-chemicals products and intermediates.The PhD student based in Manchester Institute of Biotechnology will be co-supervised by Dr. Neil Dixon, and Profs David Leys, and Nick Turner, and will be focused on i) engineering MarR-type transcriptional factors to recognise a non-natural aromatic chemicals of environmental and industrial importance, ii) applying the engineered TFs to evolve novel enzymatic functionality and optimise bio-catalytic capability, iii) integrating these individual enzymes and TFs into biosynthetic pathways for whole-cell biotransformation, and iv) using synthetic biology tools and principles to engineer the developed pathway.The student will be trained in broad aspects of biotechnology, microbial gene expression regulation, biocatalysis, directed evolution, microbial fermentation, molecular biology and bio-analytical methods such GC-MS. This project would suit individuals interested in future careers in biotechnology, biocatalysis and bioprocessing.