Understanding and exploiting bacterial sugar transporters for increased production of human prebiotics

人乳寡糖（HMOs）等益生元化合物的生产是工业生物技术一个有价值且不断扩大的市场。细菌细胞在许多过程中产生和破坏寡糖，大肠杆菌工程菌株可以在细胞质中合成类似人的HMOs。在优化这些产品生产的同时，产品出口显然是生产的瓶颈。即使在像大肠杆菌这样的模型系统中，细菌对糖的外排也知之甚少，在这个项目中，学生将识别和操纵与HMOs输出有关的外排蛋白。已知重要的蛋白质家族是Set外排系统，学生将寻找该家族中具有改进性能的其他成员用于该过程。同时，学生将尝试使用囊泡更详细地研究Set蛋白的结构和功能，并且，如果可能的话，纯化和重组蛋白质，以检查它们对HMOs和其他二糖和寡糖的亲和力和周转率。学生将在与工业直接相关的过程背景下，对转运蛋白的分子微生物学和生物化学有广泛的了解，但在任何细菌系统中也没有得到根本的研究。虽然该项目是在约克生物学的实验室进行的，学生是白玫瑰DTP iCASE生物经济队列的一部分，但在项目期间，学生也有机会在哥本哈根与iCASE合作伙伴Glycom一起工作。

The production of prebiotic compounds like human milk oligosaccharides (HMOs) is a valuable and expanding market for industrial biotechnology. Bacterial cells make and break oligosaccharides during many processes and engineered strains of the bacterium Escherichia coli can synthesize human-like HMOs in the cytoplasm. While optimizing the production of these products, it is apparent that product export is a bottleneck in production. The efflux of sugars by bacteria is poorly understood, even in model systems like E. coli, and in this project the student will identify and manipulate efflux proteins that are involved in the export of HMOs. One family of proteins known to be important are the Set efflux systems and the student will seek additional members of this family with improved properties for the process. In parallel, the student will attempt to study the structure & function of the Set proteins in more detail using vesicles, and, if possible, purify and reconstitute proteins, to examine their affinity and turnover rate for HMOs and other di- and oligo-saccharides. The student will gain a broad understanding of the molecular microbiology and biochemistry of transport proteins, in the context of a process with direct relevance to industry, but yet also fundamentally understudied in any bacterial system. While the project is based in labs at York Biology and the student forms part of the White Rose DTP iCASE Bioeconomy cohort, there are also opportunities for the student to work in Copenhagen with the iCASE partner, Glycom, during the project.