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

生产像人乳低聚糖(HMOS)这样的益生菌化合物对工业生物技术来说是一个有价值的和不断扩大的市场。细菌细胞在许多过程中制造和分解低聚糖，而大肠杆菌的工程菌株可以在细胞质中合成类似人类的HMO。在优化这些产品生产的同时，产品出口显然是生产中的瓶颈。即使在像大肠杆菌这样的模型系统中，人们对细菌排出糖的了解也很少，在这个项目中，学生将识别和操纵与HMO出口有关的排出蛋白。已知的一个重要的蛋白质家族是SET外排系统，学生将寻找这个家族中更多的成员来改善这个过程的性质。同时，学生将尝试使用囊泡更详细地研究SET蛋白质的结构和功能，如果可能的话，提纯和重组蛋白质，以检查它们对HMOS和其他双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中对运输蛋白的分子微生物学和生物化学有广泛的理解，但在任何细菌系统中也基本上没有被研究。虽然该项目以约克生物学的实验室为基础，该学生是白玫瑰DTP iCASE生物经济队列的一部分，但在该项目期间，该学生也有机会与iCASE合作伙伴Glycom一起在哥本哈根工作。

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.