Engineered probiotics are the next-generation biotherapeutics. However, there are no reasonable solutions to several problems, such as colonization failure, insufficient population density, and irregular expression of exogenous genes. These shortages hinder the spread and depth of application of engineered probiotic therapy.
More on the problem
With the increasing focus on the harm of a high-sugar diet, numerous people intend to consume sugar substitutes rather than sugar itself. Erythritol stands out from the crowd of traditional sugar substitutes due to its excellent security and complete zero heat.
We designed an engineered probiotic platform based on erythritol. The engineered probiotics' colonization duration and population density got enhanced, and the curing genes were induced with the erythritol-adding diet.
Meanwhile, we gave our engineered bacteria more applications, building a multi-functional platform that benefits human health and cures specific diseases.
More on the design
We assayed the function of the erythritol utilizing pathway into E. coli, and tested the growth state under different carbon source conditions.
More on the results
We created a hybrid promoter pJ23101-eryO, which could be responsed by ery regular protein EryD. Then we tested the function of erythritol based induction system in E. coli.
More on the results
We hope to use erythritol as substrate producing curing compound L-erythrose.
We tested the PQQ production in E. coli DH5alpha and E coli Nissle 1917 introducing pqq cassette.
We used thrombus curing gene tmadh to explore our further platform application.
Based on the theory that structure determines function, we obtained the best cascade sequence of multi-enzyme cascade by protein structure prediction, protein structure align and protein docking with small molecules. At the same time, the validity of the gene sequence of eryEFG transport system is verified by modeling, and we hope it can be used in subsequent experiments.
More on the Protein Modeling
As a probiotic, the low colonisation of EcN often affects its function. By introducing erythritol utilizing pathway, we try to improve its colonization capacity. Based on GEM model and enzyme kinetic model, we simulate the growth rate of EcN as well as the synthesis of its products. Further, we apply dFBA co-culture algorithm framework to go for a more accurate simulation of EcN colonisation in the gut. Our modelling results are validated by well-established algorithms.
More on the Metabolic ModelingBeginning with a trip to traditional sugar culture, we outreached enterprises on the erythritol industry chain and talked with experts to get feedback on our project. Also, we explore the possibility of commercializing the subject and bringing our understanding of life to the public.
More on the Human Practices