Our 2023 iGEM project was born out of a need for new diverse and high throughput genetic tools for use in methanotroph engineering. We saw an opportunity to test and characterize a new library of promoters for use in methanotrophs after originally testing and reporting the efficiency of the Anderson series in e. Coli, and two kinds of methanotrophs (m. Capsulatus (bath), and Ob3B) and finding a lack of varied promoter activity in both of the methanotroph species. Our constructed library, when incorporated into an engineered methanotroph, will allow for the dynamic expression of a valuable synthetic product at a variety of constant rates that most likely will not interfere with the healthy growth of our engineered methanotrophs. In constructing and testing a new library in three separate organisms, we also hope to evaluate the ability of the new library to function, to an extent, independently of internal and external cellular factors.
Our goal is to use this dynamic promoter library to engineer the methanotroph metabolism to take harmful greenhouse gasses out of the atmosphere and create beneficial synthetic products. This approach allows us to add new technology to the field of synthetic biology and propose a mitigation strategy for detrimental greenhouse gasses in our atmosphere. Our project presents data regarding the range of activity of the Anderson series in e.coli, bath, and Ob3B while also characterizing a novel new promoter library for future use in methanotroph engineering.
We are a team of undergraduates and TAMS students from the University of North Texas working in the Henard laboratory. We are all extremely passionate about methanotrophs, cell engineering, and sustainability. We chose to compete in iGEM to take our research to the next level and to bring attention to the solution we are working on for a crucial problem facing our generation and generations to come.
As a team we have already characterized the Anderson series in M. capsulatus (bath) and e. coli and compared the dynamic range of the promoters in e. coli versus bath. Using RFP we were able to calculate relative fluorescence values for the series in both bath and e. coli using measured fluorescence and measured OD600. We have also successfully mutagenized a strong native methanotroph promoter, responsible for the production of the enzyme methane monooxygenase, to create a promoter library with varying activity in methanotrophs enabling more effective and customizable future methanotroph engineering and biocatalyst development.