Team:IISER-Bhopal-India/Idea
Team MethNote To The Front!
Bringing these basic tenets into consideration, we have conceptualised a technical prototype for a robust, field-applicable methane biosensor-The MethNote. We plan to employ the soluble counterpart of the enzyme complex- Methane Monooxygenase (MMO) (used to catalyse methane to methanol conversion) isolated from the methanotrophic bacterium Methylococcus capsulatus into our chassis organism- Pichia pastoris.
Hence, the expression of soluble MMO in the methylotrophic yeast, Pichia pastoris would help integrate both these systems. In order to facilitate visualisation of the same, the P. pastoris employed would also harbour a plasmid expressing RFP under a methanol inducible promoter, 'AOX'. Effectively, we have managed to link the occurrence of RFP (Red Fluorescence Protein) fluorescence to Methane accumulation, thereby generating a methane biosensor.
The most promising aspects of our concept are its low-environmental and ecological footprint, ease of upscaling and broad-spectrum applicability.
Past Approaches
Metagenomics lab at IISER Bhopal has been working on the construction a metabolic model in M.capsulatus.(manuscript under review). This has inspired us to work with soluble counterpart of methane monooxygenase.
We would like to gratefully acknowledge the contributions, achievements, and directed efforts by past iGEM teams in addressing the burning issues of Methane / Methanol management.
The first effort dates to 2013 when Team Brazil developed a methanol biosensor in P. pastoris which aimed at detecting methanol adulteration. Subsequent efforts by Team Braunschweig’s (2014) proved to be very helpful for our project, as they were amongst the very first iGEM teams which addressed the problem of methane release in the gut of ruminants using E.coli as a chassis. They attempted to metabolize this greenhouse gas right after its production in the rumen by introducing genetic modifications like the expression of sMMO enzyme complex in E.coli.
The problems posed by Methanol and Methane were targeted by Team Oslo (2015) and Team Maryland (2016). While the former aimed to design a filter containing recombinant E. coli that would break down methanol to biomass, the latter worked on the metabolic degradation/biosequestration of methane via two pathways (Fructose and Formate) in E.coli. More recently, the high school team from Hungary (2017) highlighted the need for a methane sensor, which they attempted to develop in Methylococcus capsulatus.
We would sincerely like to acknowledge these efforts which have served as valuable references in our endeavour, and we earnestly express our gratitude towards these crucial signposts in having helped in the progress of our developing a prototype of methane biosensor.