Team:Dalhousie/anothertest
Background
Canada’s forests represent not only an established source of economic revenue, but also a potential source for biofuel substrate. Ligno-cellulosic by-products from pulp and paper processing are removed using chemicals, heat, and water. Microbial cellulose-degrading enzymes are high-value targets for industrial applications. Focused on innovative applications of synthetic biology, the undergraduate Dalhousie iGEM team has undertaken a multi-year project to harness the degradative capacity of microorganisms to convert cellulose into ethanol for biofuel applications. Advances in DNA sequencing technology and bioinformatics have revolutionized our ability to identify useful genes in complex biological samples. We hypothesize that the porcupine microbiome, which includes microorganisms capable of digesting bark and tree resin, will be a rich source of these useful genes. If our hypothesis is correct, mining the porcupine microbiome has the advantage of finding a suite of enzymes that have evolved to work in concert to efficiently degrade cellulose.
The Project
Using a combination of metagenomic library construction and bioinformatic analysis we aim to find known cellulolytic enzymes as well as discover novel enzymes. Screening of our metagenomic library will allow us to look for a diverse selection of enzymes, including cellulolytic and lignolytic ones by plating on cellulose-only media. The clones that grow can be sequenced for confirmation and cloned into biobricks. Our new bioinformatic pipeline now allows us to look for conserved domains rather than known enzymes. This is extremely powerful as it may allow us to discover new enzymes that share catalytic domains with cellulases but are otherwise totally novel. The goal of this project is to construct a bioreactor where cellulose would be the input, D-glucose the output and E.coli expressing a suite of cellulolytic enzymes would be the workhorse, converting one to the other. Future goals would be to combine this in co-culture with yeast for the last step in bio-ethanol production.
