Short Summary: What We Did
In this project we produced synthetic enzymes to remove red wine stains from fabric. These enzymes are designed to replace perchloroethylene (PERC), a toxic solvent used in dry cleaning that will soon be banned in France.
We listened to people who will use our product, conducting face-to-face interviews with every single dry cleaner in Paris. From this came a plan for a realistic product, a stain fighting enzymatic pretreatment compatible with existing cleaning technologies and workflows.
We isolated microbes capable of degrading red wine from a library of 186 strains taken from vineyard soil samples collected around the world with the help of our fellow iGEMers. The most effective microbes were submitted for whole genome resequencing, then analyzed to produce a short list of candidate stain-fighting enzymes.
We modeled enzymatic activity at the fabric surface and determined that activity could be substantially improved if the enzymes had a moderate binding affinity for the fabric itself. This effectively increases the enzyme concentration at the fabric surface and reduces the quantity of enzymes lost through diffusion into the medium.
We identified short peptides with affinity to cotton, linen, wool, polyester and silk using the method of phage display. The resulting Fabric Binding Domains (FBDs) were quantitatively characterized using ELISA, to determine peptides with optimal affinity as determined by our model.
We developed a high-throughput assay to quantify stain removal on real cloth. Laser-cut fabric samples are sealed to 96-well microplates and imaged on flat-bed scanners. We coded custom image analysis software to identify each circular fabric sample and measure the stain intensity.
We constructed new BioBricks, fusions of our most promising fabric binding domains to our favorite wine-degrading enzymes. The resulting proteins were expressed, purified and characterized both in vitro (in solution) and in situ (on real stained fabric).
We hope these pages convince you that our project is mature, thoroughly documented and effective. We believe that product is ready solve one small problem that real people have every day.