Gluten intolerance disorders such as Celiac sprue elicit a painful immunological response in the intestines of patients who consume gluten. Gluten is a polypeptide found in wheat, barley, and rye, and contains repeating Pro-Gln-Leu-Pro (PQLP) motifs. Currently the only treatment for gluten intolerance is elimination of gluten from the diet. Our goal is to design an enzyme therapeutic that can be taken as a drug therapeutic in pill form. The engineered enzyme will break down the immunogenic PQLP motifs before gluten reaches the small intestine and can elicit an inflammatory response. Kumamolisin-As, a protein isolated from Alicyclobacillus sendaiensis, demonstrates promising proteolysis on PQ residues, and works in the low pH of the stomach. Using the protein-engineering program “FoldIt,” we designed novel mutations to Kumamolisin to improve its activity on PQLP. Our best version of the enzyme, dubbed KumaMax, degrades gluten over 700 times faster at pH 4 than SC-PEP, an enzyme currently in clinical trials. After further protein characterization, we hope to start clinical trials on KumaMax with the future goal of creating an effective treatment for gluten intolerance.

Synthetic biology holds great promise regarding the production of important compounds, and the degradation of harmful ones. Last summer, we harnessed the power of synthetic biology to meet the world’s needs for fuel and medicine. We constructed a strain of Escherichia coli that produces a variety of alkanes, the main constituents of diesel fuel, by introducing a pair of genes recently shown convert fatty acid synthesis intermediates into alkanes. Using this method, we were able to produce on average 160 mg of alkanes per liter. Also, we identified a protease with gluten-degradation potential, and then reengineered it to have 100 times increased gluten-degrading activity, allowing for the breakdown of gluten in the digestive track when taken in pill form. Finally, to enable next-generation cloning of standard biological parts, BioBrick vectors optimized for Gibson assembly, an emerging DNA cloning technique, were constructed and used to construct the Magnetosome Toolkit: genes for biofabrication of magnetic particles.