Through directed evolution, we have developed many novel FAD-dependent halogenase variants in Prof. Jared Lewis's lab at The University of Chicago. Through the CCHF, I was able to take several of these halogenases to Novartis Institute for Biomedical Research and screen these enzymes on a panel of aromatic compounds.
We were able to conduct and analyze more than 1,100 halogenation reactions while I was there. In addition to the screening we were able to accomplish, I got the chance to work in a synthetic biology lab at Novartis. In this lab, I expressed several halogenases Novartis had on hand with and without a protein chaperone, which had been shown by the Lewis lab to substantially increase yield of soluble protein for certain halogenases.
After purification, we did indeed find that this chaperone increased yields for many of the new halogenases (as high as 10-fold increases). Novartis will be sending me the genes for these halogenases soon, and I will be testing their activity on a smaller panel of substrates.
Author: Mary Andorfer