Synthesis of enzyme pseudo-substrates for protein engineering

ORGN 511

Birte K. Feld, Brett E. Howard, Sara K. Avrantinis, and Gregory A. Weiss. Department of Chemistry, U.C. Irvine, 516 Rowland Hall, University of California, Irvine, Irvine, CA 92697-2025
Enzyme engineering requires general methods to differentiate and detect enzyme activity amongst vast numbers of potential enzymes. To this end, we have developed two classes of enzyme pseudo-substrates that will release an indicator moiety upon reaction. This indicator will cause an easily detectable color change, which will allow us to monitor enzymatic reactions. In the first class, the highly colorgenic properties of the indoxyl functionality are utilized. We report improved conditions for the synthesis of enzyme substrates with this functionality. For example, we have tethered the indoxyl moiety to a number of different farnesyl alcohol derivatives. The second class consists of fluorogenic derivatives. The resultant molecules can provide pseudo-substrates for protein engineering applications.