Improved stability of fluorinated chloramphenicol acetyltransferase by a single-isoleucine mutation

CHED 109

Man Xia Lee, ooomoodoo@gmail.com1, Natalya Voloshchuk, nvoloshc@poly.edu2, and Jin K. Montclare, jmontcla@poly.edu2. (1) Chemical and Biological Sciences, Polytechnic University, 6 Metrotech Center, Brooklyn, NY 11201, (2) Department of Chemical and Biological Sciences, Polytechnic University, 6 Metrotech Center, Brooklyn, NY 11201
In vivo incorporation of non-natural amino acids can be used to modify the complex structures and functions of proteins. Previous studies have shown an increase in thermostability of proteins by the incorporation of fluorinated amino acids in vivo. In contrast, chloramphenicol acetyltransferase (CAT) demonstrates a loss in thermostability with the incorporation of 5', 5', 5'-trifluroleucine (TFL), a leucine analog. Here, we show that a mutant form of CAT (L158I) has an increased thermostability upon the incorporation of TFL. Thus, with a single substitution of leucine to isoleucine, we have engineered a thermally stable fluorinated variant of CAT.
 

General Posters
7:30 PM-9:30 PM, Sunday, August 19, 2007 BCEC -- Exhibit Hall - B2, Poster

Division of Chemical Education

The 234th ACS National Meeting, Boston, MA, August 19-23, 2007