Fluorinated chloramphenicol acetyltransferase thermostability and activity profile: improved thermostability by a single-isoleucine mutant.
A lysate-based thermostability and activity profile is described for chloramphenicol acetyltransferase (CAT) expressed in trifluoroleucine, T (CAT T). CAT and 13 single-isoleucine CAT mutants were expressed in medium supplemented with T and assayed for thermostability on cell lysates. Although fluorinated mutants, L82I T and L208I T, showed losses in thermostability, the L158I T fluorinated mutant demonstrated an enhanced thermostability relative to CAT T. Further characterization of L158I T suggested that T at position 158 contributed to a portion of the observed loss in thermostability upon global fluorination.
Voloshchuk
Natalya
N
Department of Chemical and Biological Sciences, Polytechnic University, 6 Metrotech Center, Brooklyn, NY 11201, USA.
Lee
Man Xia
MX
Zhu
Wan Wen
WW
Tanrikulu
Ismet Caglar
IC
Montclare
Jin Kim
JK
D015500:Chloramphenicol O-Acetyltransferase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D004795:Enzyme Stability; D005461:Fluorine / Q000737:chemistry*; D007532:Isoleucine / Q000235:genetics*; D008958:Models, Molecular; D009154:Mutation; D013696:Temperature
D007532:Isoleucine; D005461:Fluorine; D015500:Chloramphenicol O-Acetyltransferase