Enhancing the enantioselectivity of an epoxide hydrolase by directed evolution.
[reaction: see text] The epoxide hydrolase (EH) from Aspergillus niger, which shows a selectivity factor of only E = 4.6 in the hydrolytic kinetic resolution of glycidyl phenyl ether, has been subjected to directed evolution for the purpose of enhancing enantioselectivity. After only one round of error-prone polymerase chain reaction (epPCR), enantioselectivity was more than doubled (E = 10.8). The improved mutant enzyme contains three amino acid exchanges, two of which are spatially far from the catalytically active center.
Reetz
Manfred T
MT
Max-Planck-Institut für Kohlenforschung, Kaiser-Wilhelm-Platz 1, 45470 Mülheim/Ruhr, Germany, Institut für Pharmakologie und Toxikologie, Universität Würzburg, Versbacher Strasse 9, 97078 Würzburg, Germany. reetz@mpi-muelheim.mpg.de
Torre
Claudia
C
Eipper
Andreas
A
Lohmer
Renate
R
Hermes
Marcus
M
Brunner
Birgit
B
Maichele
Andrea
A
Bocola
Marco
M
Arand
Michael
M
Cronin
Annette
A
Genzel
Yvonne
Y
Archelas
Alain
A
Furstoss
Roland
R
D001234:Aspergillus niger / Q000201:enzymology; D002384:Catalysis; D018360:Crystallography, X-Ray; D019020:Directed Molecular Evolution; D004851:Epoxide Hydrolases / Q000737:chemistry*; D007700:Kinetics; D015394:Molecular Structure; D010647:Phenyl Ethers / Q000737:chemistry; D011487:Protein Conformation; D013237:Stereoisomerism
D010647:Phenyl Ethers; C016050:phenylglycidyl ether; D004851:Epoxide Hydrolases