Directed Evolution of a Glutathione Transferase for the Development of a Biosensor for Alachlor Determination

In the present work, DNA recombination of three homologous tau class glutathione transferases (GSTUs) allowed creation a library GmGSTUs. The was activity screened for identification transferase (GST) variants with enhanced catalytic towards herbicide alachlor (2-chloro-2?,6?-diethyl-N-(methoxymethyl)acetanilide). One enzyme variant (GmGSTsf) improved and binding affinity identified explored development an optical biosensor determination. Kinetics analysis molecular modeling studies revealed key mutation (Ile69Val) at subunit interface (helix ?3) that appeared to be responsible altered properties. immobilized directly on polyvinylidenefluoride membrane by crosslinking glutaraldehyde placed inner surface plastic cuvette. rate pH changes observed as result reaction followed optometrically using indicator. A calibration curve indicated linear concentration range 30–300 ?M. approach used in study can provide tools generation novel enzymes eco-efficient environment-friendly analytical technologies. addition, outcome this gives example harnessing protein symmetry design.