Electrophoretic method for assessment of substrate promiscuity of a heterogeneous biocatalyst using an area imaging ultraviolet detector.
We report a new electrophoretic set-up and method for rapid specificity screening of an immobilised enzyme against a range of substrates present in a mixture. The penicillinase-catalysed reaction is carried out on-the-fly, following separation of putative substrates and preceding separation of the reaction products. The new active pixel sensor detector gives an option of using multiple detection windows on a single flow line and enables efficient on-line monitoring of this heterogeneous biocatalytic process with multiple putative substrates injected simultaneously.
Urban
Pawel L
PL
Department of Chemistry, University of York, Heslington, York, UKYO10 5DD.
Bergström
Edmund T
ET
Goodall
David M
DM
Narayanaswamy
Sreedevi
S
Bruce
Neil C
NC
D019149:Bioreactors; D002384:Catalysis; D046209:Electrophoresis, Microchip / Q000295:instrumentation / Q000379:methods*; D004798:Enzymes / Q000032:analysis*; D010405:Penicillinase / Q000378:metabolism; D014466:Ultraviolet Rays
D004798:Enzymes; D010405:Penicillinase