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Slide LudwigLab Biocatalysis & Biosensing Research Group

Enzyme Kinetics Laboratory

Our group started with steady-state methods to study enzymatic activity by spectrophotometry and fluorimetry and expanded in the last years the methodology to study also transient kinetics. The photometers and fluorimeters are adapted with 6 or 8-cell changers, thermostats, Peltier temperature controllers and thermal incubation chambers for up to 60 cuvettes to facilitate multiple measurements. In a first round the pH dependency (pH profile, pH optima) of purified enzymes or enzyme variants are determined and stable assay conditions established. Then the determination of KM and Vmax (kcat) values follows. For special mechanistic questions our stopped-flow spectrophotometer is used. The high-throughput screening of mutagenesis libraries is performed on a plate readers using the 96-well-plate format. Oxygen-sensitive measurements can be performed in the anaerobic workstation.

Equipment
2 Perkin Elmer Lambda 35 UV/Vis Spectrophotometers
2 Agilent 8453 Diode Array UV/Vis Spectrophotometers
Hitachi U-3000 UV/Vis Spectrophotometer
Varian Cary Eclipse Fluorescence Spectrophotometer
Applied Photophysics SX20 Stopped-Flow Spectrometer
Whitley DG250 Anaerobic Workstation