Study on amino amides and enzyme kinetics of L-asparaginase by MCE

Seven kinds of amino amides including three synthetic arylglycine amides and four normal amino amides were successfully separated by MCE with LIF detector. Using micellar electrokinetic electrophoresis, the optimized separation of the seven kinds of amino amides was achieved with FITC as the labeling reagent and polyoxyethylene lauryl ether as the surfactant in 20.0 mM borate at pH 9.2. Under the optimized conditions, linearity of L-asparagine was obtained in the range of 6.6x10(-6)-2.6x10(-4) M; the detection limit (S/N=3) was 0.7 muM. The enzyme kinetic constants of L-asparaginase using L-asparagine as substrate were also determined by this method and the kinetic parameter K(m) and V(max) for the enzymatic hydrolysis of L-asparagine were 442.0 microM and 69.9 microM/min, respectively.