Figure 5.

Purification and the activity assay of the recombinant DPYD. (A) The visualization of purified wild type and L310S mutant forms of DPYD by western blot. Lanes 1 & 4, molecular ladder (Thermo Scientific PageRuler Plus Prestained Protein Ladder); Lane 2, wild type DPYD; Lane 3, L310S mutant form of DPYD. This is an uncropped image of the gel. (B) Steady state kinetic measurement of wild type (filled circle) and L310S mutant (down filled triangle) forms of DPYD. Enzymatic activities were measured in 100 mM potassium phosphate, pH 7.0, at 50 µM NADPH. The curves were fits of data to the steady state substrate inhibition²¹.