FIG. 4.

Affinity chromatography HPLC of VM₁ reductase on a HiTrap Blue column (A) and purity analysis of the sample by SDS-PAGE (B). (A) VM₁ reductase activity (•). A typical elution profile with a 400-μl injection of the concentrate from step 3 was demonstrated. The gradient profile of NADPH concentration is indicated by a broken line. The protein concentration in the eluate was monitored by fluorescence (excitation, 280 nm; emission, 340 nm) (solid line). (B) Lane 1, purified enzyme (0.2 μg); lane 2, molecular mass standards. Numbers on the right indicate the size of standard proteins in kilodaltons.