查看文章

Formate dehydrogenase (FDH, EC 1.2.1.2) was purified from Candida boidinii cells in a single step by biomimetic‐dye affinity chromatography. For this purpose, seven' biomimetic analogues of the monochlorotriazine dye, Cibacron® Blue 3GA (CB3GA), and parent dichloro‐triazine dye, Vilmafix^® Blue A‐R (VBAR), bearing a car‐boxylated structure as their terminal biomimetic moiety, were immobilized on crosslinked agarose gel, Ultrogel^® A6R. The corresponding new biomimetic‐dye adsorbents, along with nonbiomimetic adsorbents bearing CB3GA and VBAR, were evaluated for their ability to purify FDH from extracts obtained after press‐disintegration of C. boidinii cells. Optimal conditions for maximizing specific activity of FDH in starting extracts (1.8 U/mg) were realized when cell growth was performed on 4% methanol, and press disintegration proceeded in four consecutive passages before the homogenate …