Pigiet V P, Conley R R
J Biol Chem. 1977 Sep 25;252(18):6367-72.
A scheme is described for the large scale purification of thioredoxin, thioredoxin reductase, and glutathione reductase. The scheme is based on an initial separation of thioredoxin from the two reductases by affinity chromatography on agarose-bound N6-(6-aminohexyl)-adenosine 2',5'-bisphosphate (agarose-2',5'-ADP). The two reductases were then separated by hydrophobic chromatography and purified separately to homogeneity. Thioredoxin was purified to homogeneity by immunoadsorption to agarose containing immobilized goat anti-thioredoxin. Overall yields for thioredoxin, thioredoxin reductase, and glutathione reductase exceeded 80% in each case. Both reductases exhibit an absorption band at approximately 320 nm which appears due to a residual amount of tightly bound NADP. Presence of this absorption band has no apparent effect on the specific activity of either enzyme.
本文描述了一种用于大规模纯化硫氧还蛋白、硫氧还蛋白还原酶和谷胱甘肽还原酶的方案。该方案基于通过琼脂糖结合的N6-(6-氨基己基)-腺苷2',5'-二磷酸（琼脂糖-2',5'-ADP）亲和色谱法，首先将硫氧还蛋白与两种还原酶分离。然后通过疏水色谱法将两种还原酶分离，并分别纯化至均一性。硫氧还蛋白通过与含有固定化山羊抗硫氧还蛋白的琼脂糖进行免疫吸附纯化至均一性。硫氧还蛋白、硫氧还蛋白还原酶和谷胱甘肽还原酶的总产率在每种情况下均超过80%。两种还原酶在约320nm处均显示出一条吸收带，这是由于残留的紧密结合的NADP所致。该吸收带的存在对任何一种酶的比活性均无明显影响。
