Gorbach Z V, Maglysh S S, Konovalenko O V
Ukr Biokhim Zh (1978). 1984 Mar-Apr;56(2):175-8.
Lactate dehydrogenase M4-isoform in the homogeneous state was isolated from the rat liver by successive application of sulphate-ammonium fractionation, phosphocellulose ion-exchange chromatography with high-affinity elution of 1 mM NADH and subsequent hydroxyl apatite fractionation. The method permits obtaining the preparation amounts of the enzymic protein with yield 37.5%, specific activity 386.8 units per 1 mg of protein. It is established that 1 mM NAD+, 10 mM pyruvate and 100 mM lactate are also effective as agents of the selective enzyme elution.
通过依次应用硫酸铵分级分离、用1 mM NADH进行高亲和力洗脱的磷酸纤维素离子交换色谱法以及随后的羟基磷灰石分级分离,从大鼠肝脏中分离出均一状态的乳酸脱氢酶M4同工型。该方法能够获得酶蛋白的制备量,产率为37.5%,比活性为每1 mg蛋白质386.8单位。已确定1 mM NAD +、10 mM丙酮酸和100 mM乳酸作为选择性酶洗脱剂也有效。