Bagdonas A S, Sabaliauskene V L, Iuodka B A
Biokhimiia. 1981 May;46(5):802-8.
A new method for isolation of polynucleotide phosphorylase from E. coli, including ion-exchange chromatography and gel-filtration has been developed. The method results in 300-fold purification of the enzyme, which being devoid of nuclease and phosphatase activities can further be utilized for oligonucleotide synthesis. It was shown that upon storage the enzyme loses the primer-independent activity and in the absence of NaCl can be used for further syntheses. An addition of NaCl stimulates the elongation of the oligonucleotide chain. Some advantages of polynucleotide phosphorylase from E. coli in comparison with the M. luteus enzyme are discussed.
已开发出一种从大肠杆菌中分离多核苷酸磷酸化酶的新方法,包括离子交换色谱法和凝胶过滤法。该方法可使该酶纯化300倍,该酶无核酸酶和磷酸酶活性,可进一步用于寡核苷酸合成。结果表明,该酶在储存时会失去不依赖引物的活性,在没有氯化钠的情况下可用于进一步合成。添加氯化钠可刺激寡核苷酸链的延伸。文中讨论了大肠杆菌多核苷酸磷酸化酶相对于藤黄微球菌酶的一些优点。