Elling L, Kula M R
Institut für Enzymtechnologie, Heinrich-Heine-Universität Düsseldorf im Forschungszentrum Jülich, Germany.
J Biotechnol. 1994 May 15;34(2):157-63. doi: 10.1016/0168-1656(94)90085-x.
UDP-glucose pyrophosphorylase was purified from germinated barley (malt) using anion exchange and hydrophobic interaction chromatography followed by preparative gel filtration. Gel filtration and SDS-PAGE revealed a molecular mass of 51 to 54 kDa for the monomeric protein. Microsequencing of the blotted protein by Edman degradation gave 20 N-terminal amino acids. UDP-glucose pyrophosphorylase from malt could be markedly stabilized by the addition of bovine serum albumin. The enzyme preparation is free of contaminating nucleoside triphosphatases (UTPases) and can be utilized for the enzymatic synthesis of activated sugars.
利用阴离子交换和疏水相互作用色谱法，随后进行制备性凝胶过滤，从发芽大麦（麦芽）中纯化出尿苷二磷酸葡萄糖焦磷酸化酶。凝胶过滤和十二烷基硫酸钠-聚丙烯酰胺凝胶电泳显示该单体蛋白的分子量为51至54千道尔顿。通过埃德曼降解法对印迹蛋白进行微量测序得到了20个N端氨基酸。添加牛血清白蛋白可显著稳定麦芽中的尿苷二磷酸葡萄糖焦磷酸化酶。该酶制剂不含污染性核苷三磷酸酶（UTP酶），可用于活化糖的酶促合成。
