Abstract

Thymidine phosphorylase (deoA) and purine nucleoside phosphorylase (deoD) genes from the extremophilic bacterium Halomonas chromatireducens AGD 8-3 have been cloned. Expression plasmids were constructed, and highly efficient recombinant producer strains were obtained for these proteins. Recombinant nucleoside phosphorylases were isolated by ion-exchange chromatography in a homogeneous state, and their physical and enzymatic properties were studied. It was shown that the studied thymidine phosphorylase (HrTPP) and purine nucleoside phosphorylase (HrPNP) form the dimeric and hexameric forms, respectively. It was shown that the specific activity of HrTPP from the extremophilic bacterium H. chromatireducens AGD 8-3 was higher relative to thymidine (in comparison with its counterpart from E. coli).

中文翻译：

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嗜热嗜色菌嗜盐嗜铬菌AGD 8-3的核苷磷酸化酶基因的克隆及其重组酶的制备及酶学性质的研究

摘要

已经克隆了嗜极端嗜热菌嗜铬嗜铬菌AGD 8-3的胸苷磷酸化酶（deoA）和嘌呤核苷磷酸化酶（deoD）基因。构建表达质粒，并获得这些蛋白的高效重组生产菌株。通过离子交换色谱以均相状态分离重组核苷磷酸化酶，并研究其物理和酶学性质。结果表明，所研究的胸苷磷酸化酶（HrTPP）和嘌呤核苷磷酸化酶（HrPNP）分别形成二聚体和六聚体形式。结果表明，从细菌嗜极HrTPP的比活性H. chromatireducensAGD 8-3相对于胸腺嘧啶核苷更高（与其来自大肠杆菌的对应物相比）。