来自莱氏乳杆菌的腺苷钴胺素依赖性核糖核苷酸还原酶的克隆、测序及表达
Cloning, sequencing, and expression of the adenosylcobalamin-dependent ribonucleotide reductase from Lactobacillus leichmannii.
作者信息
Booker S, Stubbe J
机构信息
Department of Chemistry, Massachusetts Institute of Technology, Cambridge 02139.
出版信息
Proc Natl Acad Sci U S A. 1993 Sep 15;90(18):8352-6. doi: 10.1073/pnas.90.18.8352.
Abstract
Ribonucleoside-triphosphate reductase (RTPR, EC 1.17.4.2) from Lactobacillus leichmannii, a monomeric adenosylcobalamin-requiring enzyme, catalyzes the conversion of nucleoside triphosphates to deoxynucleoside triphosphates. The gene for this enzyme has been cloned and sequenced. In contrast to expectations based on mechanistic considerations, there is no statistically significant sequence homology with the Escherichia coli reductase that requires a dinuclear-iron center and tyrosyl radical cofactor. The RTPR has been overexpressed and purified to homogeneity, yielding 90 mg of protein from 2.5 g of bacteria. Initial characterization of the recombinant RTPR indicates that its properties are identical to those of the RTPR isolated from L. leichmannii.
摘要
来自赖氏乳杆菌的核糖核苷三磷酸还原酶（RTPR，EC 1.17.4.2）是一种需要腺苷钴胺素的单体酶，催化核苷三磷酸转化为脱氧核苷三磷酸。该酶的基因已被克隆和测序。与基于机理考虑的预期相反，它与需要双核铁中心和酪氨酰自由基辅因子的大肠杆菌还原酶没有统计学上显著的序列同源性。RTPR已被过量表达并纯化至同质，从2.5克细菌中获得了90毫克蛋白质。重组RTPR的初步表征表明，其性质与从赖氏乳杆菌中分离的RTPR相同。
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