The cbiS gene of the archaeon Methanopyrus kandleri AV19 encodes a bifunctional enzyme with adenosylcobinamide amidohydrolase and alpha-ribazole-phosphate phosphatase activities.
The cbiS gene of the archaeon Methanopyrus kandleri AV19 encodes a bifunctional enzyme with adenosylcobinamide amidohydrolase and alpha-ribazole-phosphate phosphatase activities.
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古细菌 Methanopyrus kandleri AV19 的 cbiS 基因编码一种具有腺苷酰胺酰胺水解酶和 α-利唑-磷酸磷酸酶活性的双功能酶。
DOI：
10.1128/jb.00227-06
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发表时间：
2006
影响因子：
3.2
通讯作者：
Escalante-Semerena,JorgeC
中科院分区：
文献类型：
--
作者：
Woodson,JesseD;Escalante-Semerena,JorgeC
在这里，我们报告了来自超嗜热产甲烷古菌Methanopyrus kandleriAV 19的双功能α-Ribazole-P（α-RP）磷酸酶、腺苷二氨酰胺（AdoCbi）酰胺水解酶CbiS酶的初步生化特征。该cbiS基因编码具有两个不同片段的39-kDa蛋白质，其中一个与CbiZ，EC 3.5.1.90）酶同源，另一个与最近发现的古细菌α-RP磷酸酶（CobZ，EC 3.1.3.73）酶同源。CbiS功能恢复了肠道沙门氏菌菌株中Cbi补救和α-RP磷酸酶活性，其中任一步骤都被阻断。当它们被分别克隆时，这两个半体的双基因保留了它们在体内的功能。CbiS酶在大肠杆菌中过量生产，并分离到>95%的均一性。高效液相色谱法、紫外可见光谱法和质谱法分别确定α-ribazole和cobyric acid为磷酸酶和酰胺水解酶反应的产物。对CbiZ和CobZ酶在某些古细菌中融合的原因进行了讨论。
Here we report the initial biochemical characterization of the bifunctional α-ribazole-P (α-RP) phosphatase, adenosylcobinamide (AdoCbi) amidohydrolase CbiS enzyme from the hyperthermophilic methanogenic archaeonMethanopyrus kandleriAV19. ThecbiSgene encodes a 39-kDa protein with two distinct segments, one of which is homologous to the AdoCbi amidohydrolase (CbiZ, EC 3.5.1.90) enzyme and the other of which is homologous to the recently discovered archaeal α-RP phosphatase (CobZ, EC 3.1.3.73) enzyme. CbiS function restored AdoCbi salvaging and α-RP phosphatase activity in strains of the bacteriumSalmonella entericawhere either step was blocked. The two halves of thecbiSgenes retained their function in vivo when they were cloned separately. The CbiS enzyme was overproduced inEscherichia coliand was isolated to >95% homogeneity. High-performance liquid chromatography, UV-visible spectroscopy, and mass spectroscopy established α-ribazole and cobyric acid as the products of the phosphatase and amidohydrolase reactions, respectively. Reasons why the CbiZ and CobZ enzymes are fused in some archaea are discussed.