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The genome context of the ect genes was explored to identify proteins that are functionally associated with thesynthesisof ectoines;thespecialized aspartokinaseAsk_Ect and theregulatory protein EctR. This comprehensive in silico analysis was coupled with the biochemical characterization of ectoine hydroxylases from microorganisms that can colonize habitats with extremes in salinity (Halomonas elongata), pH (Alkalilimnicola ehrlichii,Acidiphilium cryptum), or temperature (Sphingopyxis alaskensis, Paenibacillus lautus) or that produce hydroxyectoine very efficiently over ectoine (Pseudomonasstutzeri).These six ectoine hydroxylasesall possesssimilar kinetic parametersfor their substrates but exhibit different temperature stabilities and differ in their tolerance to salts. We also report the crystal structure of the Virgibacillus salexigens EctD protein in its apo-form, thereby revealing that the iron-free structure exists already in a pre-set configuration to incorporate the iron catalyst. Collectively, our work definesthe taxonomic distribution and salient biochemical properties of the ectoine hydroxylase protein family and contributes to the understanding of its structure. |
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| 为了找出与合成四氢嘧啶功能相关的蛋白质，作者对表基因的基因组前后，特别是天冬氨酸激酶Ask_Ect及其调节蛋白 EctR进行了研究。综合的计算机模拟分析与四氢嘧啶羟化酶的生化特性相结合。这些酶来自于能定植于极端盐（长卤単胞菌）、pH(埃里希氏碱湖沼菌、隐藏嗜酸菌）、或温度（阿拉斯加鞘氨醇単胞菌，灿烂芽孢样杆菌）生境，或能很有效地利用四氢嘧啶产生羟化四氢嘧啶（施氏假单胞菌）的微生物。这六种四氢嘧啶羟化酶对其底物都有相似的动力学参数，它们对温度的稳定性和对盐的耐受性不同。我们也报告需盐枝芽孢杆菌EctD 脱辅基蛋白的晶体结构，进而说明无铁结构已存在于引入铁离子催化剂之前的结构中。总之，我们的研究确定了四氢嘧啶羟化酶在分类学上的分布和突出的生化特性，并对其结构的理解做出了贡献。 |
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