Because only 0.01% of prokaryotic genospecies can be cultured and in situ observations are often impracticable, culture-independent methods are required to understand microbial life and harness potential applications of microbes. Here, we report a methodology for the production of proteins with desired functions based on single amplified genomes (SAGs) from unculturable species. We use this method to resurrect an alcohol dehydrogenase (ADH/D1) from an uncharacterized halo-thermophilic archaeon collected from a brine pool at the bottom of the Red Sea. Our crystal structure of 5,6-dihydroxy NADPH-bound ADH/D1 combined with biochemical analyses reveal the molecular features of its halo-thermophily, its unique habitat adaptations, and its possible reaction mechanism for atypical oxygen activation. Our strategy offers a general guide for using SAGs as a source for scientific and industrial investigations of “microbial dark matter.”

中文翻译：

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从单个扩增基因组的极端嗜盐池池酒精脱氢酶的鉴定和实验表征

因为只能原位培养0.01％的原核基因种观察通常是不可行的，需要采用与培养无关的方法来了解微生物的生命并利用微生物的潜在应用。在这里，我们报告了一种基于无法培养的物种的单个扩增基因组（SAG）产生具有所需功能的蛋白质的方法。我们使用这种方法从红海底部盐水池中收集到的未表征的卤代嗜热古细菌中复活醇脱氢酶（ADH / D1）。我们的5,6-二羟基NADPH结合的ADH ​​/ D1的晶体结构与生化分析相结合，揭示了其光晕嗜热的分子特征，其独特的生境适应性以及其非典型氧活化的可能反应机理。