Rubredoxin Protein Scaffolds Sourced from Diverse Environmental Niches as an Artificial Hydrogenase Platform
Rubredoxin Protein Scaffolds Sourced from Diverse Environmental Niches as an Artificial Hydrogenase Platform
复制标题
红氧还蛋白蛋白支架源自不同的环境生态位，作为人工氢化酶平台
DOI：
10.1021/acs.biochem.3c00249
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发表时间：
2023
期刊：
影响因子：
2.9
通讯作者：
Shafaat, Hannah S.
中科院分区：
文献类型：
--
作者：
Wertz, Ashlee E.;Teptarakulkarn, Pathorn;Stein, Riley E.;Moore, Peter J.;Shafaat, Hannah S.
来自脱硫脱硫弧菌的镍取代红氧还蛋白（NiRd）在结构和功能上都是[NiFe]氢化酶的模拟物。然而，在营业额的频率和超电位的改善，需要竞争对手的本地[NiFe]氢化酶。表征的NiRd突变体库的变化，在二级协调领域的蛋白质动力学在调节活性中发挥了重要作用。在这项工作中，从不同的生物体中选择rubredoxin支架来研究远端序列变异对催化活性的影响。结果发现，尽管电化学催化活性在整个系列中仅受到轻微影响，但来自嗜冷生物的Rd序列表现出显著更高水平的溶液相氢产生。此外，Eyring分析表明，催化活化特性与母体生物的生长温度有关，这意味着在天然存在的酶中经常观察到的母体生物环境和催化活性之间的一般相关性也可以在人工酶中观察到。从栖息于不同环境，特别是低温环境的宿主中选择蛋白质支架，代表了工程化人工金属酶的另一种方法。
Nickel-substituted rubredoxin (NiRd) fromDesulfovibrio desulfuricanshas previously been shown to act as both a structural and functional mimic of the [NiFe] hydrogenase. However, improvements both in turnover frequency and overpotential are needed to rival the native [NiFe] hydrogenase enzymes. Characterization of a library of NiRd mutants with variations in the secondary coordination sphere suggested that protein dynamics played a substantial role in modulating activity. In this work, rubredoxin scaffolds were selected from diverse organisms to study the effects of distal sequence variation on catalytic activity. It was found that though electrochemical catalytic activity was only slightly impacted across the series, the Rd sequence from a psychrophilic organism exhibited substantially higher levels of solution-phase hydrogen production. Additionally, Eyring analyses suggest that catalytic activation properties relate to the growth temperature of the parent organism, implying that the general correlation between the parent organism environment and catalytic activity often seen in naturally occurring enzymes may also be observed in artificial enzymes. Selecting protein scaffolds from hosts that inhabit diverse environments, particularly low-temperature environments, represents an alternative approach for engineering artificial metalloenzymes.