FeFe]-氢化酶的氧敏感性:酶内与酶外活性位点模拟物的比较研究

IF 2.9 3区 化学 Q3 CHEMISTRY, PHYSICAL
Shanika Yadav, Rieke Haas, Esma Birsen Boydas, Michael Roemelt, Thomas Happe, Ulf-Peter Apfel and Sven T. Stripp
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引用次数: 0

摘要

[FeFe]-氢化酶是自然界最有效的质子还原和氢氧化酶。然而,这种金属酶对氧气的敏感性阻碍了生物技术的应用,而且有氧失活的机理也不甚明了。在此,我们通过红外光谱、莫斯鲍尔光谱和核磁共振光谱,探讨了[FeFe]-氢化酶有机金属活性位点辅助因子的四种模拟物[Fe2(apt)(CO)6-x(CN)x]x-和[Fe2(pdt)(CO)6-x(CN)x]x-(x = 1,2)以及该酶相应辅助因子变体的氧敏感性。此外,我们还描述了活性位点前体复合物 Fe2(adt)(CO)6 的直接合成配方。我们的数据表明,作为天然活性位点辅助因子合成前体的氨基二硫酸盐(adt)复合物对氧最为敏感。这一观察结果凸显了质子转移在有氧失活过程中的重要作用,并在 DFT 计算的支持下,有助于确定负责的活性氧物种(ROS)。此外,我们还发现,铁离子的配体环境对与 O2 和 ROS(如超氧化物和 H2O2)的反应性有着至关重要的影响,因为随着配体从 CO 交换到 CN-,对氧的敏感性也随之增加。在模型复合物中观察到的有氧失活趋势与各自的酶变体一致。根据实验和计算数据,建立了[FeFe]-氢化酶与 O2 初始反应的模型。我们的研究强调了模型系统在理解生物催化过程中的相关性,并验证了模型系统作为阐明氧气诱导的[FeFe]-氢化酶失活化学反应的重要工具的潜力。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Oxygen sensitivity of [FeFe]-hydrogenase: a comparative study of active site mimics inside vs. outside the enzyme†

Oxygen sensitivity of [FeFe]-hydrogenase: a comparative study of active site mimics inside vs. outside the enzyme†

[FeFe]-hydrogenase is nature's most efficient proton reducing and H2-oxidizing enzyme. However, biotechnological applications are hampered by the O2 sensitivity of this metalloenzyme, and the mechanism of aerobic deactivation is not well understood. Here, we explore the oxygen sensitivity of four mimics of the organometallic active site cofactor of [FeFe]-hydrogenase, [Fe2(adt)(CO)6−x(CN)x]x and [Fe2(pdt)(CO)6−x(CN)x]x (x = 1, 2) as well as the corresponding cofactor variants of the enzyme by means of infrared, Mössbauer, and NMR spectroscopy. Additionally, we describe a straightforward synthetic recipe for the active site precursor complex Fe2(adt)(CO)6. Our data indicate that the aminodithiolate (adt) complex, which is the synthetic precursor of the natural active site cofactor, is most oxygen sensitive. This observation highlights the significance of proton transfer in aerobic deactivation, and supported by DFT calculations facilitates an identification of the responsible reactive oxygen species (ROS). Moreover, we show that the ligand environment of the iron ions critically influences the reactivity with O2 and ROS like superoxide and H2O2 as the oxygen sensitivity increases with the exchange of ligands from CO to CN. The trends in aerobic deactivation observed for the model complexes are in line with the respective enzyme variants. Based on experimental and computational data, a model for the initial reaction of [FeFe]-hydrogenase with O2 is developed. Our study underscores the relevance of model systems in understanding biocatalysis and validates their potential as important tools for elucidating the chemistry of oxygen-induced deactivation of [FeFe]-hydrogenase.

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来源期刊
Physical Chemistry Chemical Physics
Physical Chemistry Chemical Physics 化学-物理:原子、分子和化学物理
CiteScore
5.50
自引率
9.10%
发文量
2675
审稿时长
2.0 months
期刊介绍: Physical Chemistry Chemical Physics (PCCP) is an international journal co-owned by 19 physical chemistry and physics societies from around the world. This journal publishes original, cutting-edge research in physical chemistry, chemical physics and biophysical chemistry. To be suitable for publication in PCCP, articles must include significant innovation and/or insight into physical chemistry; this is the most important criterion that reviewers and Editors will judge against when evaluating submissions. The journal has a broad scope and welcomes contributions spanning experiment, theory, computation and data science. Topical coverage includes spectroscopy, dynamics, kinetics, statistical mechanics, thermodynamics, electrochemistry, catalysis, surface science, quantum mechanics, quantum computing and machine learning. Interdisciplinary research areas such as polymers and soft matter, materials, nanoscience, energy, surfaces/interfaces, and biophysical chemistry are welcomed if they demonstrate significant innovation and/or insight into physical chemistry. Joined experimental/theoretical studies are particularly appreciated when complementary and based on up-to-date approaches.
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