The O2-stable [FeFe]-hydrogenase CbA5H reveals high resilience against organic solvents†

IF 4.4 3区 化学 Q2 CHEMISTRY, PHYSICAL
Martin Gerbaulet , Anja Hemschemeier , Thomas Happe
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Abstract

[FeFe]-Hydrogenases are highly efficient hydrogen-(H2) converting enzymes which play pivotal roles for H2 cycling in natural habitats, but which are also of interest for sustainable approaches to generate or employ H2 gas. [FeFe]-Hydrogenases harbor a unique active site metal cofactor, the H-cluster, whose di-iron site by itself is nearly inactive but, as part of the protein, allows high turnover rates. Understanding this essential interplay of protein and co-factor might help to install [FeFe]-hydrogenases in biotechnological applications. The catalytic unit of the H-Cluster can be synthesized chemically and incorporated into [FeFe]-hydrogenase precursors, which allows to introduce non-natural metals or ligands and study their impact on catalytic activity. However, these compounds are often not water-soluble and have to be added to the proteins in solvents known to destabilize polypeptides. The resilience of [FeFe]-hydrogenases against organic solvents has hardly been investigated. To address this knowledge gap, we characterized the stability of the [FeFe]-hydrogenase CbA5H from Clostridium beijerinckii in several organic solvents (dimethylsulfoxide (DMSO), acetone, acetonitrile as well as water-miscible short-chain alcohols). These solvents are required to dissolve co-factor analogues and are also employed in chemical syntheses that might be combined with biocatalysts such as hydrogenases for more sustainable industrial processes. In the medium time-frame, CbA5H is remarkably stable in high concentrations of acetone and acetonitrile and also withstands intermediate concentrations of DMSO, ethanol and methanol. Combined with the unusual O2 stability and high temperature and pressure tolerance, this makes CbA5H a candidate for its use in non-aqueous reaction environments.

Abstract Image

o2稳定的[FeFe]-氢化酶CbA5H显示出对有机溶剂的高弹性†
[FeFe]-氢化酶是一种高效的氢(H2)转化酶,在自然栖息地的H2循环中起着关键作用,但它也对产生或利用H2气体的可持续方法感兴趣。[FeFe]-氢化酶具有独特的活性位点金属辅助因子h簇,其双铁位点本身几乎不活跃,但作为蛋白质的一部分,允许高周转率。了解蛋白质和辅助因子的这种基本相互作用可能有助于在生物技术应用中安装[FeFe]-氢化酶。h -簇的催化单元可以化学合成并掺入[FeFe]-氢化酶前体中,这允许引入非天然金属或配体并研究它们对催化活性的影响。然而,这些化合物通常不是水溶性的,必须添加到已知会破坏多肽稳定的溶剂中的蛋白质中。[FeFe]-氢化酶对有机溶剂的弹性研究很少。为了解决这一知识空白,我们表征了贝氏梭菌[FeFe]-氢化酶CbA5H在几种有机溶剂(二甲基亚砜(DMSO)、丙酮、乙腈以及与水混溶的短链醇)中的稳定性。这些溶剂用于溶解辅助因子类似物,也用于化学合成,可能与生物催化剂(如氢化酶)结合,以实现更可持续的工业过程。在中等时间范围内,CbA5H在高浓度丙酮和乙腈中表现出显著的稳定性,也能承受中等浓度的DMSO、乙醇和甲醇。结合不同寻常的氧稳定性和高温耐压性,这使得CbA5H成为非水反应环境中使用的候选者。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Catalysis Science & Technology
Catalysis Science & Technology CHEMISTRY, PHYSICAL-
CiteScore
8.70
自引率
6.00%
发文量
587
审稿时长
1.5 months
期刊介绍: A multidisciplinary journal focusing on cutting edge research across all fundamental science and technological aspects of catalysis. Editor-in-chief: Bert Weckhuysen Impact factor: 5.0 Time to first decision (peer reviewed only): 31 days
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