In this study, we coupled a well-established whole-cell system based on E. coli via light-harvesting complexes to Rieske oxygenase (RO)-catalyzed hydroxylations in vivo. Although these enzymes represent very promising biocatalysts, their practical applicability is hampered by their dependency on NAD(P)H as well as their multicomponent nature and intrinsic instability in cell-free systems. In order to explore the boundaries of E. coli as chassis for artificial photosynthesis, and due to the reported instability of ROs, we used these challenging enzymes as a model system. The light-driven approach relies on light-harvesting complexes such as eosin Y, 5(6)-carboxyeosin, and rose bengal and sacrificial electron donors (EDTA, MOPS, and MES) that were easily taken up by the cells. The obtained product formations of up to 1.3 g L-1 and rates of up to 1.6 mm h-1 demonstrate that this is a comparable approach to typical whole-cell transformations in E. coli. The applicability of this photocatalytic synthesis has been demonstrated and represents the first example of a photoinduced RO system.

中文翻译：

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人工采光复合物可在非光合作用细胞中实现Rieske加氧酶催化的羟化反应。

在这项研究中，我们通过光收集复合物将基于大肠杆菌的成熟的全细胞系统与Rieske加氧酶（RO）催化的体内羟基化反应相结合。尽管这些酶代表了非常有前途的生物催化剂，但它们对NAD（P）H的依赖性以及它们的多组分性质和在无细胞系统中的固有不稳定性阻碍了它们的实际应用。为了探索作为人工光合作用底盘的大肠杆菌的边界，并且由于报道的RO不稳定，我们使用了这些具有挑战性的酶作为模型系统。光驱动方法依赖于光吸收复合物，如曙红Y，5（6）-羧基曙红，玫瑰红和牺牲性电子供体（EDTA，MOPS和MES），这些复合物很容易被细胞吸收。所获得的产物形成量高达1.3 g L-1，速率高达1。6 mm h-1证明这是大肠杆菌中典型的全细胞转化的可比方法。已经证明了这种光催化合成的适用性，并且代表了光致反渗透系统的第一个例子。