Artificial Light-Harvesting Complexes Enable Rieske Oxygenase Catalyzed Hydroxylations in Non-Photosynthetic cells
Artificial Light-Harvesting Complexes Enable Rieske Oxygenase Catalyzed Hydroxylations in Non-Photosynthetic cells
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DOI：
10.1002/anie.201914519
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发表时间：
2020-01-24
影响因子：
16.6
通讯作者：
Schmidt, Sandy
中科院分区：
文献类型：
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作者：
Oezgen, F. Feyza;Runda, Michael E.;Schmidt, Sandy
在这项研究中，我们耦合了一个完善的全细胞系统的基础上，E。大肠杆菌通过捕光复合物对Rieske加氧酶（RO）催化的体内羟基化。虽然这些酶代表非常有前途的生物催化剂，但它们的实用性受到它们对NAD（P）H的依赖性以及它们在无细胞系统中的多组分性质和内在不稳定性的阻碍。为了探索E.大肠杆菌作为人工光合作用的底盘，并且由于报道的RO的不稳定性，我们使用这些具有挑战性的酶作为模型系统。光驱动的方法依赖于光捕获复合物，如曙红Y，5（6）-羧基曙红，和玫瑰红和牺牲电子供体（EDTA，MOPS和MES），很容易被细胞吸收。得到的产物形成高达1.3 g L-1和高达1.6 mm h（-1）的速率表明，这是一个典型的全细胞转化在E。杆菌这种光催化合成的适用性已经得到证明，并代表了光诱导RO系统的第一个例子。
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.