Research Group Bioorganic Chemistry, Leibniz Institute for Plant Biochemistry, Weinberg 3, 06120, Halle (Saale), Germany.
Present address: Molecular Design and Engineering, Bayer AG, Aprather Weg 18 A, 42113, Wuppertal, Germany.
Angew Chem Int Ed Engl. 2023 Oct 9;62(41):e202307897. doi: 10.1002/anie.202307897. Epub 2023 Sep 4.
Fungal unspecific peroxygenases (UPOs) have gained substantial attention for their versatile oxyfunctionalization chemistry paired with impressive catalytic capabilities. A major drawback, however, remains their sensitivity towards their co-substrate hydrogen peroxide, necessitating the use of smart in situ hydrogen peroxide generation methods to enable efficient catalysis setups. Herein, we introduce flavin-containing protein photosensitizers as a new general tool for light-controlled in situ hydrogen peroxide production. By genetically fusing flavin binding fluorescent proteins and UPOs, we have created two virtually self-sufficient photo-enzymes (PhotUPO). Subsequent testing of a versatile substrate panel with the two divergent PhotUPOs revealed two stereoselective conversions. The catalytic performance of the fusion protein was optimized through enzyme and substrate loading variation, enabling up to 24300 turnover numbers (TONs) for the sulfoxidation of methyl phenyl sulfide. The PhotUPO concept was upscaled to a 100 mg substrate preparative scale, enabling the extraction of enantiomerically pure alcohol products.
真菌非特异性过氧化物酶（UPO）因其多功能的氧化官能化化学性质和令人印象深刻的催化能力而受到广泛关注。然而，其主要缺点仍然是对共底物过氧化氢的敏感性，这需要使用智能原位过氧化氢生成方法来实现有效的催化设置。在这里，我们引入黄素蛋白光敏剂作为一种新的通用工具，用于光控原位过氧化氢的产生。通过基因融合黄素结合荧光蛋白和 UPO，我们创建了两种几乎自给自足的光酶（PhotUPO）。随后用两种不同的 PhotUPO 对多功能底物进行测试，揭示了两种立体选择性转化。通过酶和底物加载变化对融合蛋白的催化性能进行了优化，使甲基苯基亚砜的氧化脱硫达到了 24300 的转化数（TON）。PhotUPO 概念被扩展到 100mg 底物的制备规模，能够提取对映体纯的醇产物。
