Tang Shuai, Sun Li-Juan, Pan Ai-Qun, Huang Jun, Wang Huamin, Lin Ying-Wu
School of Chemistry and Chemical Engineering, University of South China, Hengyang 421001, China.
Hengyang Medical College, University of South China, Hengyang 421001, China.
Org Biomol Chem. 2023 Dec 13;21(48):9603-9609. doi: 10.1039/d3ob01687k.
Significant efforts have been made in the design of artificial metalloenzymes. Myoglobin (Mb), an O carrier, has been engineered to exhibit different functions. Herein, we applied a series of engineered Mb mutants with peroxidase activity for biosynthesis of clofazimine (CFZ), a potential drug with a broad-spectrum antiviral activity, by integration with chemical synthesis. Two of those mutants, F43Y Mb and F43Y/T67R Mb, have been shown to efficiently catalyze the oxidative coupling of 2--(4-chlorophenyl) benzene-1,2-diamine (-4-CPBDA) in the presence of HO, with 97% yields. The overall catalytic efficiency (/) is 46-fold and 82-fold higher than that of WT Mb, respectively. By further combination of this reaction with chemical synthesis, the production of CFZ was accomplished with an isolated yield of 72%. These results showed that engineered Mbs containing the Tyr-heme cross-link (F43Y Mb and F43Y/T67R Mb) exhibit enhanced activity in the oxidative coupling reaction. This study also indicates that the combination of biocatalysis and chemical synthesis avoids the need for the separation of intermediate products, which offers a convenient approach for the total synthesis of the biological compound CFZ.
在人工金属酶的设计方面已经做出了巨大努力。肌红蛋白（Mb）作为一种氧载体，已被改造以展现不同功能。在此，我们通过与化学合成相结合，应用了一系列具有过氧化物酶活性的工程化Mb突变体来生物合成氯法齐明（CFZ），一种具有广谱抗病毒活性的潜在药物。其中两个突变体，F43Y Mb和F43Y/T67R Mb，已被证明在过氧化氢存在下能有效催化2 - （4 - 氯苯基）苯 - 1,2 - 二胺（-4 - CPBDA）的氧化偶联反应，产率达97%。总体催化效率（/）分别比野生型Mb高46倍和82倍。通过将该反应与化学合成进一步结合，实现了CFZ的生产，分离产率为72%。这些结果表明，含有酪氨酸 - 血红素交联结构的工程化Mb（F43Y Mb和F43Y/T67R Mb）在氧化偶联反应中表现出增强的活性。这项研究还表明，生物催化与化学合成的结合避免了中间产物分离的需要，为生物化合物CFZ的全合成提供了一种便捷方法。
