Park Yu-Jin, Kim Dong-Myung
Department of Chemical Engineering and Applied Chemistry, Chungnam National University, Daejeon, South Korea.
Front Bioeng Biotechnol. 2021 Oct 27;9:778496. doi: 10.3389/fbioe.2021.778496. eCollection 2021.
One of the main advantages of a cell-free synthesis system is that the synthetic machinery of cells can be modularized and re-assembled for desired purposes. In this study, we attempted to combine the translational activity of extract with a heme synthesis pathway for the functional production of horseradish peroxidase (HRP). We first optimized the reaction conditions and the sequence of template DNA to enhance protein expression and folding. The reaction mixture was then supplemented with 5-aminolevulinic acid synthase to facilitate co-synthesis of the heme prosthetic group from glucose. Combining the different synthetic modules required for protein synthesis and cofactor generation led to successful production of functional HRP in a cell-free synthesis system.
无细胞合成系统的主要优点之一是细胞的合成机制可以模块化并重新组装以用于特定目的。在本研究中，我们试图将提取物的翻译活性与血红素合成途径相结合，以功能性生产辣根过氧化物酶（HRP）。我们首先优化了反应条件和模板DNA序列，以增强蛋白质表达和折叠。然后向反应混合物中添加5-氨基乙酰丙酸合酶，以促进从葡萄糖共合成血红素辅基。将蛋白质合成和辅因子生成所需的不同合成模块相结合，成功地在无细胞合成系统中生产出了功能性HRP。
