University of Chinese Academy of Sciences, Beijing 100049, China.
In Vitro Synthetic Biology Center, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China.
ACS Synth Biol. 2024 Aug 16;13(8):2611-2620. doi: 10.1021/acssynbio.4c00447. Epub 2024 Aug 2.
3-Hydroxypropionic acid (3-HP) is a highly sought-after platform chemical serving as a precursor to a variety of high value-added chemical products. In this study, we designed and constructed a novel light-powered synthetic enzymatic biosystem comprising acetyl-CoA ligase, acetyl-CoA carboxylase, malonyl-CoA reductase, and phosphotransferase to efficiently produce 3-HP through CO fixation from acetate, a cost-effective and readily available substrate. The system employed natural thylakoid membranes (TMs) for the regeneration of adenosine triphosphate and nicotinamide adenine dinucleotide phosphate. Comprehensive investigations were conducted on the effects of buffer solutions, substrate concentrations, enzyme loading levels, and TMs loading levels to optimize the yield of 3-HP. Following optimization, a production of 0.46 mM 3-HP was achieved within 6 h from an initial 0.5 mM acetate, with a yield nearing 92%. This work underscores the simplicity of 3-HP production via an biomanufacturing platform and highlights the potential for incorporating TMs as a sustainable and environmentally friendly approach in biomanufacturing processes.
3-羟基丙酸（3-HP）是一种备受追捧的平台化学品，可用作多种高附加值化学品的前体。在本研究中，我们设计并构建了一种新型的光驱动合成酶生物系统，该系统由乙酰辅酶 A 连接酶、乙酰辅酶 A 羧化酶、丙二酰辅酶 A 还原酶和磷酸转移酶组成，可通过固定 CO2 从乙酸（一种成本效益高且易于获得的底物）高效生产 3-HP。该系统利用天然类囊体膜（TMs）来再生三磷酸腺苷和烟酰胺腺嘌呤二核苷酸磷酸。我们全面研究了缓冲溶液、底物浓度、酶加载量和 TMs 加载量对 3-HP 产量的影响，以优化 3-HP 的产量。优化后，从初始 0.5 mM 的乙酸中，在 6 小时内可生产 0.46 mM 的 3-HP，产率接近 92%。这项工作突出了通过生物制造平台生产 3-HP 的简单性，并强调了将 TMs 作为生物制造过程中可持续和环保方法的潜力。
