Cis, -muconic acid (MA) is widely used as a key starting material in the synthesis of diverse polymers. The growing demand in these industries has led to an increased need for MA. Here, we constructed recombinant by systems metabolic engineering, which exhibit high efficiency in the production of MA. Firstly, the three major degradation pathways were disrupted in the MA production process. Subsequently, metabolic optimization strategies were predicted by computational design and the shikimate pathway was reconstructed, significantly enhancing its metabolic flux. Finally, through optimization and integration of key genes involved in MA production, the recombinant strain produced 88.2 g/L of MA with the yield of 0.30 mol/mol glucose in the 5 L bioreactor. This titer represents the highest reported titer achieved using glucose as the carbon source in current studies, and the yield is the highest reported for MA production from glucose in . Furthermore, to enable the utilization of more cost-effective glucose derived from corn straw hydrolysate, we subjected the strain to adaptive laboratory evolution in corn straw hydrolysate. Ultimately, we successfully achieved MA production in a high solid loading of corn straw hydrolysate (with the glucose concentration of 83.56 g/L), resulting in a titer of 19.9 g/L for MA, which is 4.1 times higher than that of the original strain. Additionally, the glucose yield was improved to 0.33 mol/mol. These provide possibilities for a greener and more sustainable production of MA.

中文翻译：

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用于高效生产顺式、顺式粘康酸的谷氨酸棒杆菌细胞工厂设计


顺式粘康酸 (MA) 广泛用作合成多种聚合物的关键起始原料。这些行业不断增长的需求导致对 MA 的需求增加。在这里，我们通过系统代谢工程构建了重组体，其在 MA 的生产中表现出高效率。首先，MA生产过程中三大降解途径被破坏。随后，通过计算设计预测代谢优化策略并重建莽草酸途径，显着增强其代谢通量。最后，通过对MA生产关键基因的优化和整合，重组菌株在5 L生物反应器中生产了88.2 g/L的MA，产量为0.30 mol/mol葡萄糖。该滴度代表了当前研究中使用葡萄糖作为碳源所达到的最高报道滴度，并且产率是 中报道的从葡萄糖生产 MA 的最高产量。此外，为了能够利用从玉米秸秆水解产物中提取的更具成本效益的葡萄糖，我们对该菌株在玉米秸秆水解产物中进行了适应性实验室进化。最终，我们在玉米秸秆水解液（葡萄糖浓度为83.56 g/L）的高固载量下成功实现了MA的生产，MA的效价达到19.9 g/L，比原始值提高了4.1倍拉紧。此外，葡萄糖产率提高至0.33 mol/mol。这些为更绿色、更可持续的 MA 生产提供了可能性。