Corynebacterium glutamicum cell factory design for the efficient production of cis, cis-muconic acid

• Published in: Metabolic engineering 2024-03, Vol.82, p.225-237
• Main Authors: Li, Menglei, Chen, Jiayao, He, Keqin, Su, Changsheng, Wu, Yilu, Tan, Tianwei
• Format: Article
• Language: English
• Subjects: ALE; Corynebacterium glutamicum; Muconic acid; Shikimate pathway
• ISSN: 1096-7176; 1096-7184
• DOI: 10.1016/j.ymben.2024.02.005

Cis, 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 Corynebacterium glutamicum 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 Corynebacterium glutamicum. 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. •Cis, cis-muconic acid is a key platform substance in industrial production for important chemicals.•Metabolic simulation predicts effective gene manipulation targets.•Enhancing metabolic flux through the reconstruction of the shikimate pathway.•The highest cis, cis-muconic acid titer (88.2 g/L) was achieved in C. glutamicum using glucose as the carbon source.•Adaptive laboratory evolution enhances the fermentation capability of the strain in corn straw hydrolysate.