Engineering of itaconic acid pathway via co-localization of CadA and AcnA in recombinant Escherichia coli

• Published in: Biotechnology letters 2024-08, Vol.46 (4), p.593-600
• Main Authors: Tran, Kim-Ngan T., Jeong, Jaehoon, Hong, Soon Ho
• Format: Article
• Language: English
• Subjects: Acids; Applied Microbiology; Biochemistry; Biomedical and Life Sciences; Biotechnology; Carbon; Cloning; E coli; Engineering; Enzymes; Escherichia coli; Escherichia coli - genetics; Escherichia coli - metabolism; Escherichia coli Proteins - genetics; Escherichia coli Proteins - metabolism; Fermentation; Genes; Glucose; Glycerol; Industrial applications; Itaconic acid; Life Sciences; Ligands; Localization; Metabolic Engineering - methods; Metabolism; Metabolites; Microbiology; Original Research Paper; Plasmids; Prepolymers; Productivity; Proteins; Scaffolds; Substrates; Succinates - metabolism
• ISSN: 0141-5492; 1573-6776; 1573-6776
• DOI: 10.1007/s10529-024-03496-x

Itaconic acid is an excellent polymeric precursor with a wide range of industrial applications. The efficient production of itaconate from various renewable substrates was demonstrated by engineered Escherichia coli . However, limitation in the itaconic acid precursor supply was revealed by finding out the key intermediate of the tricarboxylic acid in the itaconic acid pathway. Efforts of enhancing the cis-aconitate flux and preserving the isocitrate pool to increase itaconic acid productivity are required. In this study, we introduce a synthetic protein scaffold system between CadA and AcnA to physically combine the two enzymes. Through the introduction of a synthetic protein scaffold, 2.1 g L −1 of itaconic acid was produced at pH 7 and 37 °C. By fermentation, 20.1 g L −1 for 48 h of itaconic acid was produced with a yield of 0.34 g g −1 glycerol. These results suggest that carbon flux was successfully increased itaconic acid productivity.