BackgroundThe viability of biorefinery is readily acknowledged by integration of the protein-based platform into the existing production process that manages biomass. Biomass-dependent processes generate large quantities of proteins in waste streams which are commonly rich in glutamate (Glu) and aspartate (Asp). MethodsThe protein biorefinery platform was developed for production of succinic acid (SA). This was carried out by deciphering the Glu and Asp metabolism for targeted manipulation in Escherichia coli. It was found that citrate synthase and NADP-dependent malic enzyme involved in the tricarboxylic acid cycle played a positive role in the synthesis of SA. By adjusting the activity of two enzymes, the producer strain enabled production of SA reaching 76.3% of the theoretical yield based on 10 g/L Glu and 10 g/L Asp. Furthermore, the SA production accounting for 72.7% of the theoretical yield could be obtained for the strain grown on soybean meals hydrolysate. Significant findingsIn this study, key reaction steps were identified in Glu and Asp metabolism. It provides informative inputs for the design of synthetic routes that convert Glu and Asp to value-added products of interest, which opens up a new avenue of biorefinery based on proteins.