Production of itaconic acid from acetate by engineering acid‐tolerant Escherichia coli W

Utilization of abundant and cheap carbon sources can effectively reduce the production cost and enhance the economic feasibility. Acetate is a promising carbon source to achieve cost‐effective microbial processes. In this study, we engineered an Escherichia coli strain to produce itaconic acid from acetate. As acetate is known to inhibit cell growth, we initially screened for a strain with a high tolerance to 10 g/L of acetate in the medium, and the W strain was selected as the host. Subsequently, the WC strain was obtained by overexpression of cad (encoding cis‐aconitate decarboxylase) using a synthetic promoter and 5′ UTR. However, the WC strain produced only 0.13 g/L itaconic acid because of low acetate uptake. To improve the production, the acetate assimilating pathway and glyoxylate shunt pathway were amplified by overexpression of pathway genes as well as its deregulation. The resulting strain, WCIAG4 produced 3.57 g/L itaconic acid (16.1% of theoretical maximum yield) after 88 hr of fermentation with rapid acetate assimilation. These efforts support that acetate can be a potential feedstock for biochemical production with engineered E. coli. Production of itaconic acid from acetate can be promising strategy to reduce the production cost and enhance the economic feasibility. However, sole expression of synthetic pathway enzyme, cad in E. coli was inefficient in producing itaconic acid due to low acetate assimilation and insufficient glyoxylate shunt activity. Activation of acetate assimilating pathway and glyoxylate shunt could lead significantly increased acetate uptake and itaconic acid production, suggesting acetate can be a potential feedstock for biochemical production.