Enhanced production of maltobionic acid by a metabolically engineered Escherichia coli incapable of maltose utilization.

Abstract

To produce maltobionic acid (MBA) from maltose in Escherichia coli, we recombinantly expressed a glucose dehydrogenase gene (gdh1) from Enterobacter cloacae and a pyrroloquinoline quinone (PQQ) synthesis gene cluster (pqqFABCDEMIH) from Pseudomonas taetrolens. Although the recombinant E. coli strain (E. coli [pKK-ECGDH1 + pACYC-PQQ]) successfully produced MBA from maltose, the yield of MBA was rather low, indicating that E. coli has other maltose utilization pathways. Amylomaltase (MalQ) is the first enzyme in the maltose utilization pathway in E. coli. To investigate the potential role of MalQ on MBA production, E. coli malQ was inactivated. The culturing of the recombinant E. coli strain (E. coli ∆malQ [pKK-ECGDH1 + pACYC-PQQ]) in a flask resulted in higher MBA production titer, yield, and productivity (209.3g/L, 100%, and 1.1g/L/h, respectively) than those of E. coli [pKK-ECGDH1 + pACYC-PQQ] (162.1g/L, 77.4%, and 0.5g/L/h, respectively), indicating that the MalQ inactivation was highly effective in improving the MBA production ability of E. coli. After fermentation using 5-L bioreactor, MBA production titer, yield, and productivity of the recombinant E. coli strain were 209.3g/L, 100%, and 1.5g/L/h, respectively, which were 1.3-, 1.3-, 2.3-fold higher than those of E. coli [pKK-ECGDH1 + pACYC-PQQ] (167.3g/L, 79.9%, and 0.65g/L/h), respectively. Thus, our results provide an important foundation for efficient MBA production using recombinant E. coli strain.