Efficient production of α-ketoglutaric acid using an economical double-strain cultivation and catalysis system

The whole-cell catalysis strategy of alpha-ketoglutaric acid (α-KG) production from L-glutamic acid (L-Glu) using recombinant Escherichia coli, in which L-glutamate oxidase (LGox) was over-expressed, has replaced the traditional chemical synthesis strategy. However, large amounts of toxic by-product, H₂O₂, should be eliminated through co-expressing catalase (Cat), thus severely increasing burden in cells. To efficiently and economically produce α-KG, here, the genes SpLGox (from Streptomyces platensis NTU3304) and SlCat (from Streptomyces lividans TK24) were inserted into the low-dosage-IPTG (Isopropyl β-D-Thiogalactoside) inducible expression system, constructed in our previous work, in E. coli, respectively. Besides, a double-strain catalysis system was established and optimized to produce α-KG, and the productivity of α-KG was increased 97% compared with that through single strain catalysis. Finally, a double-strain cultivation strategy was designed and employed to simplify the scale-up fermentation. Using the optimized whole-cell biocatalyst conditions (pH 7.0, 35 °C), majority of the L-glutamic acid was transformed into α-KG and the titer reached 95.4 g/L after 6 h with the highest productivity at present. Therefore, this strategy may efficiently and cost-effectively produce α-KG, enhancing its potential for industrial applications. KEY POINTS: • SpLGox and SlCat were over-expressed to catalyze L-Glu to α-KG and eliminate by-product H₂O₂, respectively. • Double-strain cultivation and catalysis system can efficiently and cost-effectively produce α-KG from L-Glu.