Combined C-assisted metabolomics and metabolic flux analysis reveals the impacts of glutamate on the central metabolism of high β-galactosidase-producing .

BACKGROUND

is a popular recombinant protein expression system for its accessibility of efficient gene manipulation and high protein production. Sufficient supply of precursors, energy, and redox cofactors is crucial for high recombinant protein production. In our present work, we found that the addition of glutamate improved the recombinant β-galactosidase (β-gal) production by G1HL.

METHODS

To elucidate the impacts of glutamate on the central metabolism in detail, a combined C-assisted metabolomics and C metabolic flux analysis was conducted based on LC-MS/MS and GC-MS data.

RESULTS

The pool sizes of intracellular amino acids were obviously higher on glucose/glutamate (Glc/Glu). The fluxes in EMP entry reaction and in downstream TCA cycle were 50 and 67% higher on Glc/Glu than on Glc, respectively. While the fluxes in upstream TCA cycle kept almost unaltered, the fluxes in PPP oxidative branch decreased.

CONCLUSION

The addition of glutamate leads to a remarkable change on the central metabolism of high β-galactosidase-producing G1HL. To meet the increased demands of redox cofactors and energy for higher β-galactosidase production on Glc/Glu, G1HL redistributes the fluxes in central metabolism through the inhibitions and/or activation of the enzymes in key nodes together with the energy and redox status.