Equilibrium of the intracellular redox state for improving cell growth and L-lysine yield of Corynebacterium glutamicum by optimal cofactor swapping.

ABSTRACT:

Background

NAD(H/⁺) and NADP(H/⁺) are the most important redox cofactors in bacteria. However, the intracellular redox balance is in advantage of the cell growth and production of NAD(P)H-dependent products.

Results

In this paper, we rationally engineered glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and isocitrate dehydrogenase (IDH) to switch the nucleotide-cofactor specificity resulting in an increase in final titer [from 85.6 to 121.4 g L^-1] and carbon yield [from 0.33 to 0.46 g (g glucose)^-1] of L-lysine in strain RGI in fed-batch fermentation. To do this, we firstly analyzed the production performance of original strain JL-6, indicating that the imbalance of intracellular redox was the limiting factor for L-lysine production. Subsequently, we modified the native GAPDH and indicated that recombinant strain RG with nonnative NADP-GAPDH dramatically changed the intracellular levels of NADH and NADPH. However, L-lysine production did not significantly increase because cell growth was harmed at low NADH level. Lastly, the nonnative NAD-IDH was introduced in strain RG to increase the NADH availability and to equilibrate the intracellular redox. The resulted strain RGI showed the stable ratio of NADPH/NADH at about 1.00, which in turn improved cell growth (?_max.?=?0.31 h^-1) and L-lysine productivity (q_{Lys, max.}?=?0.53 g g^-1 h^-1) as compared with strain RG (?_max.?=?0.14 h^-1 and q_{Lys, max.}?=?0.42 g g^-1 h^-1).

Conclusions

This is the first report of balancing the intracellular redox state by switching the nucleotide-cofactor specificity of GAPDH and IDH, thereby improving cell growth and L-lysine production.

SUBMITTER: Xu JZ 

PROVIDER: S-EPMC6448238 | biostudies-literature | 2019 Apr

REPOSITORIES: biostudies-literature