TABLE 1

Strains and plasmids used in this study

Strain or plasmidFunction and relevant characteristic(s)Source or reference
Strains
    E. coli DH5αGeneral cloning host [F thi-1 endA1 hsdR17(rK mK) supE44 ΔlacU169 λ ϕ80dlacZΔM15 recA1 gyrA96 relA1]35
    C. glutamicum
        ATCC 13032WT strain, auxotrophic for biotinThis work
        ΔgapA strainIn-frame deletion of the gapA gene of the WT strainThis work
        ΔgapAB strainIn-frame deletion of the gapA and gapB genes of the WT strainThis work
        Δndh strainIn-frame deletion of the ndh gene of the WT strainThis work
        GSM0In-frame deletion of the gapA and gapB genes of the WT strain with pEKEx3-gapNCac overexpressionThis work
    GSM1/2/3/4GSM0 strain with evolved ndh (D213G)This work
Plasmids
    pEKEx3Specr; C. glutamicum-E. coli shuttle vector (Ptac lacIq; pBL1 oriVCg oriVEc)48
    pEKEx3-gapACgDerived from pEKEx3, for regulated expression of C. glutamicum gapA43
    pEKEx3-gapBCgDerived from pEKEx3, for regulated expression of C. glutamicum gapBThis work
    pEKEx3-gapNCacDerived from pEKEx3, for regulated expression of Clostridium acetobutylicum gapNThis work
    pVWEx1Kanr Ptac lacIqThis work
    pVWEx1-udhAEcDerived from pVWEx1, for regulated expression of E. coli udhAThis work
    pVWEx1-ndhCgDerived from pVWEx1, for regulated expression of C. glutamicum ndhThis work
    pVWEx1-ndhD213GDerived from pVWEx1, for regulated expression of C. glutamicum ndhThis work
    pVWEx1-ndhD213NDerived from pVWEx1, for regulated expression of C. glutamicum ndhThis work
    pVWEx1-ndhD213QDerived from pVWEx1, for regulated expression of C. glutamicum ndhThis work
    pK19mobsacBKmr; E. coli-C. glutamicum shuttle vector for construction of insertion and deletion mutants in C. glutamicum (pK18 oriVEc sacB lacZα)This work
    pK19mobsacBΔgapApK19mobsacB with a gapA deletion constructThis work
    pK19mobsacBΔgapBpK19mobsacB with a gapB deletion constructThis work