Table 3 Strains and plasmids used in this study

Strain/plasmid

E. coli DH5α

Host for plasmid construction

Lab stock

B. subtilis 168

The bdhA gene donor

Lab stock

E. coli W1485

The udhA gene donor

Lab stock

CGS9

ATCC13032∆pta∆ack∆ldh∆buta∆nagD∆ppc, ∆ackA::P_tuf -alsSD, ∆butA::P_tuf -alsSD, ∆nagD::P_tuf -alsSD, P₁-gltA

[6]

CGK1

CGS9 ∆ldh::Ptrc-bdhA

This study

CGK2

CGS9 ∆ldh::Psod-bdhA

This study

CGK3

CGS9 ∆ldh::Psod -bdhA-udhA

This study

CGK4

CGK3 atpG^{T817C, C818T}

This study

CGK5

CGK4 pECK1

This study

Plasmids

pD-sacB

Kan^R; vector for in-frame deletion (sacB_B.sub.; lacZα; OriV_E.c.)

Lab stock

pD-sacB-ldh

Kan^R; pD-sacB carrying the flanking sequences of the ldh gene

[34]

pD-ldh-A1

Kan^R containing P_trc-bdhA flanks

This study

pD-ldh-A2

Kan^R containing P_sod-bdhA flanks

This study

pD-ldh-AU

Kan^R containing P_sod-bdhA-udhA flanks

This study

pD-sacB-atpG^{T817C, C818T}

KanR, containing the sequence for atpG exchange T817C, C818T

This study

pEC-XK99E

Kan^R; C. glutamicum/E. coli shuttle vector (P_trc, lacIq; pGA1, OriV_C.g., OriV_E.c.)

Lab stock

pECK1

derived from pEC-XK99E with lacIq deleted, for the overexpression of bdhA and udhA under the control of the promoter P_sod and rbs designed for bdhA

This study