Table 3 Strains and plasmids used in this study

Strains

E. coli JM109

recA 1, endA 1, gyrA 96, thi-1, hsdR 17, supE 44, relA 1, Δ(lac-proAB)/F’[traD 36, proAB+, lacIq, lacZ ΔM 15]

TransGen Biotech

BSUL03

ΔamyE::(P₄₃::kivd-adh 2), P₄₃::ilvD-ilvC-alsS; Spc^r, Em^r

[6]

BSUL04

BSUL03 with lactate dehydrogenase inactivation (Δldh); Spc^r, Em^r, Km^r

This study

BSUL05

BSUL04 with pyruvate dehydrogenase complex E2 subunit inactivation (Δldh ΔpdhC); Spc^r, Em^r, Km^r, Tet^r

This study

Plasmids

pUC18

E. coli cloning vector; Amp^r

Laboratory stock

pDK

B. subtilis integration vector with amyE locus; Amp^r, Km^r

BGSC [50]

pHY300PLK

E. coli-B. subtilis cloning vector; Amp^r, Tet^r

Takara

pUCL

pUC18 containing ldh fragment of B. subtilis, Amp^r

This study

pUCLKm

pUCL containing kanamycin resistance cassettes from pDK; Amp^r, Km^r

This study

pUCP01

pUC18 containing homology arm pdhC 1 fragment of B. subtilis; Amp^r

This study

pUCP02

pUCP01 containing homology arm pdhC 2 fragment of B. subtilis; Amp^r

This study

pUCPTet

pUCP02 containing tetracycline resistance cassettes from pHY300PLK; Amp^r, Tet^r

This study