Cofactor engineering through heterologous expression of an NADH oxidase and its impact on metabolic flux redistribution in Klebsiella pneumoniae

Table 3 Bacterial strains, plasmids, and primers used in this study

Strains, plasmids, or primers	Genotypes, properties, or sequences^a	Sources or references
Strains
Klebsiella pneumoniae CICC10011	Ap^r, parent strain	CICC
K. pneumoniae XZF-308	Ap^r, Cm^r, recombinant strain of CICC10011, harbring pDK7-nox-2	This work
Escherichia coli DH5α	supE44 ∆ lacU169 (φ80 lacZ ∆ M15) hsdR17 recA1 endA1 gyrA96 thi-1 relA1	TaKaRa
Plasmids
pNOX2	pMosBlue carrying a PCR product encoding NADH oxidase of S. pneumoniae	[38]
pUC18	Ap^r, 2686 bp	TaKaRa
pUC18-nox-2	Ap^r, pUC18 derivative carrying a 1380 bp DNA fragment containing the gene of nox-2, 4066 bp	This work
pDK7	Expressing vector, Cm^r, 4800 bp	[39]
pXZF-308	Cm^r, pDK7 derivative carrying a 1380 bp DNA fragment containing the gene of nox-2, 6180 bp	This work
Primers ^b
P1	5^′-CGGGGTACC TAAGGAGGATATACATATGAGTAAAATCGT-3’ (Kpn I)	This work
P2	5^′-CGGCTGCAG TTATTTTTCAGCCGTAAG-3’ (Pst I)	This work

ISSN: 2731-3654