Impact of gene modification of phosphotransferase system on expression of glutamate dehydrogenase protein of Streptococcus suis in Escherichia coli

Figures & data

Table 1. Strains, plasmids and primers used in this study.

Name	Characteristics	Source
Strains
Z. mobilis	ATCC3 l 821	Lab collection
PAR	E. coli MG1655 (wild type)	Lab collection
PARG	Derived from PAR, but ΔptsG	This study
PARGFK	Derived from PARG, and containing pSTV-FK (glf and glk)	This study
PARGPK	Derived from PARG, and containing pSTV-PK (galP and glk)	This study
SSG	E. coli SS2-GDH (E. coli BL21 containing gdhA of S. suis type 2)	[3]
SSGG	Derived from SSG, but ΔptsG	This study
SSGGFK	Derived from SSGG, and containing pSTV-FK (glf and glk)	This study
SSGGPK	Derived from SSGG, and containing pSTV-PK (galP and glk)	This study
Plasmids
pSTV28	Cm^R, cloning vector	TaKaRa
pSTV-FK	pSTV28 inserted with glfZ.mobilis and glkE. coli	This study
pSTV-FK	pSTV28 inserted with galPE. coli and glkE. coli	This study
pKD46	Am^R, λ red-expressing vector	[11]
pKD3	Cm^R, template vector	[11]
pCP20	Am^R, Cm^R, FLP-expressing vector	[11]
Primers
ptsG-P1	5’-ATGTTTAAGAATGCATTTGCTAACCTGCAAAAGGTCG	This study
	GTAAATCGCTGATGCTGCCTTGAGCGATTGTGTAGGCTG
	GAG-3’^a
ptsG-P2	5’-TTAGTGGTTACGGATGTACTCATCCATCTCGGTTTTCA	This study
	GGTTATCGGATTTAGTACTAACGGCTGACATGGGAATTA
	GC-3’^a
ptsG-P3	5’-AGTTGAAACGTGATAGCCGTC-3’	This study
ptsG-P4	5’-AACGTGGAAGGTTCTATCGTCTAC-3’	This study
glf-P1	5’-TGCACTGCAGCCGATACTCGGCGATTGTAAG-3’^b (PstI)	This study
glf-P2	5’-ACATGCATGCAAGGATTCAGCCAAAGCAAGT-3’^b (SphI)	This study
galP-P1	5’-TGCACTGCAGCGGTCAAACAAGGCAATG-3’^b (PstI)	This study
galP-P2	5’-ACATGCATGCATAGCGGCAGAGGATAGAGC-3’^b (SphI)	This study
glk-P1	5’-CGAGCTCGACTTTAGCGGAGCAGTTG-3’^b (SacI)	This study
glk-P2	5’-TGCACTGCAGAAATCGCCGTTACCAGAC-3’^b (PstI)	This study

^aThe underlines indicate 56-nt homology extensions of a target knockout gene.

^bThe underlines indicate the restriction enzyme cutting site.

Figure 1. Identification of the mutant with deletion of ptsG. M: 1 kb DNA marker (TaKaRa); lane 1: PCR product of ptsG (1900 bp); lane 2: ptsG gene recombinant without chloramphenicol resistance (500 bp).

Figure 2. Effect of the plasmids pSTV-FK and pSTV-PK on cell growth (a) and glucose consumption rate (b) of ptsG mutants.

Figure 3. Effect of modification of the PTS system on the cell density (a) and cell growth rate (b) in expression of GDH protein by E. coli (P < 0.05).

Figure 4. Effect of modification of the PTS system on the concentration (a) and production rate (b) of GDH protein in expression of GDH protein by E. coli (P < 0.05).

Figure 5. Effect of modification of the PTS system on the accumulation of acetate (a) and glucose consumption rate (b) in expression of GDH protein by E. coli (P < 0.05).

Cheng LK, Fan WX, Lei LC, et al. Application of dissolved oxygen control and substrate feeding strategy to improve production of glutamate dehydrogenase protein of Streptococcus suis in Escherichia coli. J Chem Pharm Res. 2016;8(6):223–230.

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