Metabolic engineering of Escherichia coli BW25113 for the production of 5-Aminolevulinic Acid based on CRISPR/Cas9 mediated gene knockout and metabolic pathway modification

Table 2 ALA production in recombinant E. coli expressing various related genes

Strain	Relevant characteristic(s)	Plasmid	Expressed genes	ALA accumulation (mg/L)	Standard Error	Relative Change	ALA production rate (g/g)
E. coli BW25113-T7	BW25113-T7	pET-ALA-LAA	hemA, hemL, eamA	662.25	100.28	1.00	0.066
E. coli XD	BW25113-T7	pET-ALA-LAA + pUC-gltX-gltD	hemA, hemL, eamA, gtlX, gltD	476.37	18.45	0.72	0.048
E. coli BX	BW25113-T7	pET-ALA-LAA + pUC-gltB-gltX	hemA, hemL, eamA, gltB, gltX	503.16	37.35	0.76	0.050
E. coli BD	BW25113-T7	pET-ALA-LAA + pUC-gltB-gltD	hemA, hemL, eamA, gltB, gltD	487.92	46.40	0.74	0.049
E. coli BDX	BW25113-T7	pET-ALA-LAA + pUC-gltB-gltX-gltD	hemA, hemL, eamA, gltB, gltD, gltX	293.65	74.89	0.44	0.029
E. coli D5:FYABD	BW25113-T7 ΔhemF, ΔybdK, ΔgdhA, ΔgadB, ΔdppA	pET-ALA-LAA	hemA, hemL, eamA	1601.72	98.70	2.42	0.160
E. coli D5:FYABD-RSA	BW25113-T7 ΔhemF, ΔybdK, ΔgdhA, ΔgadB, ΔdppA	pET-ALA-LAA + pACYCD-RS-hemA	hemA, hemL, eamA, hemA(RS)	2099.71	159.20	3.17	0.210

A 1% (v/v) inoculum from an overnight culture for 12 h was used. IPTG was added when OD600 reached 0.7. Samples were taken and measured until 24 h. 10 g/L glucose was added initially as sole carbon source. Glycine (2 g/L) as substrate for C4 pathway was added as indicated. The results are the average of three individual experiments. ALA production rate: yield of ALA (g) per g glucose

ISSN: 1754-1611