Engineering of Corynebacterium glutamicum for growth and production of L-ornithine, L-lysine, and lycopene from hexuronic acids

Hadiati, Atika; Krahn, Irene; Lindner, Steffen N; Wendisch, Volker F

doi:10.1186/s40643-014-0025-5

Bioresources and Bioprocessing

Table 1 Strains, plasmids, and oligonucleotides used in this study

From: Engineering of Corynebacterium glutamicum for growth and production of L-ornithine, L-lysine, and lycopene from hexuronic acids

Name	Relevant genotype/information	Reference
*E. coli* strain
DH5α	(ф80lacZΔM15) Δ(lacZYA-argF)U169 deoR recA1 endA1 hsdR17(rK−, mK−) supE44 thi1 gyrA96 relA1	[38]
*C. glutamicum* strains
WT	Wild-type strain ATCC 13032, auxotrophic for biotin	[40]
ORN1	In-frame deletion of argR and argF, L-ornithine overproducing strain derived from C. glutamicum WT	[14]
ORN1	ATCC 13032, auxotrophic for L-arginine	[14]
DM1933	Δpck deletion mutant of C. glutamicum WT also carrying the chromosomal changes pyc(P458S), hom(V59A), two copies of lysC(T311I), two copies of asd, two copies of dapA, two copies of dapB, two copies of ddh, two copies of lysA, two copies of lysE	[37]
ΔcrtYEb	crtY _e Y _f Eb deletion mutant of C. glutamicum MB001	[23]
Plasmids
pEKEx3	Spec^R; E. coli/C. glutamicum shuttle vector for regulated gene expression (P_tac, lacI ^q, pBL1 oriV _Cg)	[41]
pHexA	pEKEx3 derivative for IPTG-inducible expression of uxaCAB, uxuAB, kdgK, and eda from E. coli containing artificial ribosome binding sites each	This work
pVWEx1	Km^R; E. coli/C. glutamicum shuttle vector for regulated gene expression (P_tac, lacI ^q, pCG1 oriV _Cg)	[42]
pVWEx1-exuT	pVWEx1 derivative for IPTG-inducible expression of exuT from E. coli containing artificial ribosome binding site	This work
Primers
G1a	GCAGGTCGACTCTAGAGGATCCCCGAAAGGAGGCCCTTCAGATGACTCCGTTTATGACTGAAGATTTC
G1b	GTACTAGCTAATGCAATCAGTGATGTTATAGCGTTACGCCGCTTTTG
G2d	CATCACTGATTGCATTAGCTAGTACGAAAGGAGGCCCTTCAGATGAAAACACTAAATCGTCGCGAT
G2c	GCTAATGGTGCTATCTGGTACGATCTTAGCACAACGGACGTACAG
G3d	GATCGTACCAGATAGCACCATTAGCGAAAGGAGGCCCTTCAGATGGAACAGACCTGGCGC
G3f	GCAGGTCGACTCTAGAGGATCCCCATGGAACAGACCTGGCGCTGGTACGGCC
G3c	CGTTCTAGTTACTTTGGAACGTACCTTACAGCGCAGCCACACA
G4d	GGTACGTTCCAAAGTAACTAGAACGGAAAGGAGGCCCTTCAGATGTCCAAAAAGATTGCCGTGAT
G4c	GTCAATCCATGGCATTCTAGCCAAGTTACGCTGGCATCGCCTC
G5d	CTTGGCTAGAATGCCATGGATTGACGAAAGGAGGCCCTTCAGATGAAAAACTGGAAAACAAGTGCAG
G5c	GCCAGTGAATTCGAGCTCGGTACCCTTACAGCTTAGCGCCTTCTACAG
ExuT-fw	CTGCAGGTCGACTCTAGAGGAAAGGAGGCCCTTCAGA
ExuT-rv	CGGTACCCGGGGATCTTAATGTTGCGGTGCGGGATC

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