Microorganism of genus corynebacterium for producing l-arginine, and l-arginine production method using same
A technology of coryneform bacteria and microorganisms, which is applied in the field of microorganisms of the genus Corynebacterium, and can solve the problems of limitation, high-yield L-arginine production and application, etc.
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0037] Example 1: Construction of vectors with enhanced arginine operon
[0038] In order to enhance the arginine operon on the chromosome of microorganisms, a vector was constructed in which the self-promoter of N-acetylglutamyl phosphate reductase (ArgC) was deleted and replaced with a different promoter. As a substitution promoter, lysCP1 (SEQ ID NO: 18 disclosed in Korean Patent No. 10-0930203), which has a strong expression-inducing activity, was used.
[0039] First, the chromosomal DNA of the wild-type strain of Corynebacterium glutamicum (Accession No.: ATCC13869) was used as a template, and a primer pair of SEQ ID NO: 13 (SF_pargC_PR_pDC fusion primer; 5'-CGAGCTCGGTACCCGGGCAAAGAATACGGCTTCCTTGGC-3') and SEQ ID NO: 14 (SR_pargC_PR_XbaI-XhoI-BamHI fusion / restriction enzyme primer; 5'-CTGGATCCTCGAGTCTAGAGACGGGTTAGACATGCAAAA-3') and primer pair SEQ ID NO: 15 (SF_pargC_PR_SpeI-ScaI-BamHI fusion / restriction enzyme primer; 5'-GACTCGAGGATCCAGTACTAGTATGATAATCAAGGTTGCAAT-3') an...
Embodiment 2
[0042] Example 2: Construction of a vector with enhanced ornithine carbamoyltransferase
[0043] To enhance ornithine carbamoyltransferase, one of the arginine biosynthesis enzymes, a recombinant expression vector was constructed. Using p117-cj7-GFP (Korean Patent No. 10-0620092) as a template vector, and removing the nucleotide sequence encoding GFP in the template vector by treatment with EcoRV-Xba I restriction enzyme, and inserting a source derived from Corynebacterium glutamicum Wild-type strains argF and argF2 of ATCC13869 (Korean Patent No. 10-0830290).
[0044] Chromosomal DNA of a wild-type strain of Corynebacterium glutamicum (Accession No.: ATCC13869) was used as a template, and a primer pair of SEQ ID NO: 7 (SF_argF_EcoRV fusion primer; 5'-ACGAAAGGAAACACTCGATATCATGACTTCACAACCACAGGT-3') and SEQ ID NO: 8 ( SR_argF_XbaI fusion primer; 5'-GCCAAAACAGCTCTAGATTACCTCGGCTGGTGGGCCA-3'), the DNA fragment of argF gene was amplified by PCR. The PCR reaction was performed by...
Embodiment 3
[0047] Example 3: Construction of strains with recombinant vectors inserted therein
[0048] 3-1. Insertion of a vector with an enhanced arginine operon
[0049] In order to replace the self-promoter pD-PargC::lysCP1 of the arginine operon on the chromosome of Corynebacterium, the recombinant vector constructed in Example 1 was transformed into an existing strain of arginine-producing Corynebacterium, thereby constructing an insert with Corynebacterium strains of recombinant vectors. Specifically, by transforming the recombinant vector pD-PargC::lysCP1 constructed in Example 1 into the existing arginine-producing strains of KCCM10741P (Korean Patent No. 10-07916590) and ATCC21831, the lysCP1 promoter sequence was inserted into the chromosome , thereby replacing the self-promoter sequence possessed by the parent strain with the promoter sequence of the vector by homologous recombination.
[0050] In the transformation, the recombinant vector was first inserted into KCCM10...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More