Corynebacterium genus microorganism for producing L-lysine and method for producing L-lysine by using same
A technology of coryneform bacteria and microorganisms, applied to bacteria, introduction of foreign genetic material using vectors, recombinant DNA technology, etc., can solve the problems of unconfirmed changes in L-lysine production, and achieve the effect of enhancing production capacity and high yield
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Embodiment 1
[0037] Example 1: Construction of a recombinant vector for inactivating the odx gene derived from coryneform bacteria
[0038] In order to inactivate the odx gene (SEQ ID NO: 2) encoding the oxaloacetate decarboxylase of Corynebacterium glutamicum having the amino acid sequence represented by SEQ ID NO: 1 by specific gene deletion, a plasmid vector was constructed as follows. First, based on the NIH Genbank of the National Institutes of Health, the base sequence of the odx gene is obtained, and based on the sequence, primers for preparing an inactivated odx gene fragment are synthesized.
[0039] With the chromosome of wild-type Corynebacterium glutamicum ATCC13032 as template, with sequence number 3 and sequence number 4, sequence number 5 and sequence number 6 as primer, carry out PCR (polymerase chain reaction) [Sambrook etc., Molecular Cloning (molecular Cloning), a Laboratory Manual (Experimental Manual) (1989), Cold Spring Harbor Laboratories (Cold Spring Harbor Laborato...
Embodiment 2
[0048] Example 2: Inactivation of odx and analysis of L-lysine productivity in L-lysine-producing coryneform bacteria
[0049] By electric pulse method (Appl.Microbiol.Biotechnol.(Applied Microbiology and Biotechnology) (1999) 52:541-545), the pDZ-Δodx prepared in Example 1 is respectively transformed into as the production L-lysine The strains of Corynebacterium glutamicum KCCM11016P and KCCM11347P, and the transformed strains were obtained on the selection medium containing 25 mg / L kanamycin. The odx gene-inactivated strains were obtained by inserting the DNA fragment Δodx into the genome by secondary recombination (crossover), and these strains were named KCCM11016P / Δodx and KCCM11347P / Δodx, respectively.
[0050] In order to confirm whether the inactivation of the odx gene will actually improve the production capacity of lysine, the odx-inactivated strains KCCM11016P / Δodx and KCCM11347P / Δodx were cultured by the following methods, and their production capacity was confirme...
Embodiment 3
[0061] Example 3: Inactivation of odx and confirmation of lysine-producing ability in L-lysine-producing coryneform bacteria with enhanced L-lysine biosynthetic pathway
[0062] Using the same method as in Example 2, the pDZ-Δodx prepared in Example 1 was transformed into Corynebacterium glutamicum KCCM10770P (Korean Authorized Patent No. 10-0924065) to prepare a transformed strain, which was named KCCM10770P / Δodx.
[0063] In order to confirm whether the inactivation of the odx gene can actually improve the production capacity of lysine, the odx-inactivated strain KCCM10770P / Δodx was cultivated by the same method as in Example 2, and its production capacity was confirmed. The L-lysine content in the culture of each strain is shown in Table 2 below.
[0064] [Table 2]
[0065]
[0066] As shown in Table 2, it could be confirmed that the L-lysine production ability of the odx-inactivated strain KCCM10770P / Δodx was increased by about 6.4% compared with the parent strain KCC...
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