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Corynebacterium glutamicum for high production of L-ornithine and application thereof

A technology of Corynebacterium glutamicum and ornithine, which can be applied to bacteria, microorganism-based methods, microorganisms, etc., can solve problems such as increased yield, unfavorable industrialized mass output, and poor L-ornithine yield.

Active Publication Date: 2019-01-08
NANJING TECH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

From this result, it can be seen that the strain has high yield at low glucose concentration, but the yield does not increase year-on-year at high glucose concentration, and the yield of L-ornithine per unit time is not good, which is not conducive to large-scale industrial production

Method used

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  • Corynebacterium glutamicum for high production of L-ornithine and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Mutation screening of embodiment 1 Corynebacterium glutamicum (corynbacterium glutamicum) Best-1

[0043] The National Culture Collection Center purchased Corynebacterium glutamicum ATCC 13032, and used liquid LB medium for basic culture. After culturing at 30°C for 1 day, the biomass and the production of L-arginine and L-ornithine were simultaneously measured. cultured in LB seed medium. Dilute the cultured Corynebacterium glutamicum to a cell number of 10 by the 10-fold dilution method 9 / mL, take 0.1mL of the bacterial solution and spread it evenly on a sterile empty plate, and carry out N + ion beam implantation, N + The ion beam implant dose is (95, 140, 185, 230, 275)×2.6×10 13 N+ / cm 2 , N + The ion beam implantation energy is 15keV. After the irradiation, wash the cells with 1mL sterile water, dilute them according to the 10-fold dilution method, and apply them to the plate culture medium. Arginine methyl ester resistance was added to the culture medium, a...

Embodiment 2

[0044] Example 2 Mutation Screening of Corynebacterium glutamicum (corynbacterium glutamicum) jp-07092

[0045] Use Best-1 as the starting strain, use liquid LB medium for basal culture, after culturing at 30°C for 1 day, measure the biomass and the production of L-arginine and L-ornithine at the same time, and culture in liquid LB seed medium . Dilute the cultured Corynebacterium glutamicum to a cell number of 10 by the 10-fold dilution method 9 / mL, take 0.1mL of the bacterial solution and spread it evenly on a sterile empty plate, and carry out N + ion beam implantation, N +The ion beam implant dose is (95, 140, 185, 230, 275)×2.6×10 13 N+ / cm 2 , N + The ion beam implantation energy is 15keV. Wash the cells with 1mL sterile water after the irradiation, dilute them by 10 times and spread them on the plate medium, plate medium (g / L): glucose 5, (NH 4 ) 2 SO 4 3,K 2 HPO 4 0.2, KH 2 PO 4 0.2, MgSO 4 0.5, NaH 2 PO 4 0.2. In addition to the above basic medium,...

Embodiment 3

[0046] Embodiment 3 Cultivation and physiological characteristics of Corynebacterium glutamicum (corynbacterium glutamicum) jp-07092

[0047] The strains are cultured in an aerobic way. The carbon source for cultivating the bacterial strain can be glucose; the nitrogen source for cultivating the bacterial strain is ammonium sulfate. The optimal temperature for the growth of the strain is 25-50°C, the pH range is 5-9, and the optimal pH range is 6-8. Dipotassium hydrogen phosphate, potassium dihydrogen phosphate, magnesium sulfate, manganese sulfate, ferrous sulfate and other inorganic salts can also be added during the strain cultivation process.

[0048] The physiological morphological characteristics of the Corynebacterium glutamicum (corynbacterium glutamicum) jp-07092 are:

[0049] Corynebacterium glutamicum grows vigorously on the LB plate, and it is obvious that the single colony is evenly colored, round or spherical, and off-white in color. The surface of the colony ...

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Abstract

The invention belongs to the technical field of biological fermentation and discloses corynebacterium glutamicum for high production of L-ornithine and application thereof. The corynebacterium glutamicum is classified and named as corynebacterium glutamicum jp-07092 and is preserved in the China General Microbiological Culture Collection Center (CGMCC), and the preservation number is CGMCC No. 15856. The L-ornithine is produced by utilizing strain fermentation, and the corynebacterium glutamicum has the advantages of being low in cost, short in fermentation period, high in unit time yield, simple in subsequent purification process and the like and is more suitable for industrial popularization.

Description

technical field [0001] The invention belongs to the technical field of biological fermentation, and in particular relates to a high-yield L-ornithine Corynebacterium glutamicum and its application. Background technique [0002] Corynebacterium glutamicum (corynbacterium glutamicum) is a very important production strain in the fermentation industry at present. It has been widely used in the production of various amino acids and their related derivatives. Its main products include L-glutamic acid, L -Lysine, L-ornithine, L-glutamine, etc. The fermentative production of amino acids by corynebacterium glutamicum is an aerobic process that produces a large amount of biomass. Corynebacterium glutamicum grows rapidly, has simple culture conditions, strong tolerance, and mature fermentation conditions. Therefore, Corynebacterium glutamicum has become a model strain used in laboratories and industries. [0003] L-ornithine mainly participates in the urea cycle in the living body a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P13/10C12R1/15
CPCC12N1/20C12P13/10C12R2001/15C12N1/205
Inventor 郝宁冯宇静黄彤舸郭格格刘兆星单咸旸
Owner NANJING TECH UNIV
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