Corynebacterium glutamicum and application thereof

A Corynebacterium glutamicum and glutamine technology, applied in the field of microorganisms, can solve the problems of large-scale industrial production of L-glutamine and low production of L-glutamine, so as to increase the yield and improve the synthesis capacity Effect

Active Publication Date: 2013-10-30
MEIHUA BIOTECH LANGFANG CO LTD
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  • Abstract
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  • Claims
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Problems solved by technology

Wherein Corynebacterium glutamicum is one of bacterial species commonly used in L-glutamine fermentation, but

Method used

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  • Corynebacterium glutamicum and application thereof

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Embodiment 1

[0041] Embodiment 1: the screening of Corynebacterium glutamicum MHZ-0510, MHZ-0511 of the present invention

[0042] The starting strain - wild type Corynebacterium glutamicum B498 was inoculated in LB liquid medium and cultivated at 30°C for 10 hours to obtain a bacterial suspension in the logarithmic growth phase, and then spread the bacterial suspension on the LB solid plate. Appropriate amount of N-methyl-N'nitro-N-nitrosoguanidine solid particles were placed in the center of the LB solid plate, placed in an incubator at 30°C for 16 hours and then scraped to remove N-methyl-N'nitro- N-nitrosoguanidine particles are surrounded by a weak lawn of bacteria suspended in an appropriate amount of sterile saline. Spread the normal saline with bacterial lawn suspended on the LB solid plate containing 1.0 mg / L streptomycin or neomycin respectively, continue to culture at 30°C for 2 days, and pick a single colony according to the characteristics of the colony shape, color, etc., amo...

Embodiment 2

[0046] Embodiment 2: L-glutamine production method of the present invention and L-glutamine fermentation comparative test

[0047] 1. Fermentation method

[0048] Corynebacterium glutamicum MHZ-0510 and MHZ-0511 described in the present invention were respectively inoculated in the seed medium described below for propagation, cultured with shaking at 30°C for 16 hours, and the resulting culture was inoculated into the fermentation described below In the culture medium, the volume ratio of the culture medium to the fermentation medium is 1:10 (those skilled in the art can adjust it according to the amount of the fermentation medium in actual production, this ratio is only the preferred ratio), shake culture at 30°C until no longer Until L-glutamine is produced (generally about 72 hours). During shaking culture in the fermentation medium, maintain the pH value in the fermentation medium between 6.5-7.4.

[0049] Seed medium: 20g / L glucose, 0.5g / L potassium dihydrogen phosphate...

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Abstract

The invention relates to the microbial field, and discloses corynebacterium glutamicum strains and application thereof. A corynebacterium glutamicum MHZ-0510 strain has a preservation number CGMCC No.7939; a corynebacterium glutamicum MHZ-0511 strain has a preservation number CGMCC No.7940; the two strains have significantly improved ability of fermentation for generating L-glutamine, compared with that of the original strain of wild type of corynebacterium glutamicum B498, and can be widely applied to fermentation production of L-glutamine. The corynebacterium glutamicum strains involved in the invention are obtained by screening aminoglycoside antibiotics, such as streptomycin and neomycin, have resistance to the aminoglycoside antibiotics including streptomycin and neomycin, and have improved ability to synthesis of L-glutamine, thus increasing the yield of L-glutamine. Therefore, aminoglycoside antibiotics can be widely applied to screen of L-glutamine.

Description

technical field [0001] The invention relates to the field of microorganisms, in particular to Corynebacterium glutamicum and its application. Background technique [0002] L-Glutamine, the scientific name is 2-amino-5-carboxypentanamide, is the amide of glutamic acid. The molecular formula of L-glutamine is C 10 h 20 N 4 o 6 , the molecular weight is 292.289, and the structural formula is [0003] [0004] L-Glutamine is one of the most abundant amino acids in body fluids, accounting for 61% of the free amino acids in the human body. It is the most common and indispensable conditional essential amino acid in the human body. L-glutamine in the human body is mainly stored in skeletal muscle, and its concentration is 30 times higher than that of the internal environment, and it is released when needed. L-Glutamine has multiple effects, such as enhancing immune function, maintaining acid-base balance, increasing cell volume, enhancing protein synthesis in muscle cells, ...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P13/14C12R1/15
Inventor 高云飞周希贵李岩龚华
Owner MEIHUA BIOTECH LANGFANG CO LTD
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