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Strain for producing L-lysine and method thereof for producing L-lysine

A lysine and strain technology, applied in the biological field, can solve the problems of residual catalyst, limited source of raw materials, poor product safety, etc., and achieve the effects of good genetic stability and increased L-lysine content

Active Publication Date: 2013-10-23
NANJING UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the traditional extraction process is simple, the source of raw materials such as animal blood meal is limited, which is limited to small-scale production; the production of lysine by the synthetic method requires the use of highly toxic raw material phosgene, and there may be residual catalysts, which leads to poor product safety and serious environmental problems ; The chemically synthesized DL-lysine also needs to be split or converted by enzymatic methods to obtain L-lysine, which greatly increases the production cost; microbial fermentation is the most important method for industrial production of lysine at present. Widely available and inexpensive, such as starch (tapioca, cornstarch, etc.), molasses (cane molasses, beet molasses, etc.)

Method used

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  • Strain for producing L-lysine and method thereof for producing L-lysine
  • Strain for producing L-lysine and method thereof for producing L-lysine

Examples

Experimental program
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Effect test

Embodiment 1

[0059] Embodiment 1: This embodiment description will E. coli A301 mutagenesis screening of Escherichia coli as a starting strain NT1003△Met△Thr Methods.

[0060] In this example:

[0061] 1) LB medium, composed of the following mass percentage components: tryptone 1%, yeast extract 0.5%, sodium chloride 1%, the rest is water, pH 6.8-7.2.

[0062] 2) Seed medium, composed of the following mass percentage components: glucose 0.05%, sucrose 0.15%, peptone 3%, yeast extract 3%, ammonium sulfate 2%, dipotassium hydrogen phosphate 0.3%, magnesium sulfate 0.1%, sulfuric acid Ferrous 0.05%, manganese sulfate 0.03%, sodium dihydrogen phosphate 0.25%, calcium carbonate 0.17%, the rest is water, pH 6.8-7.2.

[0063] 3) Fermentation medium, composed of the following mass percentage components: glucose 6%, sucrose 14%, corn steep liquor 1%, peanut cake powder 0.4%, soybean cake powder 0.4%, ammonium sulfate 0.2%, dipotassium hydrogen phosphate 0.08% , magnesium sulfate 0.02%, fe...

Embodiment 2

[0078] According to the mutagenesis screening method in Example 1, Escherichia coli NT1003ΔMetΔThr was obtained.

[0079] Among them, the amount of lithium chloride added was 3% of the mass of the LB plate medium; the ultraviolet irradiation time was 110s; the low-energy nitrogen ion implantation condition was 4 Kv / cm×3 min, and the implantation dose was 50×10 14 ions / cm 2. .

Embodiment 3

[0081] Escherichia coli NT 1003 was obtained according to the mutagenesis screening method in Example 1.

[0082] Among them, the amount of lithium chloride added was 5% of the mass of the LB plate medium; the ultraviolet irradiation time was 30s; the low-energy nitrogen ion implantation condition was 42 Kv / cm×9 min, and the implantation dose was 20×10 14 ions / cm 2. .

[0083] The morphological and physiological and biochemical characteristics of the Escherichia coli NT 1003 obtained in the above-mentioned examples 1-3 are as follows:

[0084] Colony color: off-white.

[0085] Aerobic mode: aerobic growth.

[0086] Colony size: 0.3~0.8×1~4μm.

[0087] Suitable growth temperature: 34-40°C.

[0088] Suitable growth pH: 6.8-7.2.

[0089] Bacteria morphology: round colony with metallic luster.

[0090] Gram stain: negative.

[0091] The genetic stability test of the Escherichia coli NT1003ΔMetΔThr of embodiment 1-3

[0092] In the fermentation medium with glucose and s...

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Abstract

The invention relates to a strain for producing L-lysine, which is classified and named as Escherichiacoli NT1003 delta Met delta Thr and is collected in China Center for Type Culture Collection (CCTCC) on May 30, 2013, and the collection number is CCTCC NO: M2013239. The invention further provides a mutation screening method of the strain and a method for producing the L-lysine by utilizing the strain. After continuous passage is performed on the Escherichiacoli NT1003 delta Met delta Thr strain for 7 times, the content of the L-lysine in a fermentation solution is about 60g / L and has no significant change, so that the genetic stability is good.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to an L-lysine-producing bacterial strain and a method for producing L-lysine. Background technique [0002] L-Lysine (L-Lysine, Lys) is one of the essential amino acids for organisms to maintain life activities. It is the second largest amino acid species in the world after monosodium glutamate, and it is also an important precursor for new materials such as bio-based nylon. The production methods of L-lysine include extraction method, chemical synthesis / enzyme method and microbial fermentation method. Although the traditional extraction process is simple, the source of raw materials such as animal blood meal is limited, which is limited to small-scale production; the production of lysine by the synthetic method requires the use of highly toxic raw material phosgene, and there may be residual catalysts, which leads to poor product safety and serious environmental problems ; The chemic...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N15/01C12N13/00C12P13/08C12R1/19
Inventor 陈可泉何珣应晗笑王震袁佩佩欧阳平凯
Owner NANJING UNIV OF TECH
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