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A kind of method for improving the output of l-methionine

A technology of methionine and yield, applied in the field of fermentation, can solve problems such as application limitation, limitation of large-scale chemical production of methionine, and environmental pollution

Active Publication Date: 2020-10-30
ZHEJIANG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the production method of methionine is mainly chemical method, and the D-type and L-type mixed methionine are synthesized, so its application is limited.
The chemical production process of methionine also involves dangerous chemicals such as acrolein, methyl mercaptan, ammonia, and cyanide, and is easy to cause environmental pollution, so it also limits the large-scale chemical production of methionine

Method used

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  • A kind of method for improving the output of l-methionine
  • A kind of method for improving the output of l-methionine
  • A kind of method for improving the output of l-methionine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Cultivation of recombinant Escherichia coli W3110ΔIJAHFEBC / pAm

[0040] (1) Expansion of training:

[0041] Pick a single recombinant E.coli W3110 colony from the kan-resistant LB plate medium and insert it into the 50mg / L kan seed medium, and culture it at 37°C for 12 hours on a shaker at a speed of 180r / min to obtain a seed solution The composition of the seed medium is as follows: peptone 10g / L, NaCl 10g / L, yeast extract 5g / L, solvent is water, pH is natural, sterilized at 115°C for 30min;

[0042] (2) Fermentation culture:

[0043] Inoculate the seed liquid cultivated for 12 hours with a volume concentration of 10% containing 10g / L CaCO 3 and 50mg / L kan in the original fermentation medium, cultured and fermented to OD under the conditions of 28°C and shaker speed of 180r / min 600 =0.6~1.0, add IPTG with a final concentration of 0.1 mM, and continue to cultivate until 48 hours later, the fermentation ends. 10g / L CaCO added to the fermentation medium 3 Control pH5...

Embodiment 2

[0051] In order to increase the production of L-methionine, the cofactor VB was experimentally investigated 12 and its concentration on the production of L-methionine, investigate different VB 12 The effect of concentration on the production of L-methionine.

[0052] Add VB in the original fermentation medium of embodiment 1 step (2) 12 , where VB 12 The final concentrations were 0.5, 1.0, 1.5, 2.0, 2.5 mg / L respectively. Taking no addition as a control, the same conditions were used to carry out fermentation culture, and the content of L-methionine in the fermentation broth was measured after 48 hours of fermentation. The results were as follows: figure 1 .

[0053] From the summary of the experimental results, it can be obtained that by adding VB 12 Can effectively improve the synthesis of L-methionine, but VB 12 The amount should not be too much, too much will be wasted, and it will not be fully utilized in metabolism, which is caused by figure 1 It can be seen that i...

Embodiment 3

[0055] Since L-methionine metabolism involves the metabolism of multiple modules, from carbon skeleton, one-carbon unit and sulfur pathway, only VB 12 A cofactor is not as good as metabolic needs, consider VB from the cofactor 1 Can promote the catabolism of some organic acids in the pathway, thereby increasing the production of L-methionine, so the experiment investigated VB 1 And its concentration L-methionine production.

[0056] The original fermentation medium of embodiment 2 is increased VB 12 cofactor VB 1 , VB 1The concentration of VB is 2, 4, 8, 10, 15 mg / L, and the fermentation reaction is carried out under the same conditions, and different VB 1 Concentration is on the influence of L-methionine output and thallus growth, after fermenting 48h, measure the content of L-methionine in the fermented liquid, add VB with example 2 12 , without adding VB 1 For comparison, the results are as figure 2 .

[0057] From the summary of the experimental results, it can be...

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Abstract

The invention discloses a method for increasing the yield of L-methionine, and belongs to the technical field of fermentation. The method includes the steps that recombinant Escherichia coli E. coli W3110 delta IJAHFEBC / pAm is used as a production bacterial strain, inoculated into a fermentation culture medium, and cultured until OD600 is equal to 0.6-1.0, an inducer IPTG is added, culturing continues until fermenting is ended, and fermentation liquor containing L-methionine is obtained, separated and purified, so that L-methionine is obtained; the fermentation culture medium includes a basicculture medium and cofactors, wherein the cofactors include VB12. In the process of fermenting culture, by optimizing the carbon source concentration of the culture medium, adding precursor amino acidand cofactors and optimizing fermenting conditions, the fermenting yield of L-methionine is increased, the various cofactors are added at the same time, and therefore the fermenting yield of L-methionine can be increased remarkably.

Description

technical field [0001] The invention relates to the technical field of fermentation, in particular to a method for improving the production of L-methionine by using recombinant Escherichia coli to ferment. Background technique [0002] Methionine is an important sulfur-containing amino acid and one of the essential amino acids in the human body. Molecular formula is C 5 h 11 NO 2 S, with a relative molecular weight of 149.21, is white flaky crystal or crystalline powder, with a special smell and slightly sweet taste, soluble in water (3.3g / 100mL, 25°C), dilute acid or alkali. Methionine plays an important role in the functional balance of the human body. Lack of methionine can lead to many diseases, such as: liver disease, affecting pregnancy and Parkinson's disease. Methionine has a wide range of uses and is currently widely used in various fields such as medicine, food, and feed. Appropriate addition in corn-soybean meal-based rations for poultry can effectively impro...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P13/12C12R1/19
CPCC12P13/12
Inventor 徐建妙于筱垣郑裕国柳志强张博
Owner ZHEJIANG UNIV OF TECH
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