Method for producing L-tryptophan by fermentation

A tryptophan and process technology, applied in the field of amino acid fermentation, can solve the problems of difficult control of fermentation parameters, complex fermentation process, and difficult operation, and achieve the effects of improving cell metabolism pathways, increasing fermentation yield, and increasing fermentation conversion rate

Inactive Publication Date: 2019-04-09
XINJIANG FUFENG BIOTECH +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The present invention adopts separation and separation of three columns in series, decolorization of ultrafiltration membrane and one-time crystallization to obtain L-tryptophan product, the process is simple and continuous production is possible, and protein feed can be produced from process waste after comprehensive treatment, which is green and environment-friendly; the method The method of mixed fermentation improves the efficiency of acid production, but there are problems such as relatively complicated fermentation process and difficult control of fermentation parameters.
In response to the above defects, the applicant's patented technology "CN201711289822" discloses the method of using a single strain of fermentation, adding tourmaline and using dialysis medium to increase the amount of acid produced by fermentation; this method increases the amount of acid produced by fermentation, but needs to replace the culture base, the operation is more difficult, and it is easy to cause the death of some strains

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  • Method for producing L-tryptophan by fermentation

Examples

Experimental program
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Effect test

Embodiment 1

[0029] A process for producing L-tryptophan by fermentation, comprising the steps of:

[0030] Preparation of fermentation medium: glucose 20g / L, dipotassium hydrogen phosphate 9g / L, yeast extract 5g / L, citric acid 4.6g / L, ammonium sulfate 1.8g / L, magnesium sulfate 1.6g / L, choline chloride 0.4 g / L, ferrous sulfate 65mg / L, biotin 0.2mg / L; pH controlled at 6.8, sterilized at 115°C for 15 minutes;

[0031] Prepare feeding nutrient solution: the concentration of glucose is 100g / L, the concentration of indole is 1g / L, the concentration of inositol is 0.5g / L, and sterilized at 115°C for 15min;

[0032] Escherichia coli E. coli TRTH was taken as an example, the seed solution (OD 600 The value is 10.8) into the fermentation medium, the culture temperature is 36.8-37.0 ℃, the dissolved oxygen is 20%, and 25% ammonia water is automatically fed through the numerical control to maintain the pH of the tank system between 7.0-7.2, and the defoamer is added Defoaming; when the fermentati...

Embodiment 2

[0034] A process for producing L-tryptophan by fermentation, comprising the steps of:

[0035] Preparation of fermentation medium: glucose 20g / L, dipotassium hydrogen phosphate 9g / L, yeast extract 5g / L, citric acid 4.6g / L, ammonium sulfate 1.8g / L, magnesium sulfate 1.6g / L, choline chloride 0.5 g / L, ferrous sulfate 65mg / L, biotin 0.2mg / L; pH controlled at 6.6, sterilized at 115°C for 15min;

[0036] Prepare feeding nutrient solution: the concentration of glucose is 100g / L, the concentration of indole is 1.5g / L, the concentration of inositol is 0.8g / L, and sterilized at 115°C for 15min;

[0037] Escherichia coli E. coli TRTH was taken as an example, the seed solution (OD 600 The value is 10.7) into the fermentation medium, the culture temperature is 36.8-37.0 ℃, the dissolved oxygen is 20%, and 25% ammonia water is automatically fed through the numerical control to maintain the pH of the tank system between 7.0-7.2, and the defoamer is added Defoaming; when the fermentation ...

Embodiment 3

[0039] 1. The effect of the added amount of choline chloride on the bacterial biomass and tryptophan production in the fermentation broth.

[0040] The operation method refers to Example 1, and the addition amount of choline chloride is selected to be 0, 0.2, 0.4, 0.6, 0.8, 1 (g / L); Figure 1-2 As shown, with the increase of the amount of choline chloride, the biomass of the bacteria gradually increased. When the concentration of choline chloride was increased to 0.4 g / L, the cell growth was significantly improved, the cells grew rapidly, and the tryptophan The accumulation rate has also been rapidly improved. Continue to increase the amount of choline chloride added, but the bacterial biomass and tryptophan production have not increased significantly. It is more appropriate to choose an added amount of 0.4-0.6g / L.

[0041]2. Effects of indole and inositol on bacterial biomass, tryptophan production, sugar-acid conversion rate and fermentation acid production rate in fermentat...

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Abstract

The invention belongs to the technical field of amino acid production and discloses a method for producing L-tryptophan by fermentation. The method comprises the following steps: an escherichia coli seed solution for producing L-tryptophan is inoculated to a fermentation medium with the inoculum size being 10%, culture temperature is 36.8-37.0 DEG C, dissolved oxygen amount is 20%, and pH in a tank is kept in the range of 7.0-7.2 by fed-batch of ammonium hydroxide; when fermentation is performed for 24 h, a nutrient solution is added to the tank in a fed-batch manner, sugar concentration in the tank is kept to be 0.5-1 g/L until fermentation is ended, the total fermentation time is 42-48 h, and L-tryptophan fermentation broth is obtained. The process is simple, and the fermentation efficiency and the glucose-to-acid conversion rate are high.

Description

technical field [0001] The invention belongs to the technical field of amino acid fermentation, and in particular relates to a process for producing L-tryptophan by fermentation. Background technique [0002] The molecular formula for L-tryptophan is C 11 h 12 o 2 N 2 , with a molecular weight of 204.21 and a nitrogen content of 13.72%. L-Tryptophan is a neutral aromatic amino acid containing indole groups. It is silky luster, hexagonal flaky self-colored crystals, odorless and sweet. Solubility in water is 1.14g / L (25°C), soluble in dilute acid or alkali, relatively stable in lye, decomposed in strong acid, slightly soluble in ethanol, insoluble in chloroform and ether. [0003] L-Tryptophan is one of the eight essential amino acids in human and animal life activities, and it plays an important role in the growth, development and metabolism of humans and animals. It is called the second essential amino acid, following methionine and lysine Amino acid is the third larg...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P13/22C12R1/19
CPCC12P13/227
Inventor 徐庆阳李德衡王健刘元涛包鑫张宗华杨瑞丽边恩来王飞米永花
Owner XINJIANG FUFENG BIOTECH
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