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Method for producing tryptophan through fermentation of fed-batch culture liquid

A technology of fed-feed culture and tryptophan, applied in the field of microbial fermentation, can solve the problems of unsuitability for large-scale popularization, increased metabolic flow, and high cost, and achieves the effects of smooth energy circulation and transmission, stable growth cycle, and reduced production costs.

Pending Publication Date: 2019-12-06
XINJIANG FUFENG BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The literature "Research on L-tryptophan serum fermentation process, Food and Fermentation Technology, Issue 2, 2017" took the metabolic flow distribution of fermentation broth in the middle and late stages of fermentation as a monitoring point, and studied the effect of serum fermentation medium on the production of L-tryptophan by Escherichia coli TRTH fermentation. -The influence of tryptophan, compared with the traditional ordinary fermentation medium, in the middle and late stages of fermentation, the metabolic flow of L-tryptophan synthesis increases, and the metabolic flow of the main by-product acetic acid decreases obviously; but the cleaning The fermentation medium uses a large number of different types of amino acids, which is expensive and not suitable for large-scale promotion and use

Method used

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  • Method for producing tryptophan through fermentation of fed-batch culture liquid
  • Method for producing tryptophan through fermentation of fed-batch culture liquid
  • Method for producing tryptophan through fermentation of fed-batch culture liquid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] A method for fermented production of tryptophan with fed culture broth, which comprises the following steps:

[0038] Step 1): The L-tryptophan engineered bacteria Escherichia coli ( E. coli ) TRTH, cultured to a certain concentration of bacterial liquid;

[0039] Step 2): Inoculate the bacterial liquid obtained in step 1) with 0.5% inoculum in a shake flask containing a first-level seed culture medium, and cultivate it at 36°C at 100-200 rpm for 10-20 hours;

[0040] Step 3): Inoculate the first-level shake flask seeds obtained in step 2) in the seed tank of the second-level seed culture medium at a 5% inoculum amount, control the air volume 5-10L / min, rotate speed 200-400rpm, and temperature 35-37 ℃, tank pressure 0.05-0.08MPa, pH value controlled at 6.5-6.8 by ammonia water, culture for 10-20h;

[0041] Step 4): The secondary fermentor seed (OD 600 Value is 11) Inoculate in a fermenter equipped with fermentation medium with 7% inoculum, rotate speed 400rpm, temperature 36℃...

Embodiment 2

[0051] A method for fermented production of tryptophan with fed culture broth, which comprises the following steps:

[0052] Step 1): The L-tryptophan engineered bacteria Escherichia coli ( E. coli ) TRTH, cultured to a certain concentration of bacterial liquid;

[0053] Step 2): Inoculate the bacterial liquid obtained in step 1) with 0.1%-1% of the inoculum amount in a shake flask containing a first-level seed culture medium, and cultivate it at 36°C at 100-200rpm for 10-20h;

[0054] Step 3): Inoculate the first-level shake flask seeds obtained in step 2) in the seed tank of the second-level seed culture medium at an inoculum of 1%-10%, control the air volume at 5-10L / min, rotate at 200-400rpm and temperature 35-37℃, tank pressure 0.05-0.08MPa, pH value controlled at 6.5-6.8 by ammonia water, culture for 10-20h;

[0055] Step 4): The secondary fermentor seed (OD 600 Value is 12) Inoculate in a fermenter equipped with fermentation medium with 6% inoculum, rotate at 300rpm, temperat...

Embodiment 3

[0065] 1. The effect of the amount of dipotassium hydrogen phosphate added on the production of L-tryptophan.

[0066] The fermentation process is the same as in Example 1, without the step of culture solution. First, verify the effect of adding potassium hydrogen phosphate solution on fermentation through experiments, such as figure 1 As mentioned, with the increase of the amount of dipotassium hydrogen phosphate, the content of L-tryptophan in the fermentation broth will increase accordingly. When the flow rate is 2ml / h, it basically reaches the peak value. Continue to increase the flow rate, which has no effect on the tryptophan output Significantly higher, the influence trend of dipotassium hydrogen phosphate on bacterial biomass is similar to that of tryptophan (not shown in the figure). The main reason is that the inflow of dipotassium hydrogen phosphate solution can increase the bacterial biomass, thereby increasing the tryptophan The yield.

[0067] 2. The effect of ammoni...

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Abstract

The invention belongs to the technical field of microbial fermentation, and discloses a method for producing tryptophan through fermentation of fed-batch culture liquid. In the culture process of a fermentation tank, the tryptophan is produced through a fed-batch dipotassium phosphate solution or an ammonium sulfate solution or an arginine aqueous solution in a fed-batch mode, and culture is conducted for 40 h and then completed. Under the conditions of not additionally arranging any additional equipment and not increasing any manpower input, the fermentation cycle is prolonged, the labor intensity is lowered, and the yield of the tryptophan is significantly increased.

Description

Technical field [0001] The invention belongs to the technical field of microbial fermentation, and specifically relates to a method for producing tryptophan by fermenting fed-batch culture broth. Background technique [0002] The molecular formula of tryptophan is C 11 H 12 O 2 N 2 , The molecular weight is 204.21, and the nitrogen content is 13.72%. Tryptophan is a neutral aromatic amino acid containing an indolyl group. It has a silky luster, hexagonal flaky white crystals, odorless and sweet. Solubility in water is 1.14g / L (25℃), soluble in dilute acid or alkali, stable in lye, decomposed in strong acid, slightly soluble in ethanol, insoluble in chloroform and ether. [0003] Tryptophan is one of the eight essential amino acids in the life activities of humans and animals. It plays an important role in the growth and metabolism of humans and animals. It is called the second essential amino acid. It is the second essential amino acid after methionine and lysine. The third large...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P13/22C12N1/20C12R1/19
CPCC12P13/227C12N1/20
Inventor 包鑫冯世红张宗华崔小红边恩来郭旭辉
Owner XINJIANG FUFENG BIOTECH
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