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Method for high-efficient production of L-tryptophan by microbiological fermentation method

A microbial fermentation method and tryptophan technology, applied in the field of efficient production of L-tryptophan by microbial fermentation method, can solve the problems of high cost of enzymatic conversion method, raw material source environment, large amount of pollution, etc., and achieve simple and easy control of the process , good application prospects, and the effect of increasing fermentation yield

Active Publication Date: 2016-08-17
HENAN JULONG BIOLOGICAL ENG CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because chemical synthesis and protein hydrolysis require a large amount of organic solvents, there are problems such as raw material sources and environmental pollution, and are rarely used in production.
Enzymatic conversion method has high cost and large amount of pollution

Method used

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  • Method for high-efficient production of L-tryptophan by microbiological fermentation method
  • Method for high-efficient production of L-tryptophan by microbiological fermentation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] A method for efficiently producing L-tryptophan by microbial fermentation, comprising the following steps:

[0024] (1) Cultivated mature strains

[0025] Preparation of bacteria solution: Take an eggplant-shaped bottle with full growth of bacteria, inject 50ml of physiological saline under flame protection, scrape the surface bacteria into the physiological saline with an inoculation loop or a glass scraper, and then put the physiological The saline is aseptically transferred to the inoculation bottle.

[0026] The strain is Escherichia coli JLTrp, which is classified as Escherichia coli (Escherichia coli), and has been preserved in the General Microbiology Center (CGMCC) of the China Microbiological Culture Collection Management Committee (CGMCC), and the strain preservation number is CGMCC NO. 7.217, the deposit date is May 8, 2016.

[0027] Cultivation of mature seeds:

[0028] a. Empty tank sterilization: clean the seed culture tank, carefully check the valves a...

Embodiment 2

[0043] A method for efficiently producing L-tryptophan by microbial fermentation, comprising the following steps:

[0044] (1) Cultivated mature strains

[0045] Preparation of bacterial solution: Take an eggplant-shaped bottle with full growth of Escherichia coli JLTrp in the bottle, inject 50ml of normal saline under flame protection, and use an inoculation loop or a glass scraper to scrape the surface bacteria into the normal saline, and then put The physiological saline containing the bacteria was transferred to the inoculation bottle under aseptic operation.

[0046] Cultivation of mature seeds:

[0047] a. Sterilization of empty cans: clean the tank body, carefully check the valves and manholes, and then turn on steam to sterilize the empty cans. The sterilization pressure is 0.12MPa, the temperature is 121°C, and the time is 45min;

[0048] b. Sterilization of solid tank: After fully dissolving the liquid seed culture medium, pump it into the seed culture tank with a ...

Embodiment 3

[0062] A method for efficiently producing L-tryptophan by microbial fermentation, comprising the following steps:

[0063] (1) Cultivated mature strains

[0064] Preparation of bacterial solution: Take an eggplant-shaped bottle in which Escherichia coli JLTrp has grown fully, inject 50ml of normal saline under flame protection, and use an inoculation loop or a glass scraper to scrape the surface bacteria into the normal saline, and then Transfer the normal saline containing the bacteria into the inoculation bottle under aseptic operation.

[0065] Cultivation of mature seeds:

[0066] a. Sterilization of empty cans: clean the tank body, carefully check the valves and manholes, and then turn on steam to sterilize the empty cans. The sterilization pressure is 0.12MPa, the temperature is 121°C, and the time is 45min;

[0067] b. Solid tank sterilization: pump the liquid seed culture medium into the seed culture tank, open the tube and heat up to 80°C, then open the steam to ent...

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Abstract

The invention relates to a technique for high-efficient production of L-tryptophan by a microbiological fermentation method. The technique comprises the following steps: (1) culturing a mature Escherichia coli JLTrp strain; (2) culturing a fermentation tank: inoculating the mature strain to the fermentation tank; keeping the temperature to be 34 to 36 DEG C; carrying out supplementary liquid ammonia subsection control on pH, wherein pH is 7.0 to 7.2 in 0 to 18 hours, and 6.4 to 6.8 in 18 hours to the time of tank discharging; carrying out dissolved oxygen subsection control, wherein the dissolved oxygen is 10 to 25% in 0 to 18 hours, and 15 to 40% in 18 hours to the time of discharging the tank, the pressure is 0.03 to 0.08 MPa, and the air volume is 0.3 VVM to 2.0 VVM; controlling residual sugar to be 0.01 to 0.5% in the process, and then culturing to prepare the L-tryptophan. The method for the high-efficient production of the L-tryptophan by the microbiological fermentation method disclosed by the invention aims at good culture medium components and culture conditions, and the concentration control of the residual sugar, temperature control, and pH and dissolved oxygen subsection control are performed in a fermentation process, so that the fermentation period is shortened, the fermentation yield is improved, the process is simple and easy to control, and the cost is relatively low, furthermore, the method for the high-efficient production of the L-tryptophan by the microbiological fermentation method is suitable for large-scale industrial application.

Description

technical field [0001] The invention relates to the technical field of microbial fermentation, in particular to a method for efficiently producing L-tryptophan by microbial fermentation. Background technique [0002] Tryptophan, also known as α-amino-β-indolepropionic acid, has two isomers, D-type and L-type, and only L-tryptophan exists naturally. L-Tryptophan is a neutral aromatic amino acid containing indolyl, scientific name: β-indolylalanine; English name Tryptophan, molecular formula C11H12N2O2, relative molecular weight 204.21, melting point 289°C. L-Tryptophan is white or light yellow crystal or crystalline powder, no odor and slightly bitter. Slightly soluble in water, very slightly soluble in ethanol. It widely exists in natural proteins. L-tryptophan plays an important role in the growth, development and metabolism of human body and animals, and is widely used in medicine, food, feed and other fields. In recent years, with the application of tryptophan in medi...

Claims

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Application Information

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IPC IPC(8): C12P13/22C12R1/19
CPCC12P13/227
Inventor 曹华杰刘帅李静岳贵龙张孟涛
Owner HENAN JULONG BIOLOGICAL ENG CO LTD
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