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Method for fermentation production of L-tryptophan with high efficiency

A tryptophan, high-efficiency technology, applied in the direction of biochemical equipment and methods, fermentation, bioreactor/fermenter combination, etc., can solve the problems of short fermentation cycle, high cost, low fermentation unit, etc., to reduce labor intensity, The effect of improving accuracy and prolonging the fermentation cycle

Active Publication Date: 2014-03-05
SHANDONG LUKANG PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to overcome the problems of low fermentation unit, short fermentation cycle and high cost in the existing production process, and to invent a high-efficiency Method for producing L-tryptophan by fermentation

Method used

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  • Method for fermentation production of L-tryptophan with high efficiency

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Experimental program
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Effect test

Embodiment 1

[0040] Seed culture medium: glucose 5.0%, disodium hydrogen phosphate 0.2%, peptone 1.0%, magnesium chloride 0.1%, trisodium citrate 0.10%, ferrous sulfate 0.001%, manganese sulfate 0.0001%, the above are weight percentages, before sterilization NaOH adjusted the pH to 6.5.

[0041] Fermentation medium: 5.0% glucose, 0.5% disodium hydrogen phosphate, 0.5% peptone, 0.2% citric acid, 0.0009% manganese sulfate, 0.02% ferrous sulfate, the above are percentages by weight, and the pH is adjusted to 6.5, adjust the pH to 6.5 with high-concentration ammonia water after sterilization.

[0042] Insert the strains into the seed medium with an inoculum size of 0.1% v / v, and cultivate them in a 1000L seed tank under the conditions of tank temperature 33°C, pH 6.5, tank pressure 0.06MPa and dissolved oxygen above 60%. ~17h, when the absorbance value at 600nm is 10~30, inoculate 4m with 1.0% v / v 3 In the fermentation tank, the following process is used to control: tank temperature 33°C, ta...

Embodiment 2

[0045] Seed culture medium: glucose 2.0%, disodium hydrogen phosphate 0.2%, peptone 1.0%, magnesium chloride 0.1%, trisodium citrate 0.10%, ferrous sulfate 0.001%, manganese sulfate 0.0001%, the above are weight percentages, before sterilization NaOH adjusted the pH to 7.5.

[0046] Fermentation medium: 0.5% glucose, 0.5% disodium hydrogen phosphate, 0.5% peptone, 0.5% citric acid, 0.0009% manganese sulfate, 0.02% ferrous sulfate, the above are percentages by weight, and the pH is adjusted to 7.5, adjust the pH to 7.5 with high-concentration ammonia water after sterilization.

[0047] Insert the strains into the seed medium with an inoculum size of 5.0% v / v, and cultivate them in a 1000L seed tank under the conditions of tank temperature 38°C, pH 6.5, tank pressure 0.05MPa and dissolved oxygen above 60%. ~17h, when the absorbance value at 600nm is 10~30, inoculate 4m with 15% v / v 3 In the fermenter, the following process is used to control: tank temperature 28°C, tank pressu...

Embodiment 3

[0050] Seed medium: 2.5% glucose, 1.5% disodium hydrogen phosphate, 0.5% peptone, 0.5% magnesium chloride, 0.50% trisodium citrate, 0.0001% ferrous sulfate, 0.001% manganese sulfate, the above are weight percentages, before sterilization NaOH adjusted the pH to 6.8.

[0051] Fermentation medium: 5.0% glucose, 1.5% disodium hydrogen phosphate, 0.15% peptone, 0.2% citric acid, 0.00045% manganese sulfate, 0.002% ferrous sulfate, the above are percentages by weight, and the pH is adjusted to 6.8, adjust the pH to 6.8 with high-concentration ammonia water after sterilization.

[0052] Insert the strains into the seed culture medium, the inoculation amount is 3.0% v / v, and cultivate them in a 1000L seed tank under the conditions of tank temperature 30°C, pH 6.8, tank pressure 0.04MPa and dissolved oxygen above 60%. ~17h, when the absorbance value at 600nm is 10~30, inoculate 4m with 15% v / v 3 In the fermentation tank, the following process is used for control: tank temperature 30°...

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Abstract

The invention discloses a method for fermentation production of L-tryptophan with high efficiency. Through modification of a fermentation device and optimization of a fermentation medium formula and a fermentation technology, the problems of low fermentation unit and short fermentation cycle in existing production technologies are solved. The method comprises steps: a sugar supplementing system of an existing device is modified, and a single pipeline is replaced by a sugar distribution assembly through which sugar is distributed in a fermentation liquid rapidly and uniformly; the ratios of components in fermentation medium are optimized and adjusted; during the fermentation process, dissolved oxygen is controlled at a certain level through adjustment of stirring speed and air flow, the pH value is controlled through fed-batch of high-concentration ammonia water, the residual sugar content is controlled below 0.1% through fed-batch of sugar (comprising glucose and liquid sugar), and the culture is finished after 40-60h. The method prolongs the fermentation cycle, lowers the labor intensity, raises the fermentation unit greatly and lowers the production cost without addition of any extra device and manpower investment.

Description

technical field [0001] The invention relates to a method for producing L-tryptophan, in particular to a method for producing L-amino acid with high yield and efficient fermentation. Background technique [0002] The production method of L-tryptophan has successively experienced three methods: protein hydrolysis, chemical synthesis and microbial method, wherein the microbial method includes direct fermentation method, microbial conversion method and enzymatic method. With the continuous deepening of genomics and metabolomics research, people have applied recombinant DNA technology to microbial breeding and enzyme industry, thus greatly promoting the industrialization process of direct fermentation and enzymatic production of L-tryptophan. The raw materials of the direct fermentation method are simple, low in cost, good in quality, and low in environmental protection treatment costs, and have great advantages. At present, many scientific research institutes and enterprises at...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P13/22C12M1/00
Inventor 徐洪利左良成王文笙张建军赵斐田晓梅宋爱刚慕东
Owner SHANDONG LUKANG PHARMA
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