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Method for fermenting, extracting and purifying threonine

A technology of threonine and fermentation broth, applied in the field of threonine production, can solve the problems of high mortality rate of strains, waste, and no mother liquor extraction, etc., and achieve the effects of increasing bacterial protein yield, reducing environmental protection pressure, and improving yield.

Active Publication Date: 2019-03-19
齐齐哈尔龙江阜丰生物科技有限公司 +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the literature "Control of acetic acid in the fermentation process of L-threonine, Fermentation Science and Technology Newsletter 2012", in the process of producing threonine by E. generation, but the reduction rate is not obvious, and the yield of threonine cannot be greatly increased, and there is no value for industrial promotion
The medium recorded in the applicant's previous patented technology "A Method for the Production of Ultra-Low Moisture Threonine": glucose 80g / L, corn steep liquor 20g / L, ammonium sulfate 2g / L, calcium carbonate 0.75g / L, KH 2 PO 4 0.2g / L, K 2 HPO 4 0.2g / L, NaCl 0.2g / L, pH value 6.5; the content of threonine in the fermentation broth can reach up to 10g / 100ml, but the mortality rate of the strain is high in the late stage of fermentation, and the consumption of glucose is relatively large. The yield of threonine also needs to be improved
The applicant's previous patented technology "a method for producing granular threonine" increased the yield of threonine through the mixed fermentation process, but in the extraction process, the mother liquor was not extracted, resulting in waste

Method used

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  • Method for fermenting, extracting and purifying threonine
  • Method for fermenting, extracting and purifying threonine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] A method for fermenting, extracting and purifying threonine, comprising the steps of:

[0028] Step 1) Fermentation: Escherichia coli engineering bacteria K12△dapA seed solution (the concentration of the seed solution is 1×10 8 cfu / mL) according to 10% of the inoculum amount into a fermenter containing fermentation medium for fermentation, the temperature is 30°C, the tank pressure is 0.04MPa, the ventilation rate is 0.5vvm, the rotation speed is 100rpm, and the fermentation time is 36h, then Inject Chlamydomonas reinhardtii according to 10% inoculum (the concentration of Chlamydomonas reinhardtii is 1×10 5 cfu / mL), continue to ferment for 48 hours, stop the fermentation, and collect the fermented liquid;

[0029] The components of the fermentation medium are: glucose 20g / L, glycerol 20g / L, corn steep liquor 20g / L, ammonium sulfate 2g / L, potassium dihydrogen phosphate 0.2g / L, dipotassium hydrogen phosphate 0.2g / L, Magnesium sulfate heptahydrate 0.1g / L, ferrous sulfate...

Embodiment 2

[0038] A method for fermenting, extracting and purifying threonine, comprising the steps of:

[0039] Step 1) Fermentation: Escherichia coli engineering bacteria K12△dapA seed solution (the concentration of the seed solution is 1×10 8 cfu / mL) according to the inoculation amount of 6%, was inserted into the fermenter containing the fermentation medium for fermentation, the temperature was 32°C, the tank pressure was 0.04MPa, the ventilation rate was 0.4vvm, the rotation speed was 100rpm, and the fermentation time was 48h. Then insert Chlamydomonas reinhardtii (the concentration of Chlamydomonas reinhardtii is 1×10 5 cfu / mL), continue to ferment for 48 hours, stop the fermentation, and collect the fermented liquid;

[0040] The components of the fermentation medium are: glucose 30g / L, glycerol 30g / L, corn steep liquor 30g / L, ammonium sulfate 3g / L, potassium dihydrogen phosphate 0.3g / L, dipotassium hydrogen phosphate 0.3g / L, Magnesium sulfate heptahydrate 0.2g / L, ferrous sulfat...

Embodiment 3

[0049] The impact of different factors on threonine output and acetic acid output in the technique of the present invention:

[0050] Set group:

[0051] Experimental group: Example 1;

[0052] Control group 1: without adding Chlamydomonas reinhardtii, the rest are the same as in Example 1;

[0053] Control group 2: Glycerin was replaced by glucose of the same quality, and the rest were the same as in Example 1;

[0054] Control group 3: No Chlamydomonas reinhardtii was added, and glycerol was replaced by glucose of the same quality, and the rest were the same as in Example 1.

[0055] The contents of threonine and acetic acid in the final fermentation broth of each group are shown in Table 1:

[0056] Table 1

[0057] group

Threonine g / L

Acetic acid g / L

test group

137.1

0.6

Control group 1

102.5

13.9

Control group 2

118.2

4.7

Control group 3

96.4

15.6

[0058] Conclusion: The experimental group can...

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Abstract

The invention belongs to the technical field of production of threonine and discloses a method for fermenting, extracting and purifying threonine. The method comprises the following steps: (1) fermenting; (2) separating fermenting liquid; (3) carrying out concentration and crystallization, (4) separating crystals; (5) drying; (6) carrying out primary mother solution crystallization and separation;(7) carrying out chromatographic separation; and (8) carrying out crystallization and separation. Threonine fermenting liquid and a waste mother solution are comprehensively utilized by virtue of anextraction and purification process, so that the yield and purity of threonine are increased, the production benefit is achieved, and the environmental protection pressure is alleviated.

Description

technical field [0001] The invention belongs to the technical field of threonine production, and in particular relates to a method for fermenting, extracting and purifying threonine. Background technique [0002] L-threonine (hereinafter referred to as threonine) is a white crystal or crystalline powder, a polar amino acid, odorless and slightly sweet. It melts and decomposes at 253°C. Soluble in water at high temperature, the solubility at 25°C is 20.5g / 100ml, insoluble in ethanol, ether and chloroform. The specific optical rotation is 27.6°-29°, and the isoelectric point is PI5.64. [0003] The threonine industry developed earlier in foreign countries. The United States, France, Germany, Japan and other countries have carried out a series of researches on the synthesis, separation and application of threonine. Ajinomoto Corporation is the largest threonine manufacturer in the world. The threonine produced by the company occupies more than 60% of the global market. The t...

Claims

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Application Information

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IPC IPC(8): C12P13/08C07C227/40C07C227/42C07C229/22C12R1/19C12R1/89
CPCC07C227/40C07C227/42C12P13/08C07C229/22
Inventor 唐永强马延和王健汲广习董力青李航
Owner 齐齐哈尔龙江阜丰生物科技有限公司
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