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High-efficiency production method of gamma-polyglutamic acid

A polyglutamic acid, high-efficiency technology, applied in the field of high-efficiency production of γ-polyglutamic acid, can solve the problems of L-glutamic acid-independent production efficiency, low production efficiency, unsuitable for economical scale production, etc., to improve conversion rate, reducing the effect of remaining

Active Publication Date: 2014-04-02
天津北洋百川生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

L-glutamic acid-independent production efficiency is low, so industrial fermentation production basically uses L-glutamic acid-dependent strains, L-glutamic acid is added to the medium as an important precursor, but its about 50% The conversion rate is not suitable for economical large-scale production, and it is still a need to find a high-efficiency and low-cost γ-polyglutamic acid production method

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] A method for efficient production of gamma-polyglutamic acid, comprising the steps of:

[0022] (1) Strain activation: the original strain of Bacillus licheniformis CGMCC NO.3336 was inoculated on the slant medium, cultured at 37°C for 16 hours, activated twice in this way, and mature slant seeds were prepared;

[0023] The slant medium consists of: peptone 10g / L, yeast powder 5g / L, NaCl10g / L, agar 20g / L, pH7.2, and the insufficient part is supplemented with pure water;

[0024] (2) Preparation of seed solution: inoculate one ring of activated slant seeds into a 500ml Erlenmeyer flask filled with 50ml of liquid medium, and culture at 37°C and 220rpm for 16 hours to logarithmic growth phase;

[0025] The seed medium consists of: glucose 30g / L, yeast extract 7g / L, tryptone 10g / L, K 2 HPO 4 0.5g / L, MgSO 4 ·7H 2 O0.5g / L, pH7.2, make up the insufficient part with pure water;

[0026] (3) Fermentation in fermenter: inoculate the seed solution in step (2) with 10% inoculu...

Embodiment 2

[0031] Strain activation and seed liquid preparation are the same as in Example 1

[0032] (3) Fermentation in fermenter: inoculate the seed solution in step (2) with 10% inoculum in the fermenter, the liquid volume of the fermentation medium in the 30L fermenter is 15L, the initial fermentation temperature is 37°C, and the tank pressure is 0.01Mpa for ventilation The volume is 0.5vvm, the speed is 400rpm; when the fermentation reaches 24 hours, adjust the speed to 200rpm, the ventilation volume is 1.0vvm, and start to add fed-batch medium, adjust the flow speed of fed-batch medium through the feedback of pH value, and control the pH value at 7.4 2 hours before the end of fermentation, the total fermentation time was 72h.

[0033] The fermentation medium consists of: glucose 80g / L, yeast extract 20g / L, NH4NO 3 4.5g / L, MgSO 4 ·6H 2 O0.5g / L, NaCl10g / L, CaCl 2 ·6H 2 O0.5g / L, FeSO 4 0.01g / L, sodium glutamate 40g / L, NaNO 3 20g / L,MnSO 4 ·H 2 O0.15g / L, pH value 7.4, insuffic...

Embodiment 3

[0037] (3) Fermentation in fermenter: inoculate the seed solution in step (2) with 10% inoculum in the fermenter, the liquid volume of the fermentation medium in the 30L fermenter is 15L, the initial fermentation temperature is 37°C, and the tank pressure is 0.05Mpa for ventilation The volume is 1.0vvm, the speed is 600rpm; when the fermentation reaches 24 hours, adjust the speed to 400rpm, the ventilation volume is 1.5vvm, and start to add fed-batch medium, adjust the flow speed of fed-batch medium through the feedback of pH value, and control the pH value at 7.9 6 hours before the end of fermentation, the total fermentation time was 72h.

[0038] The fermentation medium consists of: glucose 80g / L, yeast extract 20g / L, NH 4 NO 3 4.5g / L, MgSO 4 ·6H 2 O0.5g / L, NaCl10g / L, CaCl 2 ·6H 2 O0.5g / L, FeSO 4 0.01g / L, sodium glutamate 40g / L, NaNO 3 40g / L, MnSO 4 ·H 2 O0.15g / L, pH value 7.5, make up the insufficient part with pure water;

[0039] The fed-batch medium consists of...

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Abstract

The invention discloses a high-efficiency production method of gamma-polyglutamic acid, and belongs to the technical field of production of polymer materials based on biological fermentation. According to the method, bacillus licheniformis CGMCC (China General Microbiological Culture Collection Center) NO.3336 for high-efficient production of gamma-polyglutamic acid is taken as a starting strain, sodium nitrate is added into a fermentation culture medium, and the activity of polyglutamate synthase is influenced under the nitrate respiration condition by adjusting and controlling the pH value and the ventilation quantity in a fermentation process, so that residual glutamic acid in a fermentation solution is reduced and the conversion rate of glutamic acid is increased; after fermentation, the glutamic acid content of the fermentation solution is 5-15 g / L and the gamma-polyglutamic acid yield is 45-55 g / L. Therefore, the economic and feasible production method is provided for industrialization production of gamma-polyglutamic acid.

Description

technical field [0001] The invention belongs to the technical field of bio-fermentation production of polymer materials, in particular to a method for efficiently producing gamma-polyglutamic acid. Background technique [0002] γ-polyglutamic acid is a multifunctional biodegradable polymer material composed of D-glutamic acid and L-glutamic acid through the amide bond between α-amino group and γ-carboxyl group. Because γ-polyglutamic acid and its derivatives have excellent water solubility and adsorption properties, can be completely biodegraded, are safe and non-toxic to humans and animals, and can be eaten, they can be used as water-retaining agents, thickeners, flocculation agents, etc. Agents, heavy metal adsorbents, drugs, fertilizer slow-release agents and drug carriers, etc., have broad application prospects in the fields of agriculture, food, medicine, cosmetics, environmental protection, synthetic fibers and coatings. With the increasingly serious environmental pol...

Claims

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Application Information

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IPC IPC(8): C12P13/00C12R1/10
Inventor 乔长晟高明昊李小鑫张帅李雪
Owner 天津北洋百川生物技术有限公司
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