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Non-dependent production method of gamma-polyglutamic acid from glutamic acid

A technology of polyglutamic acid and production method, applied in the field of biological sciences, can solve the problems of high raw material cost, unsuitable for industrial scale production, low production efficiency, etc., and achieve the effects of increased output, easy operation, and short cycle

Inactive Publication Date: 2012-07-18
ZHEJIANG DEQING HUINING BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the current method of fermentative production of polyglutamic acid, glutamic acid needs to be added as a precursor in the medium, resulting in high raw material costs and becoming one of the main bottlenecks in industrial production and application.
The fermentation process without adding glutamic acid can only produce low-concentration polyglutamic acid, and the production efficiency is too low to be suitable for industrial scale production

Method used

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  • Non-dependent production method of gamma-polyglutamic acid from glutamic acid
  • Non-dependent production method of gamma-polyglutamic acid from glutamic acid
  • Non-dependent production method of gamma-polyglutamic acid from glutamic acid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] The glutamate-independent production method of γ-polyglutamic acid comprises the following steps: step 1) strain activation: streak Bacillus subtilis C10 on a plate, culture at 37° C. for 12 hours. The plate medium consists of: peptone 10g / L, beef extract 5g / L, sodium chloride 5g / L, glucose 10g / L, agar 15g / L, pH 7.

[0021] Step 2) Preparation of seeds: pick the activated Bacillus subtilis (Bacillus subtilis) C10 bacterial classification and insert it into the liquid seed culture medium, the liquid filling volume is 20ml culture medium in a 250ml Erlenmeyer flask, 37°C, 200rpm (rev / min), Shake culture for 12h. The composition of the above seed medium: peptone 10g / L, beef extract 5g / L, sodium chloride 5g / L, glucose 10g / L, pH 7.

[0022] Step 3) Put the prepared seed liquid into a 10L fermenter at a ratio of 5%, the liquid content of the fermentation medium is 6L, and the fermentation parameters are controlled as follows: fermentation temperature 37°C, stirring 400rpm, p...

Embodiment 2

[0026] Step 1) is the same as step 1) in Example 1.

[0027] Step 2) is the same as step 2) in Example 1.

[0028] Step 3) and step 3 in embodiment 1) except following difference, all the other technical features are identical, and difference is that when citric acid concentration is lower than 5g / L in the fermentation medium, start to add the mixing of citric acid and ammonium chloride intermittently The concentration of the solution is 250g / L and 48g / l respectively, the concentration of citric acid in the maintenance medium is about 5-20g / L, and the concentration of ammonium chloride is 1-10g / L. Stop feeding when the concentration of γ-PGA in the medium no longer increases.

[0029] Step 4) is the same as step 4) in Example 1.

[0030] The fermentation results show that the final consumption of glucose is 57g / L, the consumption of citric acid is 29.8g / L, and the output of γ-PGA is 38.2g / L. The fermentation process curve is shown in image 3 .

Embodiment 3

[0041] Step 1) is the same as step 1) in Example 1.

[0042] Step 2) is the same as step 2) in Example 1.

[0043] Step 3) Put the prepared seed liquid into a 10L fermenter at a ratio of 5%, the liquid content of the fermentation medium is 6L, and the fermentation parameters are controlled as follows: fermentation temperature 37°C, stirring 400rpm, pH 6.5, ventilation 0.4L / h. When the concentration of succinic acid in the fermentation medium is lower than 5g / L, start intermittently feeding sodium succinate solution with a concentration of 50g / L to maintain the concentration of succinic acid in the medium at about 5-10g / L. Stop feeding when the concentration of γ-PGA in the medium no longer increases.

[0044] The composition of the above fermentation medium: glucose 80g / l, ammonium chloride 10g / l, succinic acid 20g / l, KH2PO4 1.5g / l, MgSO4 7H2O 0.5g / l, FeCl3 0.05g / l, MnSO4 H2O 0.1g / l, CaCl2 0.11g / l, pH 6.5.

[0045] Step 4) is the same as step 4) in Example 1.

[0046] F...

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Abstract

The invention discloses and provides a non-dependent production method of gamma-polyglutamic acid from glutamic acid. The gamma-polyglutamic acid produced by the prior art is low in concentration and high in cost. According to the non-dependent production method disclosed by the invention, an organic acid or a mixture of the organic acid and a nitrogen source is added into a culture medium by flowing during the process of culturing aerobic bacillus sp. to induce thalli to transform a saccharine material and the organic acid into the gamma-polyglutamic acid, and a precursor L-glutamic acid does not need to be additionally added into the culture medium. The fermentation method for producing the gamma-polyglutamic acid, disclosed by the invention, is short in period and easy to operate, can be used for industrial large-scale fermentation and has great application value.

Description

technical field [0001] The invention belongs to the technical field of biological sciences, and in particular relates to a new method for glutamate-independent production of gamma-polyglutamic acid. Background technique [0002] γ-polyglutamic acid (Poly-γ-glutamatic acid, γ-PGA) is a water-soluble and biodegradable polyamino acid produced by a variety of microorganisms. Polyglutamic acid is a new type of natural polymer that is non-toxic to the human body and the environment. Its unique structural properties endow it with high water absorption, good permeability, biocompatibility and biodegradability. It is widely used in agriculture, medicine It is recognized as a green chemical product with great development potential in many fields such as food, cosmetics, etc. The production of γ-polyglutamic acid mainly uses some strains of Bacillus genus for direct fermentation. Compared with polymers based on petroleum derivatives, γ-polyglutamate can be synthesized from renewable ...

Claims

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Application Information

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IPC IPC(8): C12P13/02C12R1/125C12R1/10C12R1/11C12R1/07
Inventor 徐志南张慧莉黄磊蔡谨
Owner ZHEJIANG DEQING HUINING BIOTECH
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