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Fermentation method for efficient coproduction of poly-gamma-glutamic acid and 2,3-butanediol by virtue of bacillus

The technology of Bacillus licheniformis and Bacillus licheniformis, which is applied in the field of microbial fermentation, can solve the problems of limiting the application of production strains, low utilization rate of raw materials, and high cost, and achieves significant co-production yield, low raw material cost, and increased co-production yield. Effect

Inactive Publication Date: 2015-04-29
武汉康绿达生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the theoretical yield of 2,3-butanediol to glucose is only about 0.5 g / g, and most of the current high-yielding 2,3-butanediol strains are pathogenic bacteria, such as Serratia marcescens and Enterobacter aerogenes , Klebsiella, etc., and the safety principles of large-scale industrial production limit the application of these production strains
[0004] After retrieval, the existing reported technologies only produce poly-γ-glutamic acid or 2,3-butanediol alone, which has problems such as low raw material utilization and high cost.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Embodiment 1 uses bacillus licheniformis to co-produce poly-γ-glutamic acid and 2,3-butanediol with fermentation medium 1 shake flask fermentation

[0020] (1) Use the frozen-preserved strain Bacillus licheniformis WX-02 (CCTCC NO: M208065) ​​as the production strain.

[0021] (2) Activation of slant strains:

[0022] The cryopreserved Bacillus licheniformis WX-02 was inoculated on LB solid slant (containing peptone 10g / L, yeast extract 5g / L, NaCl 10g / L, agar 15g / L, pH7.0), cultured at 37°C for 24h to obtain Activate the bacteria.

[0023] (3) Seed liquid culture:

[0024] Transfer the seed bacteria activated on the LB solid slant to the LB triangular flask liquid seed medium containing peptone 10g / L, yeast extract 5g / L, NaCl 10g / L, pH 7.0, and cultivate at 37°C and 200r / min 10h to mid-logarithmic growth phase. (4) Shake flask batch fermentation culture:

[0025] The preparation method of the above-mentioned fermentation medium 1 is as follows: glucose 90g / L, sodiu...

Embodiment 2

[0031] Embodiment 2 uses bacillus licheniformis to co-produce poly-γ-glutamic acid and 2,3-butanediol with fermentation medium 2 shake flask fermentation

[0032] (1) The production strain used in this example is the same as that in Example 1 (1).

[0033] (2) The activation of strains on the slant is the same as in Example 1 (2).

[0034] (3) Seed liquid cultivation is the same as in Example 1 (3).

[0035] (4) Shake flask batch fermentation culture:

[0036] The preparation method of the above-mentioned fermentation medium 2 is as follows: glucose 90g / L, sodium glutamate 30g / L, sodium nitrate 10g / L, sodium citrate 10g / L, ammonium chloride 8g / L, MnSO 4 ·H 2 O0.15g / L, MgSO 4 1g / L, K 2 HPO 4 ·3H 2O1g / L, ZnSO 4 ·7H 2 O1g / L, CaCl 2 1g / L, pH 7.0, add distilled water to 1L, divide the fermentation medium into 500mL Erlenmeyer flasks, the volume of each bottle is 100mL, sterilize at 115°C for 30min, cool to 37°C, and inoculate according to 1.5ml The seed solution was inse...

Embodiment 3

[0040] Example 3 Co-production of poly-γ-glutamic acid and 2,3-butanediol with fermentation medium 3 shake flasks by Bacillus licheniformis

[0041] (1) The production strain used in this example is the same as that in Example 1 (1).

[0042] (2) The activation of strains on the slant is the same as in Example 1 (2).

[0043] (3) Seed liquid cultivation is the same as in Example 1 (3).

[0044] (4) Shake flask batch fermentation culture:

[0045] The preparation method of the above-mentioned fermentation medium 3 is as follows: glucose 90g / L, sodium glutamate 80g / L, sodium nitrate 10g / L, sodium citrate 10g / L, ammonium chloride 8g / L, MnSO 4 ·H 2 O0.15g / L, MgSO 4 0.5g / L, K 2 HPO 4 ·3H 2 O0.5g / L, ZnSO 4 ·7H 2 O0.5g / L, CaCl 2 0.5g / L, pH7.0, add distilled water to 1L, divide the fermentation medium into 500mL Erlenmeyer flasks, each bottle contains 100mL, sterilize at 115°C for 30min, cool to 37°C, and inoculate according to 1.5ml The amount of seed solution was added, t...

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Abstract

The invention belongs to the technical field of microbial fermentation and particularly relates to a batch fermentation method for efficient coproduction of poly-gamma-glutamic acid and 2,3-butanediol by virtue of bacillus. According to the method, by liquid-submerged fermentation, two products, namely, poly-gamma-glutamic acid and 2,3-butanediol are simultaneously produced. The process can be used for safely producing the strain, the production cost is effectively reduced, the highest concentrations of simultaneously produced poly-gamma-glutamic acid and 2,3-butanediol can respectively reach 52.2g / L and 73.68g / L, the apparent conversion rate of glucose reaches 83.9%, the production strength reaches 2.10g.L<-1>.h<-1>, and the method is suitable for industrial production.

Description

technical field [0001] The invention belongs to the technical field of microbial fermentation, and in particular relates to a method for efficiently co-producing poly-γ-glutamic acid and 2,3-butanediol by batch fermentation with bacillus. Background technique [0002] Poly-γ-glutamic acid (γ-PGA) is a homopolyamide formed by linking L- and D-glutamic acid. It is a high-molecular extracellular polymer that can be synthesized by microorganisms. The average relative molecular weight is generally between 10-10000 kD. γ-PGA is water-soluble, highly viscous, film-forming and moisturizing, and is non-toxic to humans and the environment. Therefore, it has broad application prospects in the fields of pharmaceutical manufacturing, environmental governance, light industry and agriculture. Because the currently screened microorganisms have various problems such as low yield of γ-PGA and high cost of raw materials, the industrial application is severely restricted. How to reduce the f...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P13/02C12P7/18C12R1/10C12R1/125
Inventor 陈守文高立冀志霞魏雪团祁高富
Owner 武汉康绿达生物科技有限公司
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