Method for increasing dissolved oxygen in gamma-polyglutamic acid fermenting liquid

A technology of polyglutamic acid and fermentation liquid, applied in the field of microbial fermentation, can solve the problems of low salinity tolerance, lack of applicability, limitations, etc., and achieve low cost, good application prospects, and no tolerance requirements Effect

Active Publication Date: 2017-07-14
CHENGDU INST OF BIOLOGY CHINESE ACAD OF S
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Problems solved by technology

However, this patented method is only suitable for high-salt-tolerant strains, and is not applicable to most strains with low salinity tolerance
Li Haijun et al. (publication number CN 103881954 A) recombined and integrated the hemoglobin gene (vgb) of Vitiligo hyaline on the chromosome of Bacillus subtilis strain FRD518, and successfully expressed the hemoglobin VHb of Vitiligo hyaline at high levels, which significantly improved the low-dissolved oxygen conditions of recombinant Bacillus subtilis. Under the utilization rate of oxygen, the yield of gamma-polyglutamic acid of the recombinant strain reached 65g / L, which was 147% higher than that of the original strain, but factors such as the genetic instability of the genetic engineering strain and the safety issues of the genetic engineering strain also limited the Application of engineering strains in fermentative preparation of γ-polyglutamic acid

Method used

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  • Method for increasing dissolved oxygen in gamma-polyglutamic acid fermenting liquid
  • Method for increasing dissolved oxygen in gamma-polyglutamic acid fermenting liquid
  • Method for increasing dissolved oxygen in gamma-polyglutamic acid fermenting liquid

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Experimental program
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Effect test

Embodiment 1

[0030] 1. Prepare medium:

[0031] a. Activation medium components: beef extract 5g, peptone 10g, NaCl 5g, agar 20g, water up to 1000mL, pH 7.0-7.4, sterilized at 121°C for 30min.

[0032] b. Seed medium components: beef extract 5g, peptone 10g, NaCl 5g, water up to 1000mL, pH 7.0-7.4, sterilized at 121°C for 30min.

[0033] c. Fermentation medium components: citric acid 10g, glutamic acid 20g, ammonium sulfate 10g, glycerin 80g, K 2 HPO 4 1g, MgSO 4 ·7H 2 O 0.5g, FeCl 3 ·6H 2 O 0.02g, MnSO 4 ·H 2 O 0.1g, CaCl 2 0.2g, pH 7.0, add water to 1000mL, sterilize at 121°C for 30min.

[0034] 2. Activate the strain and prepare the seed solution:

[0035] The cryopreserved Bacillus amyloliquefaciens JX-6 was inoculated on the slant activation medium for activation, and cultured at 37°C for 24h. The activated Bacillus amyloliquefaciens JX-6 was inoculated into the seed medium, and cultured on a shaker at 37° C. with a rotation speed of 150 r / min for 24 hours to obtain the s...

Embodiment 2

[0052] 1. Prepare medium:

[0053] a. Activation medium components: beef extract 5g, peptone 10g, NaCl 5g, agar 20g, water up to 1000mL, pH 7.0-7.4, sterilized at 121°C for 30min.

[0054] b. Seed medium components: beef extract 5g, peptone 10g, NaCl 5g, water up to 1000mL, pH 7.0-7.4, sterilized at 121°C for 30min.

[0055] c. Fermentation medium components: citric acid 10g, glutamic acid 20g, ammonium sulfate 10g, glycerin 80g, K 2 HPO 4 1g, MgSO 4 ·7H 2 O 0.5g, FeCl 3 ·6H 2 O 0.02g, MnSO 4 ·H 2 O 0.1g, CaCl 2 0.2g, pH 7.0, add water to 1000mL, sterilize at 121°C for 30min.

[0056] 2. Activate the strain and prepare the seed solution:

[0057] The cryopreserved Bacillus amyloliquefaciens JX-6 was inoculated on the slant activation medium for activation, and cultured at 35°C for 36h. The activated Bacillus amyloliquefaciens JX-6 was inoculated into the seed medium, and cultured on a shaker at 35° C. with a rotation speed of 200 r / min for 36 hours to obtain the s...

Embodiment 3

[0071] 1. Prepare medium:

[0072] a. Activation medium components: beef extract 5g, peptone 10g, NaCl 5g, agar 20g, water up to 1000mL, pH 7.0-7.4, sterilized at 121°C for 30min.

[0073] b. Seed medium components: beef extract 5g, peptone 10g, NaCl 5g, water up to 1000mL, pH 7.0-7.4, sterilized at 121°C for 30min.

[0074] c. Fermentation medium components: citric acid 50g, glutamic acid 100g, ammonium sulfate 50g, glycerin 400g, K 2 HPO 4 5g, MgSO 4 ·7H 2 O 2.5g, FeCl 3 ·6H 2 O 0.1g, MnSO 4 ·H 2 O 0.5g, CaCl 2 1g, pH 7.0, add water to 5L, sterilize at 121°C for 30min.

[0075] 2. Activate the strain and prepare the seed solution:

[0076] The cryopreserved Bacillus amyloliquefaciens JX-6 was inoculated on the slant activation medium for activation, and cultured at 37°C for 24h. The activated Bacillus amyloliquefaciens JX-6 was inoculated into the seed medium, and cultured on a shaker at 37° C. with a rotation speed of 150 r / min for 24 hours to obtain the seed c...

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Abstract

The invention belongs to the field of microorganism fermenting, which aims at providing a method for increasing dissolved oxygen in gamma-polyglutamic acid fermenting liquid with universality. The method adopts the technical scheme that the collection number of bacillus amyloliquefaciens JX-6 is CGMCC No.13715, and comprises the following steps of inoculating the bacteria strain into a sterilized oblique activation culture medium, and culturing, so as to obtain activated bacterial strain; inoculating the activated bacteria strain into a sterilized and cooled seed culture medium, and culturing, so as to obtain a seed liquid; inoculating the seed liquid into a fermenting culture medium, adding solid oxygen in the fermenting process, and culturing, so as to obtain the fermenting liquid. The method has the advantages that the universality is strong, the operation is simple, the cost is low, the secondary pollution is avoided, the resistance requirement on the bacteria strain is avoided, the method is suitable for large-scale production, and the application prospect is good.

Description

technical field [0001] The invention belongs to the field of microbial fermentation, and in particular relates to a method for increasing dissolved oxygen in gamma-polyglutamic acid fermentation liquid. Background technique [0002] γ-polyglutamic acid is a water-soluble biopolymer formed by combining L-glutamic acid or D-glutamic acid through γ-amide groups. It has strong water absorption, degradability, hydrolysis and many other characteristics, so it is widely used in many fields such as food, agriculture, medicine, greening and water treatment, and has great development value and application prospect. [0003] The preparation methods of γ-polyglutamic acid include chemical synthesis, biological extraction and microbial fermentation. The production cost of microbial fermentation method is lower than the previous two methods, and the production process has little pollution to the environment, so now the microbial fermentation method is mainly used to produce γ-polyglutami...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P13/02C12R1/07
CPCC12P13/02C12N1/205C12R2001/07
Inventor 闫志英姬高升许力山房俊楠刘晓风
Owner CHENGDU INST OF BIOLOGY CHINESE ACAD OF S
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