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Process for producing gamma-polyglutamic acid

A technology of polyglutamic acid and products, which is applied in the field of preparation of γ-polyglutamic acid, can solve the problems of relatively high cost of raw materials and low yield, and achieve the effects of cost advantage, high yield and low cost

Inactive Publication Date: 2009-04-08
GUANGDONG INST OF MICROORGANISM
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

According to whether L-glutamic acid is needed for the nutritional requirements of cell growth, γ-PGA-producing bacteria can be divided into two categories. One type needs L-glutamic acid to accumulate γ-PGA during cultivation. Such bacteria mainly include B. anthracis, B. subtilis MR-141, B. licheniformis ATCC 9945, B. licheniformis ATCC 9945a, B. subtilis IFO3335 and B. subtilis F-2-01, etc., the γ-PGA production of these strains is relatively high, generally 15-35g / L, but the relative cost of raw materials is relatively high; one type can accumulate γ-PGA without L-glutamic acid during cultivation, such as B.subtilis 5E, B.subtilis var.polyglutamicum, B.licheniformisA35 , B.subtilis TAM-4, etc., γ-PGA producing bacteria that do not require glutamic acid, the γ-PGA yield is low, generally 5-10g / L, but because glutamic acid can be replaced by cheap raw materials, it is widely used in industry It has practical application significance in production

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Take the slant species preserved in the refrigerator at 4°C for 1 day [composition of the preserved slant medium (g / L): peptone 8, NaCl 11, yeast extract 3, agar 20, pH7.0], and inoculate the above slant with an inoculation needle for about 1cm 2 Scrape off the strains of the area and transfer to the newly configured seed liquid activation medium [liquid activation medium composition (g / L): citric acid 15, NH 4 Cl 8, peptone 7, yeast powder 15, corn steep liquor 15, NaCl 3, dissolved in tap water and then adjusted to 1 L, adjusted to pH 6.5], the liquid activation medium was packed in a 250 mL Erlenmeyer flask, and the amount of the Erlenmeyer flask was 25 mL. Shake culture in a reciprocating shaker at 32°C for 18 hours, with a shaker rotation speed of 110 r / m and a stroke of 75 mm. Then transfer in the fermentation medium by 5% inoculum size, the composition of fermentation medium is as follows: glucose 40g, NH 4 Dissolve 12g of Cl, 14g of peptone, 38g of yeast powde...

Embodiment 2

[0025]Take the slant species preserved in the refrigerator at 4°C for 3 days [the composition of the preserved slant medium (g / L): peptone 12, NaCl 9, yeast extract 8, agar 16, pH6.8], and inoculate the above slant with an inoculation needle for about 1cm 2 Scrape off the bacteria in the area and transfer to the newly configured seed liquid activation medium [liquid activation medium composition (g / L): citric acid 18, NH 4 Cl 8, peptone 5, yeast powder 20, corn steep liquor 10, NaCl 4.5, dissolved in tap water and adjusted to 1 L, adjusted to pH 6.0], the liquid activation medium was packed in a 250 mL Erlenmeyer flask, and the volume of the Erlenmeyer flask was 25 mL. Shake culture in a reciprocating shaker at 34°C for 22 hours, the shaker speed is 80r / m, and the stroke is 65mm. Then transfer in the fermentation medium by 5% inoculum size, the composition of fermentation medium is as follows: glycerol 35g, fructose 2g, NH 4 Dissolve 20g of Cl, 10g of peptone, 45g of yeast p...

Embodiment 3

[0027] Take the slant species preserved in the refrigerator at 4°C for 3 days [the composition of the preserved slant medium (g / L): peptone 10, NaCl 10, yeast extract 5, agar 18, pH 7.2], and use an inoculation needle to inoculate the above slant for about 1cm 2 The strains of the area were scraped off and transferred to the newly configured seed liquid activation medium [liquid activation medium composition (g / L): citric acid 20, (NH 4 ) 2 SO 4 8. Peptone 6, yeast powder 17, corn steep liquor 13, NaCl 5, dissolved in tap water to 1L, adjusted to pH 7.0], liquid activation medium was packed in a 250mL triangular flask with a capacity of 25mL, at 37 Cultivate in a reciprocating shaker for 24 hours with shaking at a speed of 95 r / m and a stroke of 70 mm. Then transfer in the fermentation medium by 5% inoculum size, the composition of fermentation medium is as follows: 30g of citric acid, 4g of fructose, NH 4 Dissolve 16g of Cl, 12g of peptone, 30g of yeast powder, 20g of cor...

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Abstract

The invention provides a preparation method of gamma-polyglutamic acid, the method uses Bacillus subtilis PGA-7 CCTCC M 206102 as a fermentation strain to carry out the flask-shaking fermentation of a fermentation culture medium so as to prepare the gamma-polyglutamic acid, and the process comprises the slant strain activation, the liquid activation of the strain, the flask-shaking fermentation and the flask-shaking fermented mash extraction. The preparation method adopts the preferential fermentation raw materials combined by a variety of organic nitrogen sources and inorganic nitrogen sources, the highest gamma-PGA production can be 27.3g / l under the best flask-shaking fermentation process conditions, thereby having good industrial application prospect.

Description

technical field [0001] The invention relates to a method for preparing gamma-polyglutamic acid, in particular to a method for preparing gamma-polyglutamic acid by using Bacillus subtilis PGA-7CCTCC M 206102 for shake flask fermentation. Background technique [0002] γ-polyglutamic acid (γ-PGA) is an amino acid polymer that can be biosynthesized by microorganisms in large quantities. It is composed of D-type or L-type glutamic acid linked by γ-amide bonds. γ-PGA synthesized by microorganisms is a water-soluble biodegradable biopolymer, usually composed of about 5,000 glutamic acid monomers, and the relative molecular mass is generally 100,000 to 1 million. γ-PGA can be made into different molecular weights and used in various fields. For example: (1) γ-PGA is food safe and can be used as dietary fiber, health food, food thickener, stabilizer or as a moisturizer for cosmetics; (2) γ-PGA can be made into water gel, which has extremely high Water absorption capacity, can absor...

Claims

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Application Information

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IPC IPC(8): C12P13/00C12R1/125
Inventor 疏秀林施庆珊冯静李诚斌欧阳友生谢小保陈仪本
Owner GUANGDONG INST OF MICROORGANISM
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