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A kind of production method of high molecular weight gamma-polyglutamic acid

A technology of polyglutamic acid and production method, which is applied in the field of biological fermentation to produce polymer materials, can solve the problems of unclear molecular weight range of γ-polyglutamic acid, difficult control of the molecular weight range of γ-polyglutamic acid, etc. High, simple process effect

Active Publication Date: 2015-11-18
四川百川金开生物工程有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] The molecular weight range of γ-polyglutamic acid is difficult to control, and the specific molecular weight range of the obtained γ-polyglutamic acid is not yet clear. Therefore, it is necessary to explore a large-scale production of stable high molecular weight γ-polyglutamic acid The method is imminent. At present, there is no literature report on the production of high molecular weight γ-polyglutamic acid by fermentation of Bacillus licheniformis

Method used

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  • A kind of production method of high molecular weight gamma-polyglutamic acid
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  • A kind of production method of high molecular weight gamma-polyglutamic acid

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Experimental program
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Effect test

Embodiment 1

[0026] A production method of high molecular weight gamma-polyglutamic acid, comprising the steps of:

[0027] (1) Strain activation: the original strain of Bacillus licheniformis CGMCCNO.3336 was inoculated on the slant medium, cultured at 37°C for 16 hours, activated 2-3 times in this way, and mature slant seeds were prepared;

[0028] 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;

[0029] (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;

[0030] 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;

[0031](3) Fermentation in fermenter: inoculate the seed solution in step (2) with 1...

Embodiment 2

[0036] Strain activation and seed liquid preparation are the same as in Example 1

[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 39°C, and the tank pressure is 0.03Mpa for ventilation Volume 0.7vvm, speed 500rpm; when the fermentation broth OD 660 At 0.570, control the fermentation temperature to 32°C, adjust the rotation speed to 300rpm, and the ventilation rate to 1.8vvm, and start to add feed medium, control the pH at 7.4 to 4 hours before the end of fermentation, and the fermentation time is 72h.

[0038] The fermentation medium consists of: glucose 50g / L, yeast extract 20g / L, NH 4 NO 3 4.5g / L, MgSO 4 ·6H 2 O1.0g / L, NaCl10g / L, CaCl 2 ·6H 2 O1.2g / L, FeSO 4 0.009g / L, sodium glutamate 75g / L, pH value 7.4, make up the deficiency with purified water;

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

Embodiment 3

[0042] (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 40°C, and the tank pressure is 0.05Mpa for ventilation Volume 1.0vvm, speed 600rpm; when the fermentation broth OD 660 At 0.600, control the fermentation temperature to 35°C, adjust the rotation speed to 350rpm, and the ventilation rate to 2.0vvm, and start to add feeding medium, control the pH at 7.5 to 6 hours before the end of fermentation, and the fermentation time is 72h.

[0043] The fermentation medium consists of: glucose 60g / L, yeast extract 25g / L, NH 4 NO 3 5.5g / L, MgSO 4 ·6H 2 O1.2g / L, NaCl12g / L, CaCl 2 ·6H 2 O1.5g / L, FeSO 4 0.010g / L, sodium glutamate 80g / L, pH value 7.5, make up the deficiency with purified water;

[0044] The fed-batch medium consists of: glucose 850g / L, sodium chloride 60.0g / L, CaCl 2 ·6H 2 O15.0g / L, MnSO 4 10g / L...

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Abstract

The invention discloses a production method of a gamma-polyglutamic acid with high molecular weight, and belongs to the technical field of biological fermentation production of polymer materials. Bacillus licheniformis CGMCC NO.3336 is taken as an original strain, the rotating speed is reduced at a later stage of fermentation, the effect on polyglutamic acid synthesis caused by shearing force is reduced by regulating and controlling of ventilation and rotation speed in the fermentation process, and the temperature is changed, so as to affect the activity of folylpolyglutamate synthetase. Especially, the fermentation pH is controlled and the ion strength in the culture medium is increased by feeding glucose, sodium chloride, CaCl2.6H2O, MnSO4 and MgSO4, so as to obtain the polyglutamic acid with accurate molecular weight. The molecular weight is high, and the molecular weight of the gamma-polyglutamic acid is 1000000-4000000 Dalton by detection after fermentation is finished. The method is simple in technology, and the gamma-polyglutamic acid is high in molecular weight and relatively stable and controllable, and large-scale production can be achieved.

Description

Technical field: [0001] The invention belongs to the technical field of biofermentation production of polymer materials, and in particular relates to a production method of high molecular weight 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, drug / 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 serio...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P13/02C12R1/10
Inventor 乔长晟高明昊李小鑫张帅李雪
Owner 四川百川金开生物工程有限公司
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