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Bacillus subtilis for producing gamma-polyglutamic acid by high-temperature fermentation and application of bacillus subtilis for producing gamma-polyglutamic acid by high-temperature fermentation

A Bacillus subtilis, high-temperature fermentation technology, applied in the field of microorganisms, can solve problems such as no published literature reports, and achieve the effects of simple and easy cultivation methods, simple nutritional requirements, and excellent temperature characteristics.

Active Publication Date: 2014-07-30
洛阳丰侨生物技术研究所有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Bacillus subtilis currently used to produce γ-polyglutamic acid, most of its isolated strains come from soil, fermented bean curd, pickled radish sticks, pickled cucumber, pickled garlic, soybean paste, watermelon sauce, pickles, sausage, soybean drum , natto, and bacon. There is no public literature report on the use of sea sand as the material to isolate strains

Method used

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  • Bacillus subtilis for producing gamma-polyglutamic acid by high-temperature fermentation and application of bacillus subtilis for producing gamma-polyglutamic acid by high-temperature fermentation
  • Bacillus subtilis for producing gamma-polyglutamic acid by high-temperature fermentation and application of bacillus subtilis for producing gamma-polyglutamic acid by high-temperature fermentation
  • Bacillus subtilis for producing gamma-polyglutamic acid by high-temperature fermentation and application of bacillus subtilis for producing gamma-polyglutamic acid by high-temperature fermentation

Examples

Experimental program
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Effect test

Embodiment 1

[0045] 1. Separation of CCTCC NO: M2012347

[0046] Preparation of bacterial suspension: Weigh 1g of sea sand sample into a 250ml Erlenmeyer flask filled with 100ml of sterile saline, stir magnetically at room temperature for 30 minutes to prepare bacterial suspension, and treat at 100°C for 10min.

[0047] Bacterial suspension dilution and colony picking: The bacterial suspension obtained above was diluted into bacterial suspensions of different concentrations using the 10-fold dilution method, and 10 5 , 10 6 , 10 7 100 μl of the bacterial suspension was spread on the solid separation medium plate; after 24 hours of constant temperature incubation at 50°C, pick a single colony with a viscous colony surface that can be picked up with a toothpick.

[0048] Streak purification: use a flat and smooth inoculation loop to continuously streak and purify the obtained single colony on a solid separation medium plate under aseptic conditions, and pick the pure species on the plate a...

Embodiment 2

[0091] 1. Separation of CCTCC NO: M2012347

[0092] Preparation of bacterial suspension: Weigh 1g of sea sand sample into a 250ml Erlenmeyer flask filled with 80ml of sterile saline, stir magnetically at room temperature for 25 minutes to prepare bacterial suspension, and treat at 90°C for 15min.

[0093] Bacterial suspension dilution and colony picking: The bacterial suspension obtained above was diluted into bacterial suspensions of different concentrations using the 10-fold dilution method, and 10 5 , 10 6 , 10 7 100 μl of the bacterial suspension was spread on the solid separation medium plate; after 20 hours of constant temperature incubation at 45°C, pick a single colony with a viscous colony surface that can be picked up with a toothpick.

[0094] Streak purification: use a flat and smooth inoculation loop to continuously streak and purify the obtained single colony on a solid separation medium plate under aseptic conditions, and pick the pure species on the plate aft...

Embodiment 3

[0100] 1. Separation of CCTCC NO: M2012347

[0101] Preparation of bacterial suspension: Weigh 1g of sea sand sample into a 250ml Erlenmeyer flask filled with 120ml of sterile saline, stir magnetically at room temperature for 40 minutes to prepare bacterial suspension, and treat at 110°C for 8 minutes.

[0102] Bacterial suspension dilution and colony picking: The bacterial suspension obtained above was diluted into bacterial suspensions of different concentrations using the 10-fold dilution method, and 10 5 , 10 6 , 10 7 100 μl of the bacterial suspension was spread on the solid separation medium plate; after 25 hours of constant temperature incubation at 60°C, pick a single colony with a viscous surface and a toothpick that can be picked up and drawn.

[0103] Streak purification: use a flat and smooth inoculation loop to continuously streak and purify the obtained single colony on a solid separation medium plate under aseptic conditions, and pick the pure species on the p...

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Abstract

The invention discloses a bacillus subtilis for producing gamma-polyglutamic acid by high-temperature fermentation, wherein the classification name is Bacillus subtilis GXA-28; the preservation number is CCTCCNO: M2012347; the preservation data is 14th, September, 2012; and the preservation unit is China Center for Type Culture Collection. The Bacillus subtilis GXA-28 is applied in producing gamma-polyglutamic acid by high-temeprature fermentation at 40-50 DEG C. The bacillus subtilis is separated from sea sands, and can effectively synthesize gamma-polyglutamic acid at high temperature; the highest temperature is 50 DEG C; the fermentation time is 18-25h; the yield can reach 20-30g / l; the production rate can arrive at 0.9-1.3g / l.h; and the bacillus subtilis has the advantages of excellent temperature characteristic, simple nutritional requirement and simple culture method, which can provide good conditions for industrial production.

Description

technical field [0001] The invention belongs to the field of microorganisms, and in particular relates to a bacillus subtilis used for high-temperature fermentation to produce gamma-polyglutamic acid and its application. Background technique [0002] γ-polyglutamic acid (Poly(γ-glutamic acid), γ-PGA) is a unique biologically high molecular. It is usually polymerized by 500-5000 glutamic acid monomers, and its molecular weight is between 100-1000kDa. The glutamic acid residues in the γ-polyglutamic acid molecule have a large number of free hydrophilic carboxyl groups, which can form hydrogen bonds inside or between molecules, and have extremely high water solubility and water absorption and moisture retention; these free carboxyl groups At the same time, it provides a cation-bonding group, which makes it have good adsorption to metal ions. In addition, as an environmentally friendly biopolymer, γ-polyglutamic acid also has some unique and excellent properties, such as edib...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12P13/02C12R1/125
Inventor 梁智群曾伟陈桂光张云开
Owner 洛阳丰侨生物技术研究所有限公司