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A strain of Bacillus subtilis with high yield of γ-pga and its application

A technology of Bacillus subtilis, CGMCCNO.17215, applied in the field of microorganisms, can solve the problems of incapable of large-scale production, long fermentation cycle, high energy consumption of fermentation, etc., and achieve the effects of increasing yield, improving permeability, and low energy consumption

Active Publication Date: 2022-04-26
CHENGDU INST OF BIOLOGY CHINESE ACAD OF S
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, the existing γ-PGA is generally obtained by microbial fermentation, which has defects such as long fermentation period, high fermentation energy consumption, and low efficiency, and cannot be mass-produced

Method used

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  • A strain of Bacillus subtilis with high yield of γ-pga and its application
  • A strain of Bacillus subtilis with high yield of γ-pga and its application
  • A strain of Bacillus subtilis with high yield of γ-pga and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Embodiment one: the acquisition and identification of Bacillus subtilis

[0040] 1. Obtaining the strain

[0041](1) Extract landfill leachate from a waste treatment plant in Chengdu, add 10g of leachate to 90mL sterile water to get 10 -1 Dilution of diluent. Incubate the diluted solution in a water bath at 80°C for 15 minutes to kill most of the bacteria, then take 1mL of 10 -1 Dilute solution in 9mL sterile water to get 10 -2 Dilution of diluent. Dilute sequentially as above to obtain 10 -3 、10 -4 、10 -5 、10 -6 、10 -7 、10 -8 If the diluent is diluted, the subsequent dilution can be carried out directly, without the need for water bath insulation. Draw 0.2mL of the dilutions of each dilution and apply them on the corresponding E-type separation medium plate, and make corresponding labels (media name, sample name, dilution factor, incubation time), and then place them in the incubator. Culture upside down at 35°C (cultivate at 25-37°C).

[0042] (2) After th...

Embodiment 2

[0054] Example 2: The optimum fermentation temperature display of strain 8-2

[0055] 1. The strain 8-2 was inoculated on a slant medium for activation, and cultured at 35° C. for 12 hours. Inoculate the strain activated on the slant into the seed medium, and culture it at 35° C. for 12 hours at a shaker rotation speed of 180 rpm to obtain activated seed liquid. The seed liquid of described activation is inserted in the shake flask fermentation medium, the inoculum size is 5% (v / v), the liquid volume of the shake flask is 60mL / 500mL, the shaking table speed is 220rpm, and different temperatures (25° C., 28° C. , 30°C, 32°C, 35°C, 37°C, 40°C) for 24 hours.

[0056] After the fermentation is over, take the fermentation broth, add 6mol / L hydrochloric acid to adjust the pH value below 3, filter and remove the bacteria, then add 6mol / L sodium hydroxide to adjust the pH to neutral, add 3 times the volume of absolute ethanol, and centrifuge Purification of γ-PGA. Then at 110° C. i...

Embodiment 3

[0057] Example 3: Demonstration of the effect of plant growth promotion after γ-PGA activated biochar

[0058] The soil samples for the test were taken from farmland soil in Shuangliu County, Chengdu City, Sichuan Province. The soil samples were sampled from the top 0-20cm soil layer, and 100mg / kg Pb was added. 2+ . The Pb 2+ The addition of is realized by the metal solution, the metal solution is Pb(NO 3 ) 2 . Thoroughly mix the metal solution with the soil and passivate for three months. Then biochar is added to the soil polluted by heavy metals, and the amount of biochar is 5% of the soil mass. Weigh 2 kg of mixed soil samples and place them in plastic basins with an upper diameter of 20.5 cm and a height of 12 cm. The biochar used in this example is rice husk biochar.

[0059] First grow seedlings in seedling pots. When planting, choose pakchoi with no significant difference in appearance, plant 3 to 4 plants in each pot, and take 8mL of γ-PGA with different concent...

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Abstract

The invention belongs to the field of microorganisms, and in particular relates to a high-yielding γ-PGA bacillus subtilis and its application. The main technical scheme is as follows: a new strain of Bacillus subtilis is provided, the preservation number is: CGMCC NO.17215; its 16Sr DNA sequence is shown in SEQ ID NO 1. And based on the bacillus subtilis, an application method for improving soil environment and promoting plant growth is provided. Using the Bacillus subtilis provided by the present invention and its application method, high-yield γ-PGA can be obtained at a lower temperature, which greatly saves production energy consumption; effectively improves the growth performance of plants and reduces the absorption of heavy metals in soil by plants .

Description

technical field [0001] The invention belongs to the field of microbes, and in particular relates to a high-yield gamma-PGA bacillus subtilis and its application. Background technique [0002] γ-polyglutamic acid (English name poly-γ-glutamic acid, referred to as γ-PGA) is a naturally occurring water-soluble homopolyamide, with a large number of free carboxyl groups on its side chain, which can adsorb metal cations. The study found that the metal ions adsorbed by γ-PGA are mainly Ni 2+ 、Cd 2+ , Pb 2+ 、Cu 2+ , Mn 2+ 、Cr 2+ Wait. Based on this, γ-PGA can not only be used as a functional polymer for environmental governance, but also can concentrate radionuclides and recover heavy metals from solutions. [0003] Pb is highly toxic and non-degradable, can accumulate in the soil, and will cause harm to the human body through the food chain, so heavy metal pollution has attracted widespread attention from scholars. In recent decades, excessive fertilization, improper dispos...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12P13/02A01N63/22A01P21/00C12R1/125
CPCC12N1/20C12P13/02C12R2001/125C12N1/205Y02A40/22Y02P60/21
Inventor 闫志英李海红赵琪琪许力山姬高升刘杨
Owner CHENGDU INST OF BIOLOGY CHINESE ACAD OF S
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