A kind of continuous culture method of Bacillus licheniformis and special fermentation system

A technology of Bacillus licheniformis and culture methods, applied in the direction of microorganism-based methods, specific-purpose bioreactors/fermenters, biochemical equipment and methods, etc., capable of solving problems such as death and poor stress resistance of vegetative bodies

Active Publication Date: 2020-12-01
INST OF AGRI RESOURCES & ENVIRONMENT SHANDONG ACADEMY OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, due to the relatively poor vegetative stress resistance of Bacillus licheniformis, it is easy to die in large numbers in the process of making products, and the spores formed in the late growth period are different from vegetative cells in structure and chemical composition, so they are also Has many properties different from vegetative cells

Method used

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  • A kind of continuous culture method of Bacillus licheniformis and special fermentation system

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Experimental program
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Effect test

Embodiment 1

[0064] A special fermentation system for the continuous culture method of Bacillus licheniformis, including a batching tank 1, a continuous elimination device 2, a maintenance tank 3, a cooling device 11, a feed liquid storage tank 5, a primary culture tank 6, and a secondary culture tank connected in series in sequence 8. Spore culture tank 9;

[0065] A seed tank 7 is connected in parallel between the cooling device 11 and the primary culture tank 6;

[0066] The spore culture tank 9 is connected with an auxiliary material tank 10;

[0067] The delivery pump delivery pump I4 is connected between the batching tank 1 and the continuous elimination device 2; the delivery pump delivery pump II12 is connected between the maintenance tank 3 and the cooling device 11;

[0068] There is a constant flow pump between the material storage tank, the primary cultivation tank 6, the secondary cultivation tank 8 and the spore cultivation tank 9.

[0069] The specific steps are to inocula...

Embodiment 2

[0071] The continuous culture method of Bacillus licheniformis comprises the following steps:

[0072] (1) Seed cultivation

[0073] a. Activation of strains on test tube slant

[0074] Transfer the preserved Bacillus licheniformis strains to the test tube slant medium, keep at 33°C, and cultivate for 30 hours to activate the strains;

[0075] The formula of test tube slant medium is: glucose 4g / L, peptone 3g / L, NaCl 1g / L, beef extract 0.5g / L, agar 20g / L, pH7.1, sterilized at 121°C for 20 minutes;

[0076] b. Erlenmeyer flask seed culture

[0077] Pick 1-2 rings of Bacillus licheniformis strains from the activated test tube and inoculate them into the Erlenmeyer flask, keep at 33°C, 120rpm after inoculation, and cultivate for 19 hours;

[0078] The medium formula of the triangular flask is: glucose 15g / L, soybean meal powder 16g / L, peptone 1g / L, NaCl 1g / L, KH 2 PO 4 0.3g / L, K 2 HPO 4 0.3g / L, pH7.1, sterilized at 121°C for 20 minutes;

[0079] c. Cultivation of 7 strai...

Embodiment 3

[0099] The continuous culture method of Bacillus licheniformis comprises the following steps:

[0100] (1) Seed cultivation

[0101] a. Activation of strains on test tube slant

[0102] Transfer the preserved Bacillus licheniformis strains to the test tube slant medium, keep at 32°C, and cultivate for 24 hours to activate the strains;

[0103] The formula of test tube slant medium is: glucose 4g / L, peptone 3g / L, NaCl 1g / L, beef extract 0.5g / L, agar 20g / L, pH7.0, sterilized at 121°C for 20 minutes;

[0104] b. Erlenmeyer flask seed culture

[0105] Pick 1-2 rings of activated Bacillus licheniformis strains from the test tube and inoculate them into the Erlenmeyer flask, and culture them for 20 hours at 32°C and 100 rpm after inoculation;

[0106] The medium formula of the triangular flask is: glucose 15g / L, soybean meal powder 16g / L, peptone 1g / L, NaCl 1g / L, KH 2 PO 4 0.3g / L, K 2 HPO 4 0.3g / L, pH7.0, sterilized at 121°C for 20 minutes:

[0107] c. Cultivation of 7 stra...

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Abstract

The invention belongs to the field of microbes, and particularly relates to a Bacillus licheniformis continuous culture method and a special fermentation system. The method comprises the following steps: (1) inoculating a Bacillus licheniformis strain into a test tube slant culture medium, activating the strain, sequentially inoculating into a triangular flask a seeding tank, and culturing, thereby preparing the Bacillus licheniformis seeds; (2) taking a culture medium composed of starch / glucose, corn flour, bean pulp powder, (NH4)2HPO4, NaCl, CaCl2, MgSO4.7H2O, KH2PO4, K2HPO4, MnSO4, a defoaming agent and water; and (3) sterilizing the culture medium in the step (2), inoculating and activating the cultured Bacillus licheniformis seeds, and carrying out three-stage continuous culture, thereby obtaining the final fermentation liquid. The method implements high mechanization and automation, and the Bacillus licheniformis has stable and uniform quality and can not have the phenomena of infectious microbe pollution and strain variation. The method prolongs the service lives of detecting instruments, saves the time, enhances the yield and production efficiency, and increases the Bacillus licheniformis strain concentration and spore rate.

Description

technical field [0001] The invention belongs to the field of microorganism cultivation, and in particular relates to a continuous cultivation method of Bacillus licheniformis, and also relates to special equipment in the above-mentioned cultivation process of Bacillus licheniformis. Background technique [0002] Bacillus licheniformis is one of the more potential strains of Bacillus. Belonging to Gram-positive rod-shaped bacteria, it can produce endogenous spores. It is heat-resistant and stress-resistant. It is ubiquitous in soil and plant surfaces. It is a common endophyte in plants and is non-toxic and harmless to humans and animals. [0003] Bacillus licheniformis can produce a variety of antibacterial substances. Under certain conditions, Bacillus licheniformis can produce lipopeptides, peptides, phospholipids, polyenes and amino acids and other antibiotics, which have a good effect on a variety of plant pathogens. Inhibitory effect, Bacillus licheniformis can produce ...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12M1/34C12M1/00C12R1/10
CPCC12M23/58C12M41/00C12M41/12C12M41/48C12N1/20
Inventor 任海霞宫志远万鲁长任鹏飞韩建东张海兰杨鹏郭兵姚强
Owner INST OF AGRI RESOURCES & ENVIRONMENT SHANDONG ACADEMY OF AGRI SCI
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