Method for continuously culturing bacillus subtilis and special fermenting system

The technology of a Bacillus subtilis and its cultivation method is applied to the cultivation of Bacillus subtilis and the field of special equipment in the cultivation process, which can solve the problems of low spore formation rate and high production cost, achieve stable and uniform quality, prolong life, damage reduction effect

Active Publication Date: 2017-05-10
INST OF AGRI RESOURCES & ENVIRONMENT SHANDONG ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, in the industrial production process, there are mainly problems such as low sporulation rate and high production cost.

Method used

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  • Method for continuously culturing bacillus subtilis and special fermenting system

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] A fermentation system dedicated to the continuous culture method of Bacillus subtilis, 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 sequentially connected in series 8. Spore culture tank 9;

[0066] A seed tank 7 is connected in parallel between the cooling device 11 and the primary culture tank 6;

[0067] The spore culture tank 9 is connected with an auxiliary material tank 10;

[0068] A delivery pump I4 is connected between the batching tank 1 and the continuous elimination device 2; a delivery pump II12 is connected between the maintenance tank 3 and the cooling device 11;

[0069] 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.

[0070] Concrete steps are, inoculate 0.5-1% Bacillus subtilis bacterial cla...

Embodiment 2

[0074] The continuous culture method of bacillus subtilis comprises the following steps:

[0075] (1) Seed cultivation

[0076] a. Activation of strains on test tube slant

[0077] Transfer the preserved Bacillus subtilis strains to the test tube slant medium, keep at 33°C, and cultivate for 30 hours to activate the strains;

[0078] The formula of the 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;

[0079] b. Erlenmeyer flask seed culture

[0080] Pick 1-2 rings of Bacillus subtilis strains from the activated test tube and inoculate them into the Erlenmeyer flask, keep at 33°C, 120rpm after inoculation, and cultivate for 17 hours;

[0081] The medium formula of the triangular flask is: glucose 20g / L, peptone 15g / L, NaCl 5g / L, beef extract 0.5g / L, pH7.1, sterilized at 121°C for 30 minutes;

[0082] c. Seed tank culture

[0083] Inoculate the culture material A in the seed tank b...

Embodiment 3

[0105] The continuous culture method of bacillus subtilis comprises the following steps:

[0106] (1) Seed cultivation

[0107] a. Activation of strains on test tube slant

[0108] Transfer the preserved Bacillus subtilis strains to the test tube slant medium, keep at 32°C, and cultivate for 24 hours to activate the strains;

[0109] 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;

[0110] b. Erlenmeyer flask seed culture

[0111] Pick 1-2 rings of bacteria from the activated test tube and inoculate them into the Erlenmeyer flask, keep at 32°C, 100rpm after inoculation, and cultivate for 16 hours;

[0112] The medium formula of the Erlenmeyer flask is: glucose 20g / L, peptone 15g / L, NaCl 5g / L, beef extract 0.5g / L, pH 7.0, sterilized at 121°C for 30 minutes;

[0113] c. Seed tank culture

[0114] Inoculate the culture material A in the seed tank by inoculating the ab...

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Abstract

The invention belongs to the field of microbes, and particularly relates to a method for continuously culturing bacillus subtilis and a special fermenting system. The method provided by the invention comprises the following steps: (1) transferring stored bacillus subtilis strain to a test tube agarslantculture-medium, performing strain activation, and then, transferring the strain into a triangular flask and a seeding tank for culturing in sequence to prepare a bacillus subtilis seed; (2) taking culture materials including starch, corn powder, soya bean meal, peptone, NaCl, CaCl2, MgSO4.7H2O, KH2PO4, K2HPO4, MnSO4, a defoaming agent and water; (3) after the culture materials in the step (2) are subjected to sterilizing treatment, inoculating the activated and cultured bacillus subtilis seed, and performing three-stage continuous culturing to obtain a fermenting liquid. The bacillus subtilis cultured with the method provided by the invention is stable and uniform in quality; neither mixed fungi pollution nor strain mutation is easily caused; the service life of various detectors is prolonged; the time is reduced; the yield and the production efficiency are improved; the concentration and the spore rate of thalli of the bacillus subtilis are improved.

Description

technical field [0001] The invention belongs to the field of microorganism culture, and in particular relates to a method for cultivating bacillus subtilis, and also relates to special equipment for the above-mentioned culturing process of bacillus subtilis. Background technique [0002] Bacillus subtilis is an aerobic bacillus that widely exists in nature. It has strong lipase, protease, amylase, cellulase and other enzymatic activities. It has strong metabolism, is harmless to humans and animals, and does not pollute the environment. . In recent years, Bacillus subtilis, as a safe, efficient, multifunctional and highly potential microbial strain, has been widely used in the fields of industry, agriculture, medicine and health, food, animal husbandry, aquatic products and scientific research. [0003] Bacillus is a dormant body with extremely low water content and strong stress resistance formed in the cell when the vegetative body of bacteria is about to run out of essent...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12M1/34C12M1/36C12M1/00C12R1/125
CPCC12M23/58C12M41/12C12M41/48C12N1/20
Inventor 任海霞宫志远万鲁长韩建东李瑾任鹏飞黄春燕谢红艳曲玲
Owner INST OF AGRI RESOURCES & ENVIRONMENT SHANDONG ACADEMY OF AGRI SCI
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