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A method for high-density continuous fermentation of Clostridium butyricum and a preparation method of Clostridium butyricum microecological preparation

A technology of Clostridium butyricum and a fermentation method, applied in the field of microbial fermentation, can solve problems such as cumbersome steps, and achieve the effects of avoiding cumbersome operations, reducing negative feedback, and reducing the accumulation of metabolites

Active Publication Date: 2020-09-01
XIAMEN CANCO BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Patent CN103710286B is a secondary seed fermentation for Clostridium butyricum, the steps are cumbersome, and the fermentation period is 28-32 hours at the same time, and the fermentation density is up to 2×10 9 cfu / ml

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] (1) Seed cultivation:

[0059] Prepare 300ml of RCM seed medium, each 1L of seed medium contains: yeast extract 2g, beef extract 8g, peptone 8g, soluble starch 0.5g, glucose 5g, cysteine ​​hydrochloride 0.5g, NaCl 3g, NaAc 3g , resazurin 3mg, the pH value of the seed medium is 6.0. The prepared seed medium was sterilized under high pressure at 121°C for 20 minutes.

[0060]The Clostridium butyricum seed spore liquid was transferred to the seed medium, placed in an anaerobic chamber protected by nitrogen for cultivation, and cultivated at 37°C for 18 hours to obtain the Clostridium butyricum seed liquid.

[0061] (2) Continuous fermentation

[0062] Prepare fermentation medium and feed medium, and carry out 121 ° C, high-pressure sterilization for 20 minutes, wherein the specific medium components are as follows:

[0063] Fermentation medium: peptone: 10g / L, sugarcane honey: 40g / L; MgSO 4 0.2g / L; K 2 HPO 4 4g / L, KH 2 PO 4 1g / L, MnSO 4 1mg / L;

[0064] Feed medi...

Embodiment 2

[0070] (1) Seed cultivation:

[0071] Prepare 400ml of RCM seed medium, each 1L of seed medium contains: yeast extract 3g, beef extract 10g, peptone 12g, soluble starch 1g, glucose 5g, cysteine ​​hydrochloride 0.5g, NaCl 3g, NaAc 3g, Resazurin 3 mg, the pH value of the seed medium is 6.0, and the prepared seed medium is subjected to high-pressure sterilization at 121° C. for 20 minutes.

[0072] The Clostridium butyricum seed spore liquid was transferred to the seed medium, placed in an anaerobic chamber protected by nitrogen for cultivation, and cultivated at 37°C for 15 hours to obtain the Clostridium butyricum seed liquid.

[0073] (2) Continuous fermentation

[0074] Prepare fermentation medium and feed medium, and carry out 121 ° C, high-pressure sterilization for 20 minutes, wherein the specific medium components are as follows:

[0075] Fermentation medium: peptone: 25g / L, lignocellulosic hydrolyzed syrup: 50g / L; MgSO 4 0.2g / L; K 2 HPO 4 4g / L, KH 2 PO 4 1g / L, MnS...

Embodiment 3

[0082] (1) Seed cultivation:

[0083] Prepare 500ml of RCM seed medium, each 1L medium seed medium contains: yeast extract 4g, beef extract 12g, peptone 12g, soluble starch 1.5g, glucose 6g, cysteine ​​hydrochloride 0.5g, NaCl 3g, NaAc 3g , resazurin 3mg, the pH value of the seed medium is 6.0, and the prepared seed medium is sterilized under high pressure at 121° C. for 20 minutes.

[0084] The Clostridium butyricum seed spore liquid was transferred to the seed medium, placed in an anaerobic chamber protected by nitrogen for cultivation, and cultivated at 37°C for 12 hours to obtain the Clostridium butyricum seed liquid.

[0085] (2) Continuous fermentation

[0086] Prepare fermentation medium and feed medium, and carry out 121 ° C, high-pressure sterilization for 20 minutes, wherein the specific medium components are as follows:

[0087] Fermentation medium: peptone: 20g / L, beet honey: 40g / L; MgSO 4 0.2g / L; K 2 HPO 4 4g / L, KH 2 PO 4 1g / L, MnSO 4 1mg / L;

[0088] Fee...

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Abstract

The invention relates to the field of microbiological fermentation, and discloses a method for continuously fermenting clostridium butyricum under high density and a preparation method for a clostridium butyricum micro-ecological preparation. The preparation method comprises the following steps: adopting multi-stage continuous fermentation for a first time, dropping fermentation liquor in a lastformer-stage fermentation tank into a next-stage fermentation tank at a speed of 5-10% of the volume of the fermentation tank per hour, supplementing a fermentation culture medium or a supplementary culture medium, controlling concentration of residual sugar in the fermentation tank of each stage until residual sugar in the last-stage fermentation tank is lower than 15% or below, and discharging fermentation liquor to obtain a clostridium butyricum bacterial solution; and vacuum mixing and drying the clostridium butyricum bacterial solution and a carrier in vacuum to obtain the clostridium butyricum micro-ecological preparation. The method for continuously fermenting clostridium butyricum under high density can control bacteria growth rate and bacteria density in the fermentation tank at each stage, reduces accumulation of metabolite to the greatest extent, and promotes further growth at the later fermentation period of clostridium butyricum; and concentration of the obtained clostridiumbutyricum bacterial solution is higher than 10<11>cfu / ml.

Description

technical field [0001] The invention relates to the field of microbial fermentation, in particular to a method for high-density continuous fermentation of Clostridium butyricum, and also includes a method for continuously fermenting and producing high-density Clostridium butyricum probiotics. Background technique [0002] Clostridium butyricum, Clostridium butyricum, also known as Clostridium butyricum, is an anaerobic Gram-negative bacterium widely present in the intestinal tract, yogurt and soil of animals or humans. Clostridium butyricum can form endophytic spores, and has the characteristics of high temperature resistance, gastric acid resistance, bile salt resistance, and resistance to some antibiotics. At the same time, Clostridium butyricum can stimulate the immune system and improve the body's immunity. Its metabolites have certain anti-cancer properties, promote the self-repair of small intestinal cells and improve the absorption function of small intestinal cells. ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N1/04C12R1/145
CPCC12N1/04C12N1/20
Inventor 郭立敬科举林茂
Owner XIAMEN CANCO BIOTECH CO LTD
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