High-density culture method of bifidobacterium lactis and preparation method of freeze-dried powder of bifidobacterium lactis

A bifidobacterium, high-density fermentation technology, applied in the field of high-density fermentation and the preparation of freeze-dried bacterial powder, can solve the problems of large differences in the highest bacterial concentration, no definition of high-density culture, etc., to achieve the inhibition of pathogenicity Bacteria, reduced mortality, strong acid and bile salt tolerance

Active Publication Date: 2014-07-02
BEIJING SCITOP BIO TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the large difference in the maximum concentration of bacteria that can be achieved by various strains, there is no exact definition of high-density culture in the current literature.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] High-density fermentation of Bifidobacterium animalis subsp.lactis V9 (Bifidobacterium lactis V9):

[0044] a. Activation of strains: inoculate Bifidobacterium animalis subsp.lactis V9 (Bifidobacterium lactis V9, Bifidobacterium lactis V9) frozen at -40°C in 5ml TPY liquid medium (TPY medium: lactose 10g, beef extract 5g , yeast powder 5g, casein peptone 10g, soybean peptone 5g, KH 2 PO 4 2.5g, K 2 HPO 4 2.5g, MgSO 4 0.1g, Tween-800.25g, distilled water 1000mL, pH7.0, sterilized at 121°C for 15 minutes), anaerobic culture at 37°C for 20 hours, subcultured twice in this way to obtain activated strains.

[0045] b. Prepare optimized medium, lactose 8.0kg, beef extract 2.0kg, yeast powder 7.0kg, casein peptone 8.0kg, soybean peptone 7.0kg, KH 2 PO 4 1.0kg, K 2 HPO 4 4.0kg, MgSO 4 0.05kg, Tween-800.15kg, L-cysteine ​​hydrochloride 0.2kg, calcium carbonate 0.8kg, distilled water 1000L, fully dissolved.

[0046] c. Preparation of seed culture solution: inoculate 5% ...

Embodiment 2

[0049] High-density fermentation of Bifidobacterium animalis subsp.lactis V9 (Bifidobacterium lactis V9):

[0050] a. Activation of strains: inoculate Bifidobacterium animalis subsp.lactis V9 (Bifidobacterium lactis V9, Bifidobacterium lactis V9) frozen at -40°C in 5ml TPY liquid medium (TPY medium: lactose 10g, beef extract 5g , yeast powder 5g, casein peptone 10g, soybean peptone 5g, KH 2 PO 4 2.5g, K 2 HPO 4 2.5g, MgSO 4 0.1g, Tween-800.25g, distilled water 1000mL, pH7.0, sterilized at 121°C for 15 minutes), anaerobic culture at 37°C for 20 hours, subcultured twice in this way to obtain activated strains.

[0051] b. Prepare optimized medium, lactose 12.0kg, beef extract 5.0kg, yeast powder 5.0kg, casein peptone 12.0kg, soybean peptone 5.0kg, KH 2 PO 4 4.0kg,K 2 HPO 4 2.0kg, MgSO 4 0.15kg, Tween-800.35kg, L-cysteine ​​hydrochloride 0.5kg, calcium carbonate 1.2kg, distilled water 1000L, fully dissolved.

[0052] c. Preparation of seed culture solution: inoculate 8%...

Embodiment 3

[0055] Preparation of Bifidobacterium animalis subsp.lactis V9 (Bifidobacterium lactis V9) freeze-dried powder:

[0056] Centrifuge the high-density fermentation broth of Lactobacillus bifidobacterium V9 at 10,000g to obtain concentrated bacteria, and add the protective agent solution according to the ratio of concentrated bacteria to protective agent solution (10kg of skim milk, 8kg of lactose, 1kg of vitamin C, and 1kg of sodium glutamate , distilled water 1000L), mixed evenly, put into freeze-dryer and freeze-dry for 30 hours to obtain Bifidobacterium Lactobacillus V9 freeze-dried bacteria powder. The number of viable bacteria in the powder can reach 6.0×10 11 Above cfu / g, it can be directly applied to probiotic starters or probiotic preparations.

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Abstract

The invention relates to a high-density culture method of bifidobacterium lactis V9 and a preparation method of a freeze-dried powder of the bifidobacterium lactis V9, and belongs to the field of probiotics fermentation. The bifidobacterium lactis V9 (Bifidobacterium animalis subsp.lactis V9) is acid-resistant and cholate-resistant probiotics which are separated from the intestinal tract of a healthy Mongolian child, and the collection number of the bifidobacterium lactis V9 is CGMCC No.5470. High-density fermentation of the bacteria of the bifidobacterium lactis V9 is realized under the control of an optimized culture medium and optimized fermentation conditions; the viable count in a fermentation liquor can be more than 2.0*10<10>cfu/ml, and the viable count in the freeze-dried powder of the bifidobacterium lactis prepared by using the fermentation liquor can be more than 6.0*10<11>cfu/g, and therefore, the freeze-dried powder of the bifidobacterium lactis can be applied to a probiotic leavening agent or a probiotic preparation.

Description

technical field [0001] The invention relates to a high-density fermentation method of lactic acid bacteria and a preparation method of freeze-dried bacterial powder thereof, in particular to a high-density fermentation of bifidobacterium V9 and a preparation method of freeze-dried bacterial powder thereof. It belongs to the field of probiotic fermentation. technical background [0002] Lactobacillus bifidum is closely related to human life. It is a lactic acid bacteria commonly found in cream, meat and many vegetable fermented products. It can pass through the stomach and colonize the intestinal tract to play a beneficial role. It has an important impact on intestinal microbes, and has a wide range of applications in food fermentation, industrial lactic acid fermentation, and medical care. [0003] The bifidobacterium V9 used in the present invention is a strain of probiotics isolated and screened from the intestinal tract of healthy Mongolian children. Studies have shown ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12R1/01
Inventor 高鹏飞赵旭张善亭
Owner BEIJING SCITOP BIO TECH CO LTD
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