Method for high-density culture of ready-to-use lactobacillus plantarum, freeze-dried powder prepared by method and application

A technology of Lactobacillus plantarum and high-density culture is applied in the field of high-density culture method of Lactobacillus plantarum and the prepared freeze-dried powder, which can solve the problem that the safety and drug resistance of strain metabolites are unclear, and the strain has not been evaluated for safety. , unsuitable conditions and other problems, to achieve the effect of promoting a large number of growth, shortening the fermentation cycle, and reducing the number of expansion links

Inactive Publication Date: 2017-12-08
SOUTH CHINA UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The conditions of high-density medium, centrifugation, harvest age, protective agent, pre-freezing and other conditions selected by the above technology are not suitable, and the safety evalu...

Method used

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  • Method for high-density culture of ready-to-use lactobacillus plantarum, freeze-dried powder prepared by method and application
  • Method for high-density culture of ready-to-use lactobacillus plantarum, freeze-dried powder prepared by method and application
  • Method for high-density culture of ready-to-use lactobacillus plantarum, freeze-dried powder prepared by method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1: Safety Evaluation Research of Freeze-dried Powder

[0038] (1) The bacterial suspension of Lactobacillus plantarum DMDL9010 in the logarithmic phase growth is placed in the seed medium, and the activation culture is continuously carried out 2 to 3 times to obtain the seed culture liquid, and the bacterial content of the seed starter is 10 9 CFU / mL.

[0039] (2) Insert the seed starter into the high-density medium with a volume ratio of 3:100, carry out high-density culture at 37°C for 18 hours, and collect the fermentation suspension; the high-density medium is: calculated by mass , 1.0 parts of casein digest, 0.5 parts of yeast extract powder, 2.0 parts of glucose, 0.1 parts of triammonium citrate, 0.054 parts of magnesium sulfate, 0.8 parts of beef extract, 0.1 parts of dipotassium hydrogen phosphate, 0.5 parts of sodium acetate, Tween 80 0.18 parts, 1 part of soybean protein peptide, 0.02 parts of ascorbic acid, 0.15 parts of manganese sulfate, distilled ...

Embodiment 2

[0059] Example 2: Research on high-density cultivation of Lactobacillus plantarum DMDL9010

[0060] The first step is to put the Lactobacillus plantarum DMDL9010 bacteria powder in the MRS medium for 2 times of activation and collect the fermentation medium; put the fermentation medium into the high-density medium at a ratio of 3:100 by volume, and store it at 37°C Carry out fermentation culture, and collect the fermentation bacteria suspension after 18 hours; in parts by weight, the high-density medium is: 1.0 parts of casein digest, 0.5 parts of yeast extract powder, 2.0 parts of glucose, 0.2 parts of triammonium citrate , 0.058 parts of magnesium sulfate, 1.0 parts of beef extract, 0.1 parts of dipotassium hydrogen phosphate, 0.5 parts of sodium acetate, 0.18 parts of Tween 80, 1 part of soybean protein peptide, 0.02 parts of ascorbic acid, 0.1 parts of manganese sulfate, and distilled water to 100 parts, adjust the pH to 6.8. Stir and dissolve evenly, sterilize at 0.10MPa...

Embodiment 3

[0127] Preparation optimization research of embodiment 3 freeze-dried powder

[0128] The first step: the Lactobacillus plantarum DMDL9010 bacteria powder is placed in the MRS medium for 2 times of activation, and the fermentation medium is collected;

[0129] The second step: with a volume ratio of 3:100, insert the fermentation culture liquid into a high-density medium, carry out fermentation culture at 37°C, and collect the fermentation bacteria suspension after 18 hours; in parts by weight, the The high-density medium is: 1.0 parts of casein digest, 0.5 parts of yeast extract powder, 2.0 parts of glucose, 0.2 parts of triammonium citrate, 0.058 parts of magnesium sulfate, 1.0 parts of beef extract, 0.1 parts of dipotassium hydrogen phosphate, sodium acetate 0.5 part, Tween 80 0.18 part, soybean protein peptide 1 part, ascorbic acid 0.02 part, manganese sulfate 0.1 part, distilled water to 100 parts, adjust pH to 6.8. Stir to dissolve evenly, sterilize at 0.10MPa for 15min...

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Abstract

The invention discloses a method for high-density culture of ready-to-use lactobacillus plantarum, freeze-dried powder prepared by the method and application. The method comprises the following steps: (1) placing freeze-dried powder of lactobacillus plantarum in an MRS culture medium for activation twice to obtain a seed starter, wherein the microbial content of the seed starter is 108-109 CFU/mL; (2) inoculating a high-density culture medium with the seed starter according to the volume ratio of (3-6):100 for fermentation culture at 36-38 DEG C, and collecting fermentation bacterial suspension 18-24 h later; (3) centrifugating the fermentation bacterial suspension at 6000-8000 rpm for 25-35 min, discarding supernatant, washing sediment with sterile saline, and collecting bacterial sludge; (4) adding a composite protective agent to the bacterial sludge according to the volume ratio of the composite protective agent to the bacterial sludge of (2-5): 1, and obtaining bacterial suspension containing the protective agent after uniform mixing; and (5) pre-freezing the bacterial suspension containing the protective agent at -20 DEG C to -65 DEG C for 1.5-6.0 h, and then conducting drying in a vacuum freeze-dryer for 18-24 h to obtain the freeze-dried powder. The viable count of the prepared freeze-dried powder is as high as 1010-1011 cfu/g, the moisture content of the freeze-dried powder is less than 3%, and the freeze-dried powder is not prone to agglomeration, has good rehydration performance and can be widely used.

Description

technical field [0001] The invention relates to a direct injection type high-density cultivation method for plantar lactobacillus, in particular to a high-density cultivation method for plantaractobacillus and the prepared freeze-dried powder and its application. Background technique [0002] The high-density culture technology of lactic acid bacteria avoids the possibility that higher concentrations of initial substrates may inhibit the growth of microorganisms and reduce the final concentration. At present, domestic research on the high-density cultivation of Lactobacillus plantarum is developing, and its industrial application is still in the preliminary stage. Scholars have studied buffer salts and adding chemical neutralizers to promote the growth of lactic acid bacteria, but the concentration of the cultured bacterial suspension is still 10 9 cfu / mL levels. Other scholars have studied cell cycle culture and dialysis culture in order to obtain a large number of lactic...

Claims

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Application Information

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IPC IPC(8): C12N1/20A23L29/00A23C9/12A23G9/36C12R1/25
CPCA23C9/1234A23G9/363A23L29/065C12N1/20A23V2400/169
Inventor 刘冬梅黄燕燕何嘉敏赵珊林钟培周钦育
Owner SOUTH CHINA UNIV OF TECH
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