Protection agent applied to feeding enterococcus faecium freeze-drying preparation and preparation method of protection agent

A technology of Enterococcus faecium and protective agent, applied in the field of microorganisms, can solve the problems of strict storage conditions of freeze-dried preparations, less research and application, and few Lactococcus to develop excellent freeze-dried preparations, etc.

Inactive Publication Date: 2015-04-29
BEIJING DAWEIJIA BIOTECH SHARE CO LTD +1
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  • Abstract
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AI Technical Summary

Problems solved by technology

[0004] Most of the viable counts of currently commercially available freeze-dried lactic acid bacteria preparations are 2×10 11 -5×10 11 CFU/g range, rarely more than 5×10 11 The level of CFU/g, and most of the products belong to human products, the research and application of the method of vacuum freeze-drying in the production of live la

Method used

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  • Protection agent applied to feeding enterococcus faecium freeze-drying preparation and preparation method of protection agent
  • Protection agent applied to feeding enterococcus faecium freeze-drying preparation and preparation method of protection agent
  • Protection agent applied to feeding enterococcus faecium freeze-drying preparation and preparation method of protection agent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Embodiment 1, the single factor optimization experiment of protective agent composition

[0026] 1. Selection and preparation of protective agent components:

[0027] Prepare 7% skimmed milk powder solution, 1% maltodextrin, galacto-oligosaccharide, sucrose, lactose, trehalose, maltose and glycerin solution, 0.1% sodium glutamate, glycine and lysine solution , 2% sorbitol and mannitol solution, after sterilization, carry out single factor test (wherein the amino acid protective agent is sterilized by filtration, and the sterilization condition of other protective agents is 115°C, 20min).

[0028] 2. Collection of bacteria slime:

[0029] High-density fermented Enterococcus faecium (Enterococcus faecium) WEI-10 CGMCC No.7746 fermentation broth was centrifuged at 10,000-15,000 rpm to collect the sludge through a tubular centrifuge, and the centrifugation rate was controlled at 40-60 L / h.

[0030] 3. Take 30mL of the sterilized protective agent solution above and 6g of E...

Embodiment 2

[0038] Example 2. Response Surface Optimization Experiment of Protective Agent Components

[0039] Response surface test design can integrate the interaction between multiple factors in the test results, and use statistical software to fit the quadratic function model equation to find the best optimization plan. 46 groups of experiments with 5 factors and 3 levels were designed by Box-Behnken design method, with concentration of 5 significant influencing factors as independent variable and survival rate as response value. According to the single factor test results, the coding levels of the 5 factors are shown in Table 2. The Box-Behnken test design and results of 46 groups of 5 factors and 3 levels are shown in Table 3, among which 40 groups of tests are factorial points, and the remaining 6 groups of tests are zero points. The independent variable takes the center point, which is used to estimate the experimental error. The collection of bacteria slime, mixing with differen...

Embodiment 3

[0057] Embodiment 3, the influence of different mixing ratios of protective agent and bacteria slime on freeze-drying effect

[0058] 1. According to the optimal protective agent formula, mix the bacteria sludge collected after fermentation and the protective agent according to the ratio (W / V) of 1:1, 1:3, 1:5, 1:10, and 1:20, respectively. Homogenize for 5 minutes at 2800rpm in a homogenizer.

[0059] 2. Take 36g of bacterial suspension and place them on stainless steel trays, pre-freeze at -80°C for 5 hours, then put them into a freeze dryer at -40°C for vacuum freeze-drying, and the freeze-drying time is 44 hours.

[0060] 3. After the freeze-dried solid was pulverized, the viable bacteria were counted, and the survival rate was calculated according to the method in Example 1.

[0061] 4. Analysis of results: by image 3 The results showed that when the ratio of the sludge to the protective agent was 1:3 and 1:5, the survival rate of about 90% could be obtained, which was...

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Abstract

The invention relates to a preparation method of a protection agent and a freeze-drying preparation for vacuum freeze-drying of feeding enterococcus faecium. By virtue of the preparation method of the freeze-drying protection agent and the freeze-drying preparation, a high-density fermentation solution of enterococcus faecium WEI-10CGMCC No.7746 is processed by vacuum freeze-drying, the maximum number of live bacteria of freeze-dried bacteria powder reaches 8.2*10<11> CFU/g; and the freeze-drying protection rate is 98%. The freeze-drying preparation can be preserved for one year at -20 DEG C, and the loss rate of the live bacteria is smaller than 5%; and the freeze-drying preparation can be preserved for 6 months at the room temperature, and the loss rate of the live bacteria is smaller than 10%. The preparation method can be used for producing high-yield and storable freeze-drying preparation of the feeding enterococcus faecium.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, and relates to a protective agent for vacuum freeze-drying treatment of enterococcus faecium for feeding and a preparation method of a freeze-dried preparation. Background technique [0002] Vacuum freeze-drying is a commonly used and effective method for long-term preservation of active lactic acid bacteria. However, during the freeze-drying process, the lactic acid bacteria cells will be damaged in various ways, mainly including the formation of ice crystals, the mechanical damage of the cell membrane, the change of the osmotic pressure of the cell membrane, the loss of water in the cell, the increase of the solute concentration, and the solute effect. , Sensitive protein denaturation and inactivation, the loss of water affects the function of many hydrophilic macromolecular substances in the cell, in addition, it will also lead to the destruction of the dynamic balance of pH inside and ...

Claims

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Application Information

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IPC IPC(8): C12N1/04C12R1/46
CPCC12N1/04
Inventor 乔琳姚宏明冯媛媛高长斌刘蕊金忠辉
Owner BEIJING DAWEIJIA BIOTECH SHARE CO LTD
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