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Method for preparing lactobacillus acidophilus powder by freeze-drying in vacuum

A technology of vacuum freeze-drying and Lactobacillus acidophilus is applied in the field of preparing Lactobacillus acidophilus powder by vacuum freeze-drying, which can solve problems such as cell inactivation

Active Publication Date: 2011-02-02
HANGZHOU GRAND BIOLOGIC PHARMA INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, if the choice or addition ratio of the protective agent is inappropriate, it will also lead to the inactivation of a large number of cells

Method used

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  • Method for preparing lactobacillus acidophilus powder by freeze-drying in vacuum
  • Method for preparing lactobacillus acidophilus powder by freeze-drying in vacuum

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Culture Lactobacillus acidophilus in MRS medium for 24h, then centrifuge at 6000r / min for 10min at a rotating speed, collect the thalline, then wash the thalline twice with normal saline, centrifuge and discard the supernatant; The cells were prepared to be 8±1×10 10 After cfu / mL bacterial suspension, let it stand for 30-40min, then add protective agent II and mix evenly, wherein, the volume ratio of the protective agent II and protective agent I is 1:1, then, let it stand for 20-30min before loading In a freeze-drying tube, the height of the sample is 0.3-0.5cm, and then placed in a refrigerator at -35 ~ -40°C for 24 hours, then transferred to a vacuum freeze dryer, at a temperature of -56°C and a vacuum of Freeze-dried for 24 hours under the condition of 4-6Pa; finally, reconstituted with normal saline, counted on MRS solid medium, and determined the activity of bacteria powder and the survival rate of bacteria.

[0018] The formula of above-mentioned protective agen...

Embodiment 2

[0022] Embodiment 2 is identical with embodiment 1 step, just, in aseptic MRS substratum, at first add the compound antifreeze factor of microporous membrane sterilization, then cultivate Lactobacillus acidophilus, wherein, the prescription of the protective agent I of adding It is: trehalose 10g / L+glycerin 20mL / L+bovine serum albumin 10g / L+sodium chloride 8.5g / L+sucrose 80g / L; the formula of the added protective agent II is: skim milk powder 150g / L+glucose 10g / L+sodium chloride 8.5g / L+ ascorbic acid 0.2g / L, the formula of described compound antifreeze factor is as shown in table 2 below:

[0023] Table 2 The verification results of the lyophilized survival rate of the bacteria obtained by adding different ratios of compound antifreeze factors fermented

[0024]

Embodiment 3

[0026] The method for preparing Lactobacillus acidophilus by vacuum freeze-drying in Example 3 is exactly the same as in Example 2, wherein, the formula of the compound antifreeze factor adopts: inositol 0.1g / L, sodium chloride 3g / L, bovine serum albumin 0.07g / L, alanine 0.03g / L; the formula of protective agent I adopts: trehalose 10g / L+glycerol 20mL / L+bovine serum albumin 10g / L+sodium chloride 8.5g / L+sucrose 80g / L; the formula of protective agent II Use: skimmed milk powder 150g / L+glucose 10g / L+sodium chloride 8.5g / L+ascorbic acid 0.2g / L, the volume ratio of the protective agent I and protective agent II is shown in the following table 3:

[0027] Table 3 The verification results of the survival rate of Lactobacillus acidophilus bacterial powder prepared by freeze-drying of the different volume ratios of the protective agent I and the protective agent II of the bacterial suspension

[0028] Protective agent I made into bacterial suspension: Protective agent II

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Abstract

The invention discloses a method for preparing lactobacillus acidophilus powder by freeze-drying in vacuum. The method comprises the following steps of: adding lactobacillus acidophilus into a culture medium for culturing the lactobacillus acidophilus; after centrifugal collection of thalli, washing the thalli with physiological saline and centrifuging to remove clear liquid; after preparing the thalli into bacterial suspension by using a protecting agent I; adding a protecting agent II; and finally performing freeze-drying, wherein the formula of the protecting agent I comprises the following components: 10 to 20g / L trehalose, 10 to 20 g / L glycerol, 5 to 15 g / L bovine serum albumin, 6.5 to 8.5g / L sodium chloride and 60 to 100 g / L sucrose; and the formula of the protecting agent II comprises the following components: 150 to 250g / L skim milk powder, 10 to 40g / L glucose, 6.5 to 8.5g / L sodium chloride, and 0.2 to 0.3g / L ascorbic acid. The survival rate of the lactobacillus acidophilus powder prepared by the invention is up to 99.9 percent.

Description

technical field [0001] The invention relates to a method for preparing Lactobacillus acidophilus powder by vacuum freeze-drying. Background technique [0002] With the development of the dairy market, especially the gradual enhancement of people's health awareness, the safety of food, health care products and medicine has attracted more and more attention. Microecological preparations containing probiotics are gradually being researched and developed as a new field. As a kind of probiotics, Lactobacillus acidophilus plays an important role in promoting human health and preventing diseases, and has increasingly become a hot spot in dairy scientific research. Lactobacillus acidophilus powder has been widely used in food, medicine and other industries, and has become an important modern industrial raw material [0003] Vacuum freeze-drying (abbreviated as freeze-drying) is one of the most effective methods for preparing and preserving biological materials, and freeze-drying i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/04C12R1/23
Inventor 陈合王磊马齐舒国伟秦涛
Owner HANGZHOU GRAND BIOLOGIC PHARMA INC
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