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Preparation method of bdellovibrio bacteriovorus lyophilized powder

A technology of phage leech vibrio and freeze-dried powder is applied in the field of biological preparations, which can solve the problems of weak leech vibrio infection ability, unknown freeze-drying effect, lack of reference, etc., to improve speed and stability, and is beneficial to The effect of protecting the environment and improving the survival rate of freeze-drying

Inactive Publication Date: 2017-09-29
XIAMEN CANCO BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Bdellovibrio cultured with an inactivated host has a weak infection ability, and at the same time, there is a potential environmental hazard for inactivation with chloroform, and at the same time, the host is Shigella flexneri or Escherichia coli, which have strong pathogenicity. If the inactivation is not complete, it will be very destructive to the environment
At the same time, the composition of the lyoprotectant is simple, and the lyophilization survival rate is not mentioned in the patent. The lyophilization effect is unknown, and there are obvious uncertainties and lack of reference

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1 The production process of Bdellovibrio lyophilized powder preparation 1

[0044] a. Obtaining the suspension of Bdellovibrio bacteria.

[0045] Routine culture method: get 50ml of Bdellovibrio liquid (10 7 pfu / ml) and 50ml Bacillus licheniformis liquid (10 8 cfu / ml) mixed, inoculated in a 20-liter fermenter with a liquid volume of 10 liters, a temperature of 28° C., a rotation speed of 150 rpm, and an air flow of 10 L / L / min. Fermented for 3 days to obtain Bdellovibrio fermentation broth

[0046] Bdellovibrio fermentation broth culture formula: 1% peptone, 0.3% beef extract, 0.5% sodium chloride, 0.3% yeast powder.

[0047] After the end of fermentation, add 1 liter of Bacillus subtilis bacterial liquid (10 13 cfu / ml), to ensure that the ratio of bacteriophage Bdellovibrio to Bacillus subtilis is 1:10.

[0048] b. Preparation of protective agent solution and preparation of lyophilized solution

[0049] The preparation process of the protective agent soluti...

Embodiment 2

[0053] Example 2 The production process of Bdellovibrio lyophilized powder preparation 2

[0054] a. Obtaining the suspension of Bdellovibrio bacteria.

[0055] Obtain bacterial liquid according to the following conventional culture method of Bdellovibrio bacteriophage

[0056] Routine culture method: get 100ml of Bdellovibrio liquid (10 7 pfu / ml) and 100ml Bacillus licheniformis liquid (10 8 cfu / ml) mixed, inoculated in a 30-liter fermenter, with a liquid volume of 20 liters, a temperature of 28° C., a rotation speed of 150 rpm, and an air flow of 20 L / L / min. Fermented for 3 days to obtain Bdellovibrio fermentation broth

[0057] Bdellovibrio fermentation broth culture formula: 1% peptone, 0.3% beef extract, 0.5% sodium chloride, 0.3% yeast powder.

[0058] After fermentation finishes, add 1 liter of Bacillus licheniformis bacterial liquid (10 13 cfu / ml), to ensure that the ratio of bacteriophage Bdellovibrio to Bacillus licheniformis is 1:100.

[0059] b. Preparation of...

Embodiment 3

[0065] Example 3 The production process of Bdellovibrio lyophilized powder preparation 3

[0066] a. Obtaining the suspension of Bdellovibrio bacteria.

[0067] According to the conventional culture method of Bdellovibrio bacteriophage, the bacterial liquid was obtained

[0068] Routine culture method: get 50ml of Bdellovibrio liquid (10 7 pfu / ml) and 50ml Enterococcus faecalis bacteria liquid (10 8 cfu / ml) were mixed, inoculated in a 10-liter fermenter, the liquid volume was 5 liters, the temperature was 28° C., the rotating speed was 100 rpm, and the ventilation rate was 5 L / L / min. Fermented for 3 days to obtain Bdellovibrio fermentation broth.

[0069] Bdellovibrio fermentation broth culture formula: 1% peptone, 0.3% beef extract, 0.5% sodium chloride, 0.3% yeast powder.

[0070] b. Preparation of protective agent solution and preparation of lyophilized solution

[0071] The preparation process of the protective agent solution is to dissolve skimmed milk powder, β-cyclo...

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Abstract

The invention provides a preparation method of a powdery bdellovibrio bacteriovorus lyophilized preparation with high survival rate. The method comprises the steps of obtaining a bacteria solution or a bdellovibrio bacteriovorus bacterial suspension from which a harmful host is separated according to a conventional culture method of bdellovibrio bacteriovorus; preparing a protective agent solution; carrying out freeze drying on a to-be-lyophilized mixed solution, repeatedly vacuumizing and filling an inert gas to finally prepare the preparation into powder; and carrying out sealing packaging on the bdellovibrio bacteriovorus lyophilized preparation under a vacuum condition or under the condition of filling the inert gas to prepare a finished product, namely the bdellovibrio bacteriovorus lyophilized preparation. Through compounding of multiple lyophilized protective agents, the freeze-drying survival rate of bdellovibrio bacteriovorus is improved, the character of the bdellovibrio bacteriovorus lyophilized preparation is powder, and dispersion and compounding of the lyophilized preparation are facilitated.

Description

technical field [0001] The invention belongs to the field of biological preparations, and in particular relates to a preparation method of Bdellovibrio lyophilized powder. Background technique [0002] The use of bioengineering technology to prevent and treat animal bacterial diseases has been widely used. Especially the application of Bdellovibrio has received more and more attention. Because bacteriophage Bdellovibrio mainly cracks Gram-negative bacteria, such as Escherichia coli, Salmonella, Aeromonas hydrophila, Vibrio and other Gram-negative pathogenic bacteria, and is harmless to humans and animals, compared with traditional The physical chemistry of antibiotics has clear advantages in eliminating pathogenic bacteria. [0003] Traditional physical and chemical antibiotics have obvious disadvantages in eliminating harmful bacteria. Physical methods can only be applied to certain links in related fields, and cannot cover the entire field. After long-term use of chemic...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/741A61K35/742A61K35/744A61K35/745A61K35/747A61K47/46A61K9/19A61P31/04
CPCA61K9/0019A61K9/19A61K35/741A61K35/742A61K35/744A61K35/745A61K35/747A61K47/46A61K2300/00
Inventor 郭立敬科举林茂
Owner XIAMEN CANCO BIOTECH CO LTD
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