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Phage microcapsule and preparation method thereof

A bacteriophage and microcapsule technology, applied in the field of microbial preparations, can solve problems such as difficult to achieve controlled release, achieve good acid resistance and enteric solubility, good enteric coating effect, and improve the effect of survival number

Active Publication Date: 2022-04-29
QINGDAO PHAGEPHARM BIO TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, there are still many immaturities in the above-mentioned embedded phage microcapsules, and some key issues have yet to be resolved, such as how to improve the encapsulation efficiency and storage efficiency of phage, and the difficulty in achieving controlled release.

Method used

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  • Phage microcapsule and preparation method thereof
  • Phage microcapsule and preparation method thereof
  • Phage microcapsule and preparation method thereof

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preparation example Construction

[0035] Preparation of bacteriophage fermentation broth: Take the stock solution of Escherichia coli phage, Salmonella phage, Vibrio alginolyticus phage, and Vibrio parahaemolyticus phage that have been separated and purified in a refrigerator at -80°C, and mix them with their host bacteria at a volume ratio of 1:1 Inoculated in the corresponding liquid nutrient broth, cultured on a shaker at 37°C and 170rpm for 5 hours, and centrifuged and filtered to detect the phage titer, the titer was 1.00×10 8pfu / mL~3.00×10 9 pfu / mL or so; then a large amount of phage was amplified according to the corresponding MOI to prepare a sufficient amount of phage, and the titers of different phages were respectively 2.00×10 9 pfu / mL~2.00×10 11 pfu / mL for future experiments.

[0036] 1. Exploration of the preparation method of phage microcapsules

[0037] 1. Preliminary exploration of the preparation method of phage microcapsules

[0038] (1) Experimental method:

[0039] The coli phage, Salm...

Embodiment 1

[0069] The present embodiment provides a kind of bacteriophage microcapsule, the preparation method of the bacteriophage fermented liquid that it adopts is the same as the aforementioned content, and its preparation method is as follows:

[0070] 1) Glue preparation: Weigh 5.0g trehalose, 1.5g calcium carbonate, 0.4mL 20% gelatin, add to 100mL bacteriophage fermentation broth, stir until completely dissolved and mix well, then add 1.9g sodium alginate, in a magnetic stirrer Fully stir for 15 minutes, mix well;

[0071] 2) Preparation of solidified solution: weigh 3.0g of calcium chloride, add 200mL of distilled water, and stir until completely dissolved;

[0072] 3) Preparation of washing solution: weigh 3.5g of sodium chloride, add 500mL of distilled water, and stir until completely dissolved;

[0073] 4) Coating material preparation: 2g chitosan powder, add 450mL 0.5% (w / v) acetic acid solution to dissolve, adjust pH=5.0 with 4M NaOH, dilute to 500mL and prepare immediately...

Embodiment 2

[0084] The present embodiment provides a kind of bacteriophage microcapsule, the preparation method of the bacteriophage fermented liquid that it adopts is the same as the aforementioned content, and its preparation method is as follows:

[0085] 1) Glue preparation: Weigh 6.0g trehalose, 2.0g calcium carbonate, 0.7mL 20% gelatin, add to 100mL bacteriophage fermentation broth, stir until completely dissolved and mix well, then add 2.4g sodium alginate, in a magnetic stirrer Fully stir for 15 minutes, mix well;

[0086] 2) Preparation of solidified solution: weigh 3.8g of calcium chloride, add 200mL of distilled water, and stir until completely dissolved;

[0087] 3) Preparation of washing solution: weigh 4.0g of sodium chloride, add 500mL of distilled water, and stir until completely dissolved;

[0088] 4) Coating material preparation: 2g chitosan powder, add 450mL 0.5% (w / v) acetic acid solution to dissolve, adjust pH=5.0 with 4M NaOH, dilute to 500mL and prepare immediately. ...

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Abstract

The invention discloses a bacteriophage microcapsule and a preparation method thereof, the bacteriophage microcapsule is mainly prepared from the following components: a bacteriophage core material, a protective agent and a wall material, the wall material comprises one or more of sodium alginate, gelatin, carrageenan, maltodextrin, chitosan and hydroxymethyl cellulose; the protective agent comprises one or more of amphoteric starch, trehalose, skimmed milk, calcium carbonate, cane sugar and maltodextrin. The preparation method comprises the following steps: respectively adding a wall material and a protective agent into bacteriophage fermentation liquor in proportion, and fully and uniformly mixing to obtain bacteriophage glue liquor; extruding the bacteriophage glue solution into the curing agent solution through a hole sharpening method, and standing and curing to obtain a wet capsule; washing the wet capsules with a washing solution; and soaking the washed wet capsule in a film coating material solution for film coating treatment to obtain the bacteriophage microcapsule. The bacteriophage microcapsule has good acid resistance and storage stability and high slow release rate, and the survival rate of the bacteriophage is improved.

Description

technical field [0001] The invention relates to the technical field of microbial preparations, in particular to a phage microcapsule and a preparation method thereof. Background technique [0002] Phage is a type of bacterial virus. The virulent phage can quickly lyse the bacteria and cause the death of the host bacteria, which has the effect of rapid sterilization. Phages grow and multiply in host cells, which can cause the lysis of pathogenic bacteria and reduce the density of pathogenic bacteria, thereby reducing or avoiding the chance of infection or disease of pathogenic bacteria, and achieving the purpose of treating and preventing diseases, that is, phage therapy. This therapy has been widely used in fields such as veterinary medicine, agriculture and food microbiology. The domestic breeding industry, especially the chicken industry, is often plagued by intestinal diarrhea in livestock and poultry, which is mainly caused by pathogenic microorganisms such as Escherich...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A23K10/18A23K20/163A23K20/147A23K10/20A23K20/24A23K40/30C12N7/00B01J13/04C12R1/92
CPCA23K10/18A23K20/163A23K20/147A23K10/20A23K20/24A23K40/30C12N7/00B01J13/04C12N2795/00021Y02A50/30
Inventor 潘强任慧英张召佐孙虎芝庄盈婷石静
Owner QINGDAO PHAGEPHARM BIO TECH CO LTD
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