A kind of Enterococcus faecium, its high-density fermentation culture method and microecological preparation prepared therefrom

A high-density fermentation, Enterococcus faecium technology, applied in microorganisms, microorganisms, preservation microorganisms and other directions, can solve the heat resistance of Enterococcus faecium, gastric acid resistance, bile salt resistance and fermentation performance can not adapt, bacterial drug resistance drug residues aggravate and other problems, and achieve the effect of significant probiotics and stress resistance, biological characteristics of bile salt tolerance, and not easy to absorb water.

Active Publication Date: 2020-02-14
QINGDAO VLAND BIOTECH INC +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, the heat resistance, gastric acid resistance, bile salt resistance and fermentation performance of the existing Enterococcus faecium cannot meet the needs of the growing industrialization, and cannot meet the needs of the modern feed industry and animal husbandry. Therefore, screening for probiotics and Enterococcus faecium strains with significant stress resistance have become a major bottleneck in the development of animal husbandry
In addition, the continuous feeding of antibiotics has led to the increasing resistance of bacteria and drug residues in livestock products.

Method used

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  • A kind of Enterococcus faecium, its high-density fermentation culture method and microecological preparation prepared therefrom
  • A kind of Enterococcus faecium, its high-density fermentation culture method and microecological preparation prepared therefrom

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Embodiment 1

[0027] The present embodiment provides a new Enterococcus faecium, whose isolation, screening and identification are as follows:

[0028] Under aseptic conditions as far as possible, 1 g of the intestinal content of the rectum of healthy animals was collected with a sterile spoon, and placed in 30 mL of MRS medium (formulated peptone 10.0 g / L, beef powder 8.0 g / L, yeast powder 4.0 g / L). L, Glucose 20.0g / L, K 2 HPO 4 2.0g / L, diamine hydrogen citrate 2.0g / L, sodium acetate 5.0g / L, MgSO 4 0.2g / L, MnSO 4 0.04g / L, Tween-80 1.0g / L. ) in a 100mL Erlenmeyer flask, enriched and cultured for 8h, then drew 1mL and 9mL sterile saline to prepare a 1:10 dilution (i.e. prepared into 10 -1 diluent), vibrate for 3-5min, use a micropipette to accurately draw 1mL of the diluent into a 9 mL sterilized saline test tube, vibrate with a vortex shaker for 1-2min, and prepare 10 -2 The dilution solution, followed by 10 -3 -10 -6 Dilution, select 10 -3 -10 -6 For three dilutions, draw 200 μ...

Embodiment 2

[0037] The present embodiment provides a kind of high-density fermentation culture method of the Enterococcus faecium isolated and identified as above-mentioned embodiment, specifically as follows:

[0038] Fermentation culture method for primary production seeds:

[0039] Take the basal seeds of Enterococcus faecium E13-5 and inoculate the slant of MRS agar medium under sterile conditions for pure culture. The test tubes are cultured statically at 37°C-40°C for 24-48 hours. After passing the inspection of colony observation and microscopic examination, etc., they are used as first-class production seeds and stored at 2-8°C, and the storage time is generally not more than 1 month;

[0040] Secondary production seed fermentation culture method:

[0041] Take the primary production seed slant of Enterococcus faecium E13-5, add 4mL sterile normal saline to the slant of the strain to wash the colonies on the slant, and inoculate it into the secondary seed culture solution accordi...

Embodiment 3

[0051] This embodiment provides a preparation process of a microecological preparation prepared from Enterococcus faecium E13-5 isolated and identified in the above embodiment as the main raw material, specifically as follows:

[0052] Get a predetermined amount of Enterococcus faecium E13-5 and centrifuge under the condition of rotating speed 10000r / min to obtain the bacteria sludge, the slag discharge period is 50-60min, then mix the bacteria sludge with the protective agent (in parts by mass, the Protective agent comprises 5% sucrose, 4% sorbitol, 3% skimmed milk powder, 1% gelatin and 0.5% Vc,) carry out thorough mixing with the ratio of mass ratio 1:3, then carry out lyophilization successively under the following conditions: at -40 Pre-freeze at ℃ for 3-4 hours, vacuumize to about 20Pa at -40℃ to -45℃, sublimate and dry at 5℃ for 20-25 hours, sublimate and dry at 0℃ for 8-10 hours, analyze and dry at 25℃ for 4- 6 hours, finally close the vacuum for 2-4 hours, and freeze-...

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Abstract

The invention proposes an Enterococcus faecium, its high-density fermentation method and a microecological preparation prepared therefrom, which belong to the technical field of lactic acid bacteria cultivation. Enterococcus faecium strains with significant probiotic and stress resistance are screened out as feed additives, which can effectively Replace some antibiotics. The Enterococcus faecium provided by the technical solution is E13-5, and the preservation number is CCTCC M2017191. The enterococcus faecium provided by the invention can be used as a safe and efficient new feed additive to partially replace antibiotics, thereby meeting the requirements of the modern feed industry and animal husbandry.

Description

technical field [0001] The invention belongs to the technical field of lactic acid bacteria cultivation, and in particular relates to a kind of Enterococcus faecium, its high-density fermentation and cultivation method and the probiotics prepared therefrom. Background technique [0002] Enterococcus faecium belongs to the Streptococcus family, Enterococcus bacteria, Gram-positive, facultative anaerobic. It has excellent biological characteristics and is an intestinal symbiotic bacterium that can form a dominant flora in the intestinal tract. Due to its fast growth rate, good adhesion, and the production of lactic acid and some antibacterial substances, it is found in the newborn 2 of many animals. -Have a dominant position within 3 days, thereby increasing the number of beneficial bacteria, inhibiting harmful bacteria, promoting intestinal health, regulating the micro-ecological balance of the intestinal tract and preventing diarrhea. It has a significant effect on improving...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N1/04A23K10/18C12R1/01
CPCA23K10/18C12N1/04C12N1/20C12N1/205C12R2001/01
Inventor 马丰英蒋贻海王艳玲周英俊崔栩王宏华武利利孙亚磊刘元元魏波
Owner QINGDAO VLAND BIOTECH INC
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