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Pediococcus pentosaceus derived from breast milk and application of pediococcus pentosaceus

A technology of Pediococcus pentosacea and breast milk, which is applied in the field of Pediococcus pentosacea derived from breast milk and its application, can solve the problems of narrow antibacterial spectrum and poor antibacterial effect

Active Publication Date: 2020-10-20
SHIHEZI UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Among them, the patent CN109504637A discloses a strain of Pediococcus pentosaceae B49, which has a significant antibacterial effect on Escherichia coli aureus; Wang Yanting of Shanghai Ocean University isolated Pediococcus pentosaceae C-2-1 in Mongolian cheese, which has a significant antibacterial effect on Escherichia coli. Listeria has a strong inhibitory effect; patent 2018109565087 discloses a Pediococcus pentosaceae L1, which has a certain bacteriostatic effect on enterotoxigenic Escherichia coli, but the bacteriostatic effect of this bacterium is poor (bacteriostatic Circle > 6mm); patent WO2019174002A1 discloses a Pediococcus pentosaceae CCFM1012, which has a good antibacterial effect on Campylobacter jejuni
However, the Pediococcus pentosaceae disclosed in the above-mentioned patents only have antibacterial effects on individual individuals, and the antibacterial spectrum is narrow, and the antibacterial effect is relatively poor.

Method used

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  • Pediococcus pentosaceus derived from breast milk and application of pediococcus pentosaceus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1 Isolation and identification of Pediococcus pentosaceus HM75-1 derived from human milk

[0035] 1.1 Separation and purification

[0036] The breast milk samples (breast milk of Uyghur women in Kashgar, Xinjiang) were diluted with sterile water, and the dilution gradient was 10 -3 、10 -4 、10 -5 ; MRS medium (containing cysteine ​​0.5g / L) and Lactobacillus selective medium (LB medium) are sterilized and poured onto the plate, and after coagulation, 100 μL of breast milk samples of different dilution gradients are sucked and spread on the plate, Number and record, and invert for 24-48 hours in an anaerobic box at 37°C.

[0037] Select an appropriate gradient plate for each sample to pick 15-20 single colonies with different color, size and shape to observe the cell morphology, streak the colonies with rod-shaped or short rod-shaped cells for anaerobic culture, and repeatedly streak to The color, size and shape of the colonies in the same plate are consistent....

Embodiment 2

[0061] Example 2 Bacteriostatic properties of human milk source Pediococcus pentosaceus HM75-1

[0062] 2.1 Determination of antibacterial activity of strain HM75-1

[0063] Activation of strains: Take Pediococcus pentosaceus HM75-1 from breast milk that has been preserved in advance and ablate it, insert 1% into the sterilized MRS culture medium in an aseptic operating table, and incubate anaerobically at 37°C for 32 hours. Make the bacterial liquid in the logarithmic growth phase; take the food-borne pathogenic bacteria (diarrheogenic Escherichia coli (CICC 10411), Salmonella typhimurium (CICC 10420), enterotoxigenic Escherichia coli ( CICC 10421), hemorrhagic Escherichia coli (CICC21530), Listeria monocytogenes (CGMCC 1.9136), serotype Salmonella enteritidis (CGMCC1.10754)), ablation, press 1% The amount was added to the sterilized LB culture medium, and incubated at a constant temperature of 37°C for 24h. Make the bacterial liquid in the logarithmic growth phase.

[006...

Embodiment 3

[0073] Example 3 Optimization of culture conditions for human milk-derived Pediococcus pentosaceus HM75-1

[0074] 3.1 Effect of culture time on the antibacterial activity of strain HM75-1

[0075] Put the activated human milk-derived Pediococcus pentosaceus HM75-1 into the prepared MRS liquid medium according to the 10% inoculation amount, place it in an anaerobic incubator at 37°C for 52 hours, and take the bacterial solution every 4 hours. The growth curve is drawn by measuring the OD600nm value and pH value, the fermentation broth at each time point is centrifuged at 8000r / min for 15min, and the supernatant is collected for subsequent use; draw 150 μL of activated enterotoxigenic Escherichia coli (CICC 10421) bacteria suspension evenly spread on LB medium, let it stand for 30 minutes, and then put 4 sterilized Oxford cups (Oxford cup inner diameter is 8mm), and inject 200 μL of breast milk-derived pentose tablets at different time points into 3 of them One of the fermenta...

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Abstract

The invention belongs to the field of microbial fermentation, and particularly relates to pediococcus pentosaceus derived from breast milk and application of the pediococcus pentosaceus. The pediococcus pentosaceus derived from the breast milk is pediococcus pentosaceus HM75-1 and is preserved in the China Center for Type Culture Collection on October 22, 2019, and the preservation number of the pediococcus pentosaceus is CCTCC NO: M 2019849. The strain is high in bacteriostatic ability, has relatively high inhibitory activity on common pathogenic bacteria in intestinal tracts, and particularly has a remarkable inhibitory effect on enterotoxigenic escherichia coli; the strain is good in gastrointestinal tolerance and high in adaptability to gastrointestinal tracts of human bodies, and canbe finally used for preparing medicines for inhibiting pathogenic bacteria of intestinal tracts and preventing and treating food-borne diseases, infant milk powder supplements, food preservatives andthe like.

Description

technical field [0001] The invention belongs to the field of microbial fermentation, and in particular relates to a breast milk-derived Pediococcus pentosacea and an application thereof. [0002] technical background [0003] With the gradual improvement of national living standards in recent years, people pay more and more attention to food safety issues. Foodborne diseases are the most prominent problem at present, and in the field of food processing, food preservation and preservation also have a certain impact on food safety. The current additives have a long shelf life, but the cost of using physical methods is high and the effect is not good. And the scope of application is small. Therefore, the research on natural additives is becoming more and more extensive. [0004] Among them, Pediococcus pentosaceus belongs to the Streptococcus family Pediococcus, facultative anaerobic, Gram-positive bacteria, because it can ferment glucose to produce lactic acid, it is a kind o...

Claims

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Application Information

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IPC IPC(8): C12N1/20A61K35/741A61P31/04A23L33/135A23L3/3571A23C9/16A23C9/152C12R1/01
CPCC12N1/20A61K35/741A61P31/04A23L33/135A23L3/3571A23C9/16A23C9/152A23V2002/00C12R2001/01C12N1/205A23V2400/427A23V2200/10A23V2200/32A23V2200/30Y02A50/30
Inventor 张艳倪永清王艺璇罗宝龙徐梦
Owner SHIHEZI UNIVERSITY
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