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A kind of breast milk source Pediococcus pentosacea and its application

A technology for Pediococcus pentosaceus and breast milk, which is applied to Pediococcus pentosaceus from breast milk and its application field, can solve the problems of poor bacteriostatic effect and narrow bacteriostatic spectrum, and achieves good gastrointestinal tolerance and strong bacteriostatic activity. , the effect of improving immunity

Active Publication Date: 2022-05-20
SHIHEZI UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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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  • A kind of breast milk source Pediococcus pentosacea and its application
  • A kind of breast milk source Pediococcus pentosacea and its application
  • A kind of breast milk source Pediococcus pentosacea and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1 Isolation and identification of Pediococcus pentosaceus HM75-1 derived from human milk

[0033] 1.1 Separation and purification

[0034] 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.

[0035] 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

[0059] Example 2 Bacteriostatic properties of human milk source Pediococcus pentosaceus HM75-1

[0060] 2.1 Determination of antibacterial activity of strain HM75-1

[0061] 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 pathogens preserved in advance (enteropathogenic Escherichia coli (CICC 10411), Salmonella enterica serotype Typhimurium (CICC 10420), Enterotoxigenic Escherichia coli (CICC 10421), Enterohaemorrhagic Escherichia coli (CICC 21530), Listeria monocytogenes (CGMCC 1.9136), Salmonella enterica subsp. enterica (CGMCC 1.10754) , ablated, put 1% into sterilized LB culture medium in a sterile operating table, and incubated at 37°C for 24 hours. Make the bacterial liquid in the logarit...

Embodiment 3

[0071] Example 3 Optimization of culture conditions for human milk-derived Pediococcus pentosaceus HM75-1

[0072] 3.1 Effect of culture time on the antibacterial activity of strain HM75-1

[0073] 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 in particular relates to a breast milk-derived Pediococcus pentosacea and an application thereof. The breast milk-derived Pediococcus pentosaceus is Pediococcus pentosaceus HM75‑1, which was deposited in the China Center for Type Culture Collection on October 22, 2019, with a preservation number of CCTCC NO: M 2019849. The bacterial strain has strong antibacterial ability, and has strong inhibitory activity on common intestinal pathogenic bacteria, especially has significant inhibitory effect on enterotoxigenic Escherichia coli; and the bacterial strain has better gastrointestinal tolerance, It has strong adaptability to the human gastrointestinal tract, and can eventually be used to prepare drugs for inhibiting intestinal pathogenic bacteria, prevent and treat food-borne diseases, infant milk powder supplements, food preservatives, etc.

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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Patent Type & Authority Patents(China)
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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