CN110507598A - Lactic acid bacteria is inhibiting the application in propionibacterium acnes and skin care item made of it - Google Patents
Lactic acid bacteria is inhibiting the application in propionibacterium acnes and skin care item made of it Download PDFInfo
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- CN110507598A CN110507598A CN201910853475.8A CN201910853475A CN110507598A CN 110507598 A CN110507598 A CN 110507598A CN 201910853475 A CN201910853475 A CN 201910853475A CN 110507598 A CN110507598 A CN 110507598A
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- fermentation liquid
- parts
- supernatant
- tmc3115
- lactic acid
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Classifications
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- A—HUMAN NECESSITIES
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
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- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
- A61K35/745—Bifidobacteria
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- A—HUMAN NECESSITIES
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- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
- A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
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- A—HUMAN NECESSITIES
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Abstract
Lactic acid bacteria is inhibiting the application in propionibacterium acnes and skin care item made of it; belong to the technical field of lactic acid bacteria; the present invention provides can be effectively applied to the lactic acid bacteria thallus and/or lactic acid bacteria derivative for inhibiting propionibacterium acnes; and the Deacne pack made of lactic acid bacteria thallus and/or lactic acid bacteria derivative, acne eliminating cream and facial treatment essence liquid are provided, the lactic acid bacteria and/or lactic acid bacteria derivative can protect original skin barrier while inhibiting propionibacterium acnes.Fermentation liquid, supernatant, thallus and the lysate of LP45, TMC3115 and LR863 are significant to propionibacterium acnes fungistatic effect；Fermentation liquid, thallus and the lysate of LR519 is significant to propionibacterium acnes fungistatic effect；The supernatant and thallus of BAL531 is significant to propionibacterium acnes fungistatic effect, and skin-protection product is used for after being further processed；The dead bacterium of LR863 has fungistatic effect, and fungistatic effect is reduced with LR863 concentration and is deteriorated, and can directly choose suitable concentration as needed for skin care item such as facial masks.
Description
Technical field
The invention belongs to the technical fields of lactic acid bacteria, are related to the application that lactic acid bacteria inhibits bacterium, and in particular to lactic acid bacteria exists
Inhibit application in propionibacterium acnes and with its manufactured skin care item.
Background technique
Acne is a kind of chronic inflammatory skin of pilosebaceous follicle, and disease incidence 70%-87% is apt to occur in puberty,
Clinical manifestation is mainly with facial area and chest and back blackhead, Whiteheads, based on inflammatory papules, warts.Under androgenic effect
Sebaceous glands Rapid development and lipid largely secrete be acne occur pathophysiological basis；Pilosebaceous duct dyskeratosis is
Another key factor and main pathological phenomenon occur for acne, and epithelial cell angling makes pilosebaceous duct blocking, sebum discharge
Obstacle, ultimately forms visible micro mist thorn and clinical macroscopic acne under microscope, and micro mist thorn and acne are formed as
Propionibacterium acnes proliferation with anaerobic growth characteristic creates good local environment.Acne occur exactly with acne propionic acid
Bacillus is closely related, it is now recognized that propionibacterium acnes may be taken part in by the innate immunity, acquired immunity and directly induction
The occurrence and development of acne inflammation.Acne Earlier period of inflammation may be induction caused by the innate immune reaction that Toll-like is mediated by (TLR)
Proinflammatory inflammation factor especially IL-1 α release；As disease develops, the acquired immune response is exaggerated inflammatory process, further leads
Cause inflammatory factor release and neutrophil accumulation；Propionibacterium acnes also can produce polypeptides matter, directly induces or aggravates
Inflammation.In end-stage disease, follicular wall is broken, and the substances such as lipid, hair in hair follicle enter corium, and it is anti-that inflammation has been further aggravated
It answers.
Part patients with acne skin barrier is impaired, and Long-term Oral or external application anti-acne drug such as Tretinoin, often aggravates
The destruction of skin barrier, leads to skin sensitivity.Therefore, how effectively to inhibit propionibacterium acnes, while protecting original skin
Barrier is the emphasis that we research and develop at present.
Summary of the invention
The present invention can be effectively applied to the lactic acid bacteria thallus for inhibiting propionibacterium acnes to solve the above problems, providing
And/or lactic acid bacteria derivative, and provide the skin care item made of lactic acid bacteria thallus and/or lactic acid bacteria derivative, the lactic acid bacteria
And/or lactic acid bacteria derivative can protect original skin barrier while inhibiting propionibacterium acnes.
The present invention be realize its purpose the technical solution adopted is that: lactic acid bacteria inhibit propionibacterium acnes in application,
It is critical that the lactic acid bacteria is lactic acid bacteria thallus and/or lactic acid bacteria derivative；
Lactic acid bacteria thallus includes lactobacillus plantarum (Lactobacillus plantarum) LP45, CGMCC No.8072,
Lactobacillus acidophilus (Lactobacillus acidophilus) La28, CGMCC No.11506, bifidobacterium lactis
(Bifidobacteriumlactis) BAL531, CGMCC No.17329, bifidobacterium bifidum
(Bifidobacteriumbifidum) TMC3115, CGMCC No.8462, Lactobacillus rhamnosus (Lactobacillus
Rhamnosus) LR519, CGMCC No.15969, Lactobacillus rhamnosus (Lactobacillus rhamnosus) LR863,
One or more of CGMCC No.14410；
The lactic acid bacteria derivative include the fermentation liquid of above-mentioned LP45, LP45 inactivation fermentation liquid, LP45 supernatant,
LP45 inactivation supernatant, the lysate of LP45, the fermentation liquid of La28, La28 inactivation fermentation liquid, La28 supernatant,
La28 inactivation supernatant, the fermentation liquid of BAL531, the supernatant of BAL531, BAL531 inactivation supernatant, BAL531 it is molten
Born of the same parents' product, the fermentation liquid of TMC3115, TMC3115 inactivation fermentation liquid, the supernatant of TMC3115, TMC3115 inactivation supernatant
Liquid, the lysate of TMC3115, the fermentation liquid of LR519, LR519 inactivation fermentation liquid, the supernatant of LR519, LR519 it is molten
Born of the same parents' product, the fermentation liquid of LR863, LR863 inactivation fermentation liquid, the supernatant of LR863, LR863 inactivation supernatant, LR863
Lysate or one or more of the dead bacterium of LR863.
If above-mentioned lactic acid bacteria is inhibiting the application in propionibacterium acnes, the fermentation liquid of the LP45, La28 fermentation liquid,
The fermentation liquid of BAL531, the fermentation liquid of TMC3115, the fermentation liquid of LR519 or LR863 fermentation liquid prepared by following steps:
LP45, La28, BAL531, TMC3115, LR519 or LR863 by after fluid nutrient medium culture to logarithmic phase with 20%-30%'s
Glycerol etc. freezes to obtain seed liquor in -60 DEG C~-80 DEG C than being uniformly mixed；
Seed liquor is taken to be inoculated into 10ml fluid nutrient medium by the inoculum concentration of 2%-10%, concussion mixes, 30 DEG C of -40 DEG C of cultures
- 48h for 24 hours successively continuously activates three generations, then is inoculated into expand in 250ml fluid nutrient medium by the inoculum concentration of 2%-10% and cultivate,
Up to the fermentation liquid of LP45, the fermentation liquid of La28, the fermentation liquid of BAL531, the fermentation liquid of TMC3115, LR519 fermentation liquid or
The fermentation liquid of LR863；
The fermentation liquid of the fermentation liquid or TMC3115 of above-mentioned BAL531 is prepared as anaerobic condition；
The fermentation liquid of LP45, the fermentation liquid of La28, the fermentation liquid of TMC3115, LR519 fermentation liquid or LR863 fermentation
Liquid heats 10-20min at 110-130 DEG C to get the inactivation of the inactivation fermentation liquid of LP45, the inactivation fermentation liquid, TMC3115 of La28
Fermentation liquid, the inactivation fermentation liquid of LR519 or the inactivation fermentation liquid of LR863.
If above-mentioned lactic acid bacteria is inhibiting the application in propionibacterium acnes, the fermentation liquid of the LP45, La28 fermentation liquid,
The fermentation liquid 7000-9000r/min centrifugation of the fermentation liquid of BAL531, the fermentation liquid of TMC3115, the fermentation liquid of LR519 or LR863
After 10-15min, supernatant is collected to get the supernatant of LP45, the supernatant of La28, the supernatant of BAL531, TMC3115
The supernatant of supernatant, the supernatant of LR519 or LR863；
The supernatant of LP45, the supernatant of La28, the supernatant of BAL531, the supernatant of TMC3115 or the supernatant of LR863
Liquid heats 10-20min at 110-130 DEG C to get the inactivation of the inactivation supernatant of LP45, the inactivation supernatant, TMC3115 of La28
The inactivation supernatant of supernatant or LR863；
The fermentation liquid of the LP45, the fermentation liquid of La28, the fermentation liquid of BAL531, the fermentation liquid of TMC3115, LR519
After the fermentation liquid of fermentation liquid or LR863 centrifugation gained bacterium mud are sufficiently resuspended with 0.9% sodium chloride solution, 7000-9000r/ is repeated
Min be centrifuged 10-15min three times to get the thallus of LP45, the thallus of La28, the thallus of BAL531, TMC3115 thallus,
The thallus of LR519 or the thallus of LR863；
The fermentation liquid of the LR863 is with 60-80 DEG C of sterilizing 2-4h after 7000-9000r/min is centrifuged emulsification 10-15min
It is freeze-dried the dead bacterium to get LR863.
If above-mentioned lactic acid bacteria is inhibiting the application in propionibacterium acnes, the thallus of the LP45, BAL531 thallus,
Pasteur is gone out after the thallus of TMC3115, the thallus of the thallus of LR519 or LR863 and protective agent are mixed and stirred for uniformly according to 1:2
Bacterium to get the lysate of LP45, the lysate of BAL531, the lysate of TMC3115, LR519 lysate or
The lysate of LR863.
If above-mentioned lactic acid bacteria is inhibiting the application in propionibacterium acnes, further, according to the mass fraction, the protection
Agent includes 10-15 portions of skimmed milks, 2-6 portions of trehaloses, 1.5-2 parts of vitamin Cs, 1-3 parts of sodium glutamates, 60-80 parts of distilled water.
If above-mentioned lactic acid bacteria is inhibiting the application in propionibacterium acnes, further, according to the mass fraction, the liquid
Culture medium includes 10-20 parts of glucose, 5-10 parts of peptones, 2-5 parts of sodium acetates and 65-83 parts of distilled water.
If above-mentioned lactic acid bacteria is inhibiting the application in propionibacterium acnes, further, according to the mass fraction, the liquid
Culture medium includes 10-20 parts of glucose, 5-10 parts of peptones, 2-6.5 parts of powdered beefs, 1-5 parts of yeast extracts, 2-5 parts of sodium acetates,
0.5-2 parts of diammonium hydrogen citrates, 0.8-2 parts of dipotassium hydrogen phosphates, 0.32-0.58 parts of MgSO4·7H2O, 0.25-1 parts of MnSO4
H2O, 0.1-0.5 parts of cysteine hydrochlorides, 0.01-0.1 parts of Tween 80s, 800-1000 parts of distilled water.
It is a kind of using above-mentioned lactic acid bacteria inhibit in propionibacterium acnes using manufactured Deacne pack, Deacne pack by
Facial mask essence and face-mask material composition, according to the mass fraction, facial mask essence include 0.3 part of -0.6 part of lactic acid bacteria thallus and/
Or lactic acid bacteria derivative, 10-13 parts of glycerol, 0.04-0.08 parts of Sodium Hyaluronates and 86.32-89.45 parts of distilled water, facial mask material
Material includes one or more of non-woven fabrics, silk, biological fiber or tencel fiber.
It is a kind of using above-mentioned probiotics inhibit propionibacterium acnes in using manufactured acne eliminating cream, by mass fraction
Meter, acne eliminating cream include 0.4-0.7 parts of probio thallines and/or probiotic derived object, 4.8-5.5 parts of vaseline, and 3.3-3.84 parts
Octadecyl alcolol, 1.44-2.5 parts of monoglycerides, 2.72-2.98 parts of Valelinum Liquidums, 0.31-0.54 parts of lauryl sodium sulfate, 2.4-3.53
Part glycerol, 81.68-83 parts of distilled water.
It is a kind of using above-mentioned probiotics inhibit propionibacterium acnes in using manufactured facial treatment essence liquid, by mass parts
Number meter, facial treatment essence liquid includes 9-12 parts of probio thallines and/or probiotic derived object, 10-13.4 parts of glycerol, 0.05-0.09
Part Sodium Hyaluronate, 0.04-0.06 parts of niacinamide, 1-2 parts of triglycerides, 0.05-0.08 parts of EDETATE SODIUMs.
Lactobacillus plantarum (Lactobacillus plantarum) LP45, Merchant Codes lactobacillus plantarum LP45, preservation are compiled
Number be YMC1005, deposit number is CGMCC No.8072, and depositary institution is China General Microbiological culture presevation administrative center, protect
Hiding address is Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, and Institute of Microorganism, Academia Sinica, preservation date is 2013 8
The moon 26.
Lactobacillus acidophilus (Lactobacillus acidophilus) La28, deposit number is CGMCC No.11506, preservation
Unit is China General Microbiological culture presevation administrative center, and preservation address is Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3,
Institute of Microorganism, Academia Sinica, preservation date are on October 15th, 2015.
Bifidobacterium lactis (Bifidobacteriumlactis) BAL531, deposit number are CGMCCNo.17329, preservation list
Position is China General Microbiological culture presevation administrative center, and preservation address is Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, in
Institute of microbiology, the academy of sciences, state, preservation date are on March 13rd, 2019.
Bifidobacterium bifidum (Bifidobacteriumbifidum) TMC3115, deposit number are CGMCC No.8462, are protected
Hiding unit is China General Microbiological culture presevation administrative center, and preservation address is Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3
Number, Institute of Microorganism, Academia Sinica, preservation date is on November 11st, 2013.
Lactobacillus rhamnosus (Lactobacillus rhamnosus) LR519, deposit number is CGMCC No.15969, is protected
Hiding unit is China General Microbiological culture presevation administrative center, and preservation address is Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3
Number, Institute of Microorganism, Academia Sinica, preservation date is on June 20th, 2018.
Lactobacillus rhamnosus (Lactobacillusrhamnosus) LR863, deposit number are CGMCC No.14410, preservation
Unit is China General Microbiological culture presevation administrative center, and preservation address is Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3,
Institute of Microorganism, Academia Sinica, preservation date are on July 12nd, 2017.
Inhibiting the beneficial effects of the present invention are: lactic acid bacteria and/or lactic acid bacteria derivative can be used as a kind of mild bacteriostatic agent
Original skin barrier is protected while propionibacterium acnes.
Fermentation liquid, supernatant, thallus and the lysate of LP45, TMC3115 and LR863 are to the antibacterial effect of propionibacterium acnes
Fruit is significant；
And fermentation liquid, thallus and the lysate of LR519 are significant to propionibacterium acnes fungistatic effect；
And the supernatant and thallus of BAL531 are significant to propionibacterium acnes fungistatic effect, are used for after being further processed
Skin-protection product；
The dead bacterium of same LR863 has fungistatic effect, and fungistatic effect is reduced with LR863 concentration and is deteriorated, can basis
Need directly to choose suitable concentration for skin care item such as facial masks.
Detailed description of the invention
Fig. 1 is 1 streptococcus acidi lactici fermented solution fungistatic effect figure of embodiment.
Fig. 2 is 1 lactic acid bacteria of embodiment inactivation fermentation liquid fungistatic effect figure.
Fig. 3 is 1 lactic acid bacteria supernatant fungistatic effect figure of embodiment.
Fig. 4 is 1 lactic acid bacteria of embodiment inactivation supernatant fungistatic effect figure.
Fig. 5 is 1 lactic acid bacteria thallus fungistatic effect figure of embodiment.
Fig. 6 is 1 lactic acid bacteria of embodiment inactivation thallus fungistatic effect figure.
Fig. 7 is 1 lactic acid bacteria lysate fungistatic effect figure of embodiment.
Fig. 8 is the dead bacterium fungistatic effect figure of 1 lactic acid bacteria of embodiment.
A1-F1 is followed successively by the fermentation liquid fungistatic effect figure of LP45, La28, BAL531, TMC3115, LR519 and LR863；
A11-F11 is followed successively by the inactivation fermentation liquid fungistatic effect figure of LP45, La28, BAL531, TMC3115, LR519 and LR863；
A2-F2 is followed successively by the supernatant fungistatic effect figure of LP45, La28, BAL531, TMC3115, LR519 and LR863；A22—F22
It is followed successively by the inactivation supernatant fungistatic effect figure of LP45, La28, BAL531, TMC3115, LR519 and LR863；A3-F3 is successively
For the thallus fungistatic effect figure of LP45, La28, BAL531, TMC3115, LR519 and LR863；A33-F33 be followed successively by LP45,
The inactivation thallus fungistatic effect figure of La28, BAL531, TMC3115, LR519 and LR863；A4, C4-F4 be followed successively by LP45,
The lysate fungistatic effect figure of BAL531, TMC3115, LR519 and LR863；E, F is respectively the dead bacterium suppression of LR519, LR863
Bacterium effect picture, wherein 1x109CFU/ml、5x109CFU/ml indicates the concentration of dead bacterium；" 1 " indicates stoste in figure, and " 10 " indicate 10
Times dilution, " 100 " indicate 100 times of dilutions, and "+" indicates positive control, and "-" indicates negative control.
Specific embodiment
The present invention is further detailed with attached drawing combined with specific embodiments below.
One, it screens
Embodiment 1
1.1 stock
Propionibacterium acnes: purchase receives biology from north, number BNCC336649.
Protective agent: as mass fraction, including skimmed milk 15g, trehalose 4g, vitamin C 1g, sodium glutamate 1.5g are pure
Water 78.5g.
MRS liquid culture medium: including glucose 20g, peptone 10g, powdered beef 6.5g, yeast extract 5g, acetic acid
Sodium 5g, diammonium hydrogen citrate 2g, dipotassium hydrogen phosphate 2g, MgSO4·7H2O0.58g, MnSO4H2O 0.25g, cysteine salt
Hydrochlorate 0.5g, Tween 80 1ml, distilled water 1000mL.
Sulphur glycollate culture medium: including junket peptone 15.0g, yeast extract powder 5.0g, glucose 5.0g, sodium thioglycollate
0.5g, l-cysteine 0.5g, resazurin 0.001g, sodium chloride 2.5g, agar 0.75g, distilled water 1000mL, the pH value of culture medium
For 6.8-7.3.
Lactobacillus solution: isolate and purify to obtain from Yoghourt or baby intestinal original strain (lactobacillus plantarum LP45, it is thermophilic
Lactobacillus lactis La28, bifidobacterium lactis BAL531, bifidobacterium bifidum TMC3115, Lactobacillus rhamnosus LR519, rhamnose cream
Bacillus LR863), it is uniformly mixed after the improved MRS fluid nutrient medium culture to logarithmic phase of original strain with 30% glycerol grade ratio,
It is frozen in -80 DEG C up to seed liquor, lactobacillus solution is provided by Hebei Yiran Biological Technology Co., Ltd.'s strain library.
Further include 0.9% sodium chloride solution, the quadracycline of 30 μ g/ml, 34 μ g/ml quadracycline.
(1) fermentation liquid: taking lactobacillus solution to be inoculated into 10ml MRS liquid culture medium by 3% inoculum concentration, concussion
It mixes, 37 DEG C of cultures for 24 hours, successively continuously activate three generations, then be inoculated into 250ml MRS liquid culture medium by 3% inoculum concentration
Middle expansion is cultivated, wherein bifidobacterium lactis BAL531, bifidobacterium bifidum TMC3115 palpus Anaerobic culturel, other incubation step phases
Together；
Cultured fermentation liquid takes 100ml to get fermentation liquid bulk samples, dilutes stoste with 0.9% sodium chloride solution, obtains
10 times of dilution samples, it is spare in 4 DEG C of refrigerations；
(2) inactivate fermentation liquid: 121 DEG C of heating 15min of fermentation liquid must inactivate fermentation liquid bulk samples, with 0.9% sodium chloride
Solution dilutes stoste, obtains 10 times of dilution samples；
(3) supernatant: fermentation liquid 8000r/min is centrifuged 10min and collects supernatant to get supernatant bulk samples, uses
0.9% sodium chloride solution dilutes stoste, obtains 10 times of dilution samples, spare in 4 DEG C of refrigerations；
(4) inactivate supernatant: 121 DEG C of heating 15min of supernatant must inactivate supernatant bulk samples, with 0.9% sodium chloride
Solution dilutes stoste, obtains 10 times of dilution samples；
(5) thallus: centrifugation gained bacterium mud is centrifuged after being sufficiently resuspended with 0.9% sodium chloride solution, in triplicate up to thallus
Bulk samples dilute stoste with 0.9% sodium chloride solution, obtain 10 times of dilution samples, and 100 times of dilution samples are refrigerated in 4 DEG C
It is spare；
(6) inactivate thallus: 121 DEG C of heating 15min of thallus bulk samples must inactivate thallus bulk samples, with 0.9% chlorination
Sodium solution dilutes stoste, obtains 10 times of dilution samples, 100 times of dilution samples；
(7) lysate: pasteurization is after thallus and protective agent are mixed and stirred for uniformly according to 1:2 to get lysate
Sample dilutes stoste with 0.9% sodium chloride solution, obtains 10 times of dilution samples, 100 times of dilution samples；
(8) dead bacterium: fermentation liquid is freeze-dried with 70 DEG C of sterilizing 3h after 8000r/min is centrifuged emulsification 10min up to dead bacterium
Sample is configured to 5x10 with 0.9% sodium chloride solution and dead bacterium respectively8CFU/ml、1x109CFU/ml、5x109CFU/ml is dense
The fluid sample of degree.
(9) culture of propionibacterium acnes: propionibacterium acnes seed liquor is taken to be inoculated into 10ml liquid by 3% inoculum concentration
Sulphur glycollate culture medium, concussion mix, 37 DEG C of Anaerobic culturel 48h, successively continuously spare after activation three generations.
1.2 processing
(1) it is cooled to 50 DEG C or so containing 2.0% element agar culture medium, 10mL is taken to be poured in sterilized petri dishes, cleaning work
It is dried in platform；
(2) aseptic nipper takes sterilizing Oxford cup to be placed in the plate of element agar, stands 10min；
(3) 1mL propionibacterium acnes 100mL is added to be cooled in 50 DEG C of thioglycollate medium and fill
Divide and mixes；
(4) topple over 10mL aforesaid liquid sulphur glycollate culture medium on the plate for put well Oxford cup, it is to be solidified after by ox
Saliva cup presss from both sides out, forms Oxford cup aperture；
(5) 0.1mL each sample is injected in each Oxford cup aperture, in 37 DEG C of standing Anaerobic culturel 48h.And with 0.9%
Sodium chloride solution is negative control；Fermentation liquid, fire extinguishing fermentation liquid, supernatant and inactivation supernatant are with 30 μ g/ml quadracyclines
For positive control, thallus, inactivation thallus, lysate and dead bacterium are using 34 μ g/ml quadracyclines as positive control, such as Fig. 1-
8 be the fungistatic effect figure of lactic acid bacteria.
1.3 post-processing
Table 1
1.4 analysis
With the diameter of vernier caliper measurement inhibition zone, measurement result such as table 1, wherein evaluation index are as follows: inhibition zone is straight
Diameter=0 does not have fungistatic effect then；Antibacterial circle diameter > 0, then have fungistatic effect；0 < antibacterial circle diameter < 18, antibacterial effect
Fruit is general；18≤antibacterial circle diameter < 27mm, fungistatic effect are good；Antibacterial circle diameter >=27mm, fungistatic effect are significant；" -- " table
Show and does not do experiment.The selection result is as follows:
(1) the Oxford cup bacteriostatic test of fermentation liquid shows: the antibacterial circle diameter of LP45, TMC3115, LR519 and LR863 are big
In 30mm, fungistatic effect is significant, and the fungistatic effect of La28, BAL531 are good；By inactivation after LP45, La28, TMC3115,
LR519 and LR863 only has stoste to have fungistatic effect, and effect is general, and the inactivation fermentation liquid of BAL531 does not have fungistatic effect.
(2) the Oxford cup bacteriostatic test of supernatant shows: the supernatant of LP45, BAL531, TMC3115 and LR863 are antibacterial
Loop diameter about 30mm, fungistatic effect is significant, and the fungistatic effect of La28 is good, and only stoste has suppression after above-mentioned five kinds of bacterial strains inactivation
Bacterium effect, effect are general；LR519 only has stoste to have fungistatic effect, and effect is general.
(3) the Oxford cup bacteriostatic test of thallus shows: the antibacterial effect of LP45, BAL531, TMC3115, LR519 and LR863
Fruit is significant, and La28 only has stoste to have good fungistatic effect；Six kinds of bacterial strains do not have fungistatic effect after inactivation.
(3) the Oxford cup bacteriostatic test of lysate shows: the fungistatic effect of LP45, TMC3115, LR519 and LR863 are aobvious
It writes, wherein LR863 lysate fungistatic effect is best, and BAL531 only has stoste to have certain fungistatic effect, and La28 lysis produces
Object is without fungistatic effect.
(4) the Oxford cup bacteriostatic test of dead bacterium shows: in LR519 and LR863, only LR863 has fungistatic effect, and antibacterial effect
Fruit increases with LR863 concentration and is increased, and fungistatic effect is good when wherein concentration is 5,000,000,000, and 1,000,000,000 antibacterial circle diameters are 17.24mm,
Fungistatic effect is general.
To sum up, the fermentation liquid of LP45, TMC3115 and LR863, supernatant, thallus and lysate are to propionibacterium acnes
Fungistatic effect is significant；And fermentation liquid, thallus and the lysate of LR519 are also very aobvious to propionibacterium acnes fungistatic effect
It writes；There are also the supernatant of BAL531 and thallus are significant to propionibacterium acnes fungistatic effect, skin care is used for after being further processed
Product；
The dead bacterium of LR863 has fungistatic effect, and fungistatic effect is reduced with LR863 concentration and is deteriorated, can be straight as needed
Selecting takes suitable concentration for skin care item such as facial masks.
1.5 reference examples
Reference examples 1
Identical to treatment process as above-mentioned stock, difference, which is only that, replaces with Liquid Culture for MRS liquid culture medium
Base 1, according to the mass fraction, fluid nutrient medium 1 include 10 parts of glucose, 5 parts of peptones, 2 parts of sodium acetates and 83 parts of distilled water.
Reference examples 2
Identical to treatment process as above-mentioned stock, difference, which is only that, replaces with Liquid Culture for MRS liquid culture medium
Base 2, according to the mass fraction, fluid nutrient medium 2 include 15 parts of glucose, 8 parts of peptones, 4 parts of sodium acetates and 73 parts of distilled water.
Reference examples 3
Identical to last handling process as above-mentioned stock, difference, which is only that, replaces with liquid training for MRS liquid culture medium
Base 3 is supported, according to the mass fraction, fluid nutrient medium 3 includes 20 parts of glucose, 10 parts of peptones, 5 parts of sodium acetates and 65 parts of distillations
Water.
For culture medium described in above-mentioned reference examples 1-3 compared with MRS liquid culture medium, MRS liquid culture medium makes bacterium
Strain is growing more rapidly, and metabolite is more, and antibacterial circle diameter is bigger, more preferable to the inhibitory effect of propionibacterium acnes.
Embodiment 2
2.1 stock
Propionibacterium acnes, sulphur glycollate culture medium, 0.9% sodium chloride solution, 30 μ g/ml quadracycline,
The quadracycline of 34 μ g/ml is same as Example 1, and difference is only that MRS liquid culture medium and lactobacillus solution；
Protective agent: including 10 portions of skimmed milks, 2 portions of trehaloses, 1.5 parts of vitamin Cs, 1 part of sodium glutamate, 80 parts of distilled water.
MRS liquid culture medium: including glucose 10g, peptone 5g, powdered beef 2g, yeast extract 1g, sodium acetate
2g, diammonium hydrogen citrate 0.5g, dipotassium hydrogen phosphate 0.8g, MgSO4·7H2O 0.32g, MnSO4H2O 1g, cysteine salt
Hydrochlorate 0.1g, Tween 80 0.01ml, distilled water 800mL.
Lactobacillus solution: isolate and purify to obtain from Yoghourt or baby intestinal original strain (lactobacillus plantarum LP45, it is thermophilic
Lactobacillus lactis La28, bifidobacterium lactis BAL531, bifidobacterium bifidum TMC3115, Lactobacillus rhamnosus LR519, rhamnose cream
Bacillus LR863), original strain mixes after above-mentioned MRS liquid culture medium culture to logarithmic phase with 20% glycerol grade ratio
Uniformly, it is frozen in -80 DEG C up to seed liquor, lactobacillus solution is provided by Hebei Yiran Biological Technology Co., Ltd.'s strain library.
(1) fermentation liquid: taking lactobacillus solution to be inoculated into 10ml MRS liquid culture medium by 2% inoculum concentration, concussion
It mixes, 30 DEG C of culture 48h, successively continuously activates three generations, then be inoculated into 250ml modified MRS Liquid Culture by 10% inoculum concentration
Expand culture in base, wherein bifidobacterium lactis BAL531, bifidobacterium bifidum TMC3115 palpus Anaerobic culturel, other incubation steps
It is identical；
Cultured fermentation liquid takes 100ml to get fermentation liquid bulk samples, dilutes stoste with 0.9% sodium chloride solution, obtains
10 times of dilution samples, it is spare in 4 DEG C of refrigerations；
(2) inactivate fermentation liquid: 110 DEG C of heating 10min of fermentation liquid must inactivate fermentation liquid bulk samples, with 0.9% sodium chloride
Solution dilutes stoste, obtains 10 times of dilution samples；
(3) supernatant: fermentation liquid 7000r/min is centrifuged 20min and collects supernatant to get supernatant bulk samples, uses
0.9% sodium chloride solution dilutes stoste, obtains 10 times of dilution samples, spare in 4 DEG C of refrigerations；
(4) inactivate supernatant: 110 DEG C of heating 10min of supernatant must inactivate supernatant bulk samples, with 0.9% sodium chloride
Solution dilutes stoste, obtains 10 times of dilution samples；
(5) thallus: centrifugation gained bacterium mud is centrifuged after being sufficiently resuspended with 0.9% sodium chloride solution, in triplicate up to thallus
Bulk samples dilute stoste with 0.9% sodium chloride solution, obtain 10 times of dilution samples, and 100 times of dilution samples are refrigerated in 4 DEG C
It is spare；
(6) inactivate thallus: 130 DEG C of heating 10min of thallus bulk samples must inactivate thallus bulk samples, with 0.9% chlorination
Sodium solution dilutes stoste, obtains 10 times of dilution samples, 100 times of dilution samples；
(7) lysate: pasteurization is after thallus and protective agent are mixed and stirred for uniformly according to 1:2 to get lysate
Sample dilutes stoste with 0.9% sodium chloride solution, obtains 10 times of dilution samples, 100 times of dilution samples；
(8) dead bacterium: fermentation liquid is freeze-dried with 60 DEG C of sterilizing 4h after 9000r/min is centrifuged emulsification 15min up to dead bacterium
Sample is configured to 5x10 with 0.9% sodium chloride solution and dead bacterium respectively8CFU/ml、1x109CFU/ml、5x109CFU/ml is dense
The fluid sample of degree.
(9) culture of propionibacterium acnes: propionibacterium acnes seed liquor is taken to be inoculated into 10ml liquid by 3% inoculum concentration
Sulphur glycollate culture medium, concussion mix, 37 DEG C of Anaerobic culturel 48h, successively continuously spare after activation three generations.
2.2 processing
(1) it is cooled to 50 DEG C or so containing 2.0% element agar culture medium, 10mL is taken to be poured in sterilized petri dishes, cleaning work
It is dried in platform；
(2) aseptic nipper takes sterilizing Oxford cup to be placed in the plate of element agar, stands 10min；
(3) 1mL propionibacterium acnes 100mL is added to be cooled in 50 DEG C of thioglycollate medium and fill
Divide and mixes；
(4) topple over 10mL aforesaid liquid sulphur glycollate culture medium on the plate for put well Oxford cup, it is to be solidified after by ox
Saliva cup presss from both sides out, forms Oxford cup aperture；
(5) 0.1mL each sample is injected in each Oxford cup aperture, in 37 DEG C of standing Anaerobic culturel 72h.And with 0.9%
Sodium chloride solution is negative control；Fermentation liquid, fire extinguishing fermentation liquid, supernatant and inactivation supernatant are with 30 μ g/ml quadracyclines
For positive control, thallus, inactivation thallus, lysate and dead bacterium are using 34 μ g/ml quadracyclines as positive control.
2.3 post-processing
Table 2
2.4 analysis
With the diameter of vernier caliper measurement inhibition zone, measurement result such as table 1, wherein evaluation index are as follows: inhibition zone is straight
Diameter=0 does not have fungistatic effect then；Antibacterial circle diameter > 0, then have fungistatic effect；0 < antibacterial circle diameter < 18, antibacterial effect
Fruit is general；18≤antibacterial circle diameter < 27mm, fungistatic effect are good；Antibacterial circle diameter >=27mm, fungistatic effect are significant；"-" table
Show and does not do experiment.The selection result is as follows:
(1) the Oxford cup bacteriostatic test of fermentation liquid shows: the antibacterial circle diameter of LP45, TMC3115, LR519 and LR863 are big
In 30mm, fungistatic effect is significant, and the fungistatic effect of La28, BAL531 are good；By inactivation after LP45, La28, TMC3115,
LR519 and LR863 only has stoste to have fungistatic effect, and effect is general, and the inactivation fermentation liquid of BAL531 does not have fungistatic effect.
(2) the Oxford cup bacteriostatic test of supernatant shows: the supernatant of LP45, BAL531, TMC3115 and LR863 are antibacterial
Loop diameter about 30mm, fungistatic effect is significant, and the fungistatic effect of La28 is good, and only stoste has suppression after above-mentioned five kinds of bacterial strains inactivation
Bacterium effect, effect are general；LR519 only has stoste to have fungistatic effect, and effect is general.
(3) the Oxford cup bacteriostatic test of thallus shows: the fungistatic effect of LP45, TMC3115, LR519 and LR863 are significant,
BAL531 fungistatic effect is good；La28 only has stoste to have good fungistatic effect；Six kinds of bacterial strains do not have antibacterial after inactivation
Effect.
(3) the Oxford cup bacteriostatic test of lysate shows: the fungistatic effect of LP45, TMC3115 and LR863 are significant,
Middle LR863 lysate fungistatic effect is best, and LR519 fungistatic effect is good, and BAL531 only has stoste to have certain antibacterial effect
Fruit, La28 lysate is without fungistatic effect.
(4) the Oxford cup bacteriostatic test of dead bacterium shows: in LR519 and LR863, only LR863 has fungistatic effect, and antibacterial effect
Fruit increases with LR863 concentration and is increased, and fungistatic effect is good when wherein concentration is 5,000,000,000, and concentration is that 1,000,000,000 fungistatic effects are general,
Such as Fig. 8.
To sum up, the fermentation liquid of LP45, TMC3115 and LR863, supernatant, thallus and lysate are to propionibacterium acnes
Fungistatic effect is significant；And fermentation liquid, the thallus of LR519 are also very significant to propionibacterium acnes fungistatic effect；Also
The supernatant of BAL531 is significant to propionibacterium acnes fungistatic effect, and skin-protection product is used for after being further processed；
The dead bacterium of LR863 has fungistatic effect, and fungistatic effect is reduced with LR863 concentration and is deteriorated, can be straight as needed
Selecting takes suitable concentration for skin care item such as facial masks.
2.5 reference examples
Reference examples 1
Identical to treatment process as above-mentioned stock, difference, which is only that, replaces with Liquid Culture for MRS liquid culture medium
Base 1, according to the mass fraction, fluid nutrient medium 1 include 10 parts of glucose, 5 parts of peptones, 2 parts of sodium acetates and 83 parts of distilled water.
Reference examples 2
Identical to treatment process as above-mentioned stock, difference, which is only that, replaces with Liquid Culture for MRS liquid culture medium
Base 2, according to the mass fraction, fluid nutrient medium 2 include 15 parts of glucose, 8 parts of peptones, 4 parts of sodium acetates and 73 parts of distilled water.
Reference examples 3
Identical to last handling process as above-mentioned stock, difference, which is only that, replaces with liquid training for MRS liquid culture medium
Base 3 is supported, according to the mass fraction, fluid nutrient medium 3 includes 20 parts of glucose, 10 parts of peptones, 5 parts of sodium acetates and 65 parts of distillations
Water.
For culture medium described in above-mentioned reference examples 1-3 compared with MRS liquid culture medium, MRS liquid culture medium makes bacterium
Strain is growing more rapidly, and metabolite is more, and antibacterial circle diameter is bigger, more preferable to the inhibitory effect of propionibacterium acnes.
Embodiment 3
3.1 stock
Propionibacterium acnes, sulphur glycollate culture medium, 0.9% sodium chloride solution, 30 μ g/ml quadracycline,
The quadracycline of 34 μ g/ml is same as Example 1, and difference is only that MRS liquid culture medium and lactobacillus solution；
Protective agent: including 20 portions of skimmed milks, 6 portions of trehaloses, 2 parts of vitamin Cs, 3 parts of sodium glutamates, 60 parts of distilled water.
MRS liquid culture medium: including glucose 15g, peptone 8g, powdered beef 4g, yeast extract 3g, sodium acetate
4g, diammonium hydrogen citrate 1g, dipotassium hydrogen phosphate 1.5g, MgSO4·7H2O 0.5g, MnSO4H2O 0.6g, cysteine hydrochloric acid
Salt 0.3g, Tween 80 0.08ml, distilled water 900mL.
Lactobacillus solution: isolate and purify to obtain from Yoghourt or baby intestinal original strain (lactobacillus plantarum LP45, it is thermophilic
Lactobacillus lactis La28, bifidobacterium lactis BAL531, bifidobacterium bifidum TMC3115, Lactobacillus rhamnosus LR519, rhamnose cream
Bacillus LR863), original strain mixes after above-mentioned MRS liquid culture medium culture to logarithmic phase with 25% glycerol grade ratio
Uniformly, it is frozen in -70 DEG C up to seed liquor, lactobacillus solution is provided by Hebei Yiran Biological Technology Co., Ltd.'s strain library.
(1) fermentation liquid: taking lactobacillus solution to be inoculated into 10ml MRS liquid culture medium by 10% inoculum concentration, shake
Mixing is swung, 40 DEG C of culture 36h successively continuously activate three generations, then are inoculated into 250ml modified MRS Liquid Culture by 2% inoculum concentration
Expand culture in base, wherein bifidobacterium lactis BAL531, bifidobacterium bifidum TMC3115 palpus Anaerobic culturel, other incubation steps
It is identical；
Cultured fermentation liquid takes 100ml to get fermentation liquid bulk samples, dilutes stoste with 0.9% sodium chloride solution, obtains
10 times of dilution samples, it is spare in 4 DEG C of refrigerations；
(2) inactivate fermentation liquid: 130 DEG C of heating 20min of fermentation liquid must inactivate fermentation liquid bulk samples, with 0.9% sodium chloride
Solution dilutes stoste, obtains 10 times of dilution samples；
(3) supernatant: fermentation liquid 9000r/min is centrifuged 15min and collects supernatant to get supernatant bulk samples, uses
0.9% sodium chloride solution dilutes stoste, obtains 10 times of dilution samples, spare in 4 DEG C of refrigerations；
(4) inactivate supernatant: 130 DEG C of heating 20min of supernatant must inactivate supernatant bulk samples, with 0.9% sodium chloride
Solution dilutes stoste, obtains 10 times of dilution samples；
(5) thallus: centrifugation gained bacterium mud is centrifuged after being sufficiently resuspended with 0.9% sodium chloride solution, in triplicate up to thallus
Bulk samples dilute stoste with 0.9% sodium chloride solution, obtain 10 times of dilution samples, and 100 times of dilution samples are refrigerated in 4 DEG C
It is spare；
(6) inactivate thallus: 110 DEG C of heating 20min of thallus bulk samples must inactivate thallus bulk samples, with 0.9% chlorination
Sodium solution dilutes stoste, obtains 10 times of dilution samples, 100 times of dilution samples；
(7) lysate: pasteurization is after thallus and protective agent are mixed and stirred for uniformly according to 1:2 to get lysate
Sample dilutes stoste with 0.9% sodium chloride solution, obtains 10 times of dilution samples, 100 times of dilution samples；
(8) dead bacterium: fermentation liquid is freeze-dried with 80 DEG C of sterilizing 2h after 7000r/min is centrifuged emulsification 15min up to dead bacterium
Sample is configured to 5x10 with 0.9% sodium chloride solution and dead bacterium respectively8CFU/ml、1x109CFU/ml、5x109CFU/ml is dense
The fluid sample of degree.
(9) culture of propionibacterium acnes: propionibacterium acnes seed liquor is taken to be inoculated into 10ml liquid by 3% inoculum concentration
Sulphur glycollate culture medium, concussion mix, 37 DEG C of Anaerobic culturel 48h, successively continuously spare after activation three generations.
3.2 processing
(1) it is cooled to 50 DEG C or so containing 2.0% element agar culture medium, 10mL is taken to be poured in sterilized petri dishes, cleaning work
It is dried in platform；
(2) aseptic nipper takes sterilizing Oxford cup to be placed in the plate of element agar, stands 10min；
(3) 1mL propionibacterium acnes 100mL is added to be cooled in 50 DEG C of thioglycollate medium and fill
Divide and mixes；
(4) topple over 10mL aforesaid liquid sulphur glycollate culture medium on the plate for put well Oxford cup, it is to be solidified after by ox
Saliva cup presss from both sides out, forms Oxford cup aperture；
(5) 0.1mL each sample is injected in each Oxford cup aperture, in 37 DEG C of standing Anaerobic culturel 60h.And with 0.9%
Sodium chloride solution is negative control；Fermentation liquid, fire extinguishing fermentation liquid, supernatant and inactivation supernatant are with 30 μ g/ml quadracyclines
For positive control, thallus, inactivation thallus, lysate and dead bacterium are using 34 μ g/ml quadracyclines as positive control.
3.3 post-processing
Table 3
3.4 analysis
With the diameter of vernier caliper measurement inhibition zone, measurement result such as table 1, wherein evaluation index are as follows: inhibition zone is straight
Diameter=0 does not have fungistatic effect then；Antibacterial circle diameter > 0, then have fungistatic effect；0 < antibacterial circle diameter < 18, antibacterial effect
Fruit is general；18≤antibacterial circle diameter < 27mm, fungistatic effect are good；Antibacterial circle diameter >=27mm, fungistatic effect are significant；"-" table
Show and does not do experiment.The selection result is as follows:
(1) the Oxford cup bacteriostatic test of fermentation liquid shows: the antibacterial circle diameter of LP45, TMC3115, LR519 and LR863 are big
In 30mm, fungistatic effect is significant, and the fungistatic effect of La28, BAL531 are good；By inactivation after LP45, La28, TMC3115,
LR519 and LR863 only has stoste to have fungistatic effect, and effect is general, and the inactivation fermentation liquid of BAL531 does not have fungistatic effect.
(2) the Oxford cup bacteriostatic test of supernatant shows: the supernatant of LP45, BAL531, TMC3115 and LR863 are antibacterial
Loop diameter about 30mm, fungistatic effect is significant, and the fungistatic effect of La28 is good, and only stoste has suppression after above-mentioned five kinds of bacterial strains inactivation
Bacterium effect, effect are general；LR519 only has stoste to have fungistatic effect, and effect is general.
(3) the Oxford cup bacteriostatic test of thallus shows: the antibacterial effect of LP45, BAL531, TMC3115, LR519 and LR863
Fruit is significant, and La28 only has stoste to have good fungistatic effect；Six kinds of bacterial strains do not have fungistatic effect after inactivation.
(3) the Oxford cup bacteriostatic test of lysate shows: the fungistatic effect of LP45, TMC3115, LR519 and LR863 are aobvious
It writes, wherein LR863 lysate fungistatic effect is best, and BAL531 only has stoste to have certain fungistatic effect, and La28 lysis produces
Object is without fungistatic effect.
(4) the Oxford cup bacteriostatic test of dead bacterium shows: in LR519 and LR863, only LR863 has fungistatic effect, and antibacterial effect
Fruit increases with LR863 concentration and is increased, and fungistatic effect is good when wherein concentration is 5,000,000,000, and 1,000,000,000 antibacterial circle diameters are 17.24mm,
Fungistatic effect is general.
To sum up, the fermentation liquid of LP45, TMC3115 and LR863, supernatant, thallus and lysate are to propionibacterium acnes
Fungistatic effect is significant；And fermentation liquid, thallus and the lysate of LR519 are also very aobvious to propionibacterium acnes fungistatic effect
It writes；There are also the supernatant of BAL531 and thallus are significant to propionibacterium acnes fungistatic effect, skin care is used for after being further processed
Product；
The dead bacterium of LR863 has fungistatic effect, and fungistatic effect is reduced with LR863 concentration and is deteriorated, can be straight as needed
Selecting takes suitable concentration for skin care item such as facial masks.
3.5 reference examples
Reference examples 1
Identical to treatment process as above-mentioned stock, difference, which is only that, replaces with Liquid Culture for MRS liquid culture medium
Base 1, according to the mass fraction, fluid nutrient medium 1 include 10 parts of glucose, 5 parts of peptones, 2 parts of sodium acetates and 83 parts of distilled water.
Reference examples 2
Identical to treatment process as above-mentioned stock, difference, which is only that, replaces with Liquid Culture for MRS liquid culture medium
Base 2, according to the mass fraction, fluid nutrient medium 2 include 15 parts of glucose, 8 parts of peptones, 4 parts of sodium acetates and 73 parts of distilled water.
Reference examples 3
Identical to last handling process as above-mentioned stock, difference, which is only that, replaces with liquid training for MRS liquid culture medium
Base 3 is supported, according to the mass fraction, fluid nutrient medium 3 includes 20 parts of glucose, 10 parts of peptones, 5 parts of sodium acetates and 65 parts of distillations
Water.
For culture medium described in above-mentioned reference examples 1-3 compared with MRS liquid culture medium, MRS liquid culture medium makes bacterium
Strain is growing more rapidly, and metabolite is more, and antibacterial circle diameter is bigger, more preferable to the inhibitory effect of propionibacterium acnes.
Two, Application Example
Example 1
According to the mass fraction, dead bacterium of the facial mask essence by 0.5 part of Lactobacillus rhamnosus LR863,10 parts of glycerol, 0.05 part
With being combined into, Deacne pack carrier chooses non-woven fabrics for Sodium Hyaluronate and 89.45 parts of distilled water；
Wherein to the dead bacterium filtration sterilization of Lactobacillus rhamnosus LR863, make bacterial population 900CFU/ml.
Example 2
According to the mass fraction, thallus of the facial mask essence by 0.3 part of lactobacillus plantarum LP45,0.3 part of Lactobacillus rhamnosus
The lysate of LR519,13 parts of glycerol, 0.08 part of Sodium Hyaluronate and 86.32 parts of distilled water, which are matched, to be combined into, Deacne pack carrier
Choose silk；
Wherein to the thallus filtration sterilization of lactobacillus plantarum LP45, make bacterial population 500CFU/ml, to Lactobacillus rhamnosus
The lysate filtration degerming of LR519, makes bacterial population 400CFU/ml.
Example 3
According to the mass fraction, thallus of the facial mask essence by 0.3 part of bifidobacterium lactis BAL531,11 parts of glycerol, 0.04 part
With being combined into, Deacne pack carrier chooses biological fiber and tencel fiber for Sodium Hyaluronate and 88.66 parts of distilled water；
Wherein to the thallus filtration sterilization of bifidobacterium lactis BAL531, make bacterial population 800CFU/ml.
Example 4
According to the mass fraction, acne eliminating cream includes the dead bacterium of 0.5 part of Lactobacillus rhamnosus LR863,4.8 parts of vaseline, and 3.84
Part octadecyl alcolol, 1.44 parts of monoglycerides, 2.88 parts of Valelinum Liquidums, 0.48 part of lauryl sodium sulfate, 2.4 parts of glycerol, 83.66 parts of distillations
Water；
Wherein to the dead bacterium filtration sterilization of Lactobacillus rhamnosus LR863, make bacterial population 700CFU/ml.
Example 5
According to the mass fraction, acne eliminating cream includes the thallus of 0.4 part of bifidobacterium bifidum TMC3115,5.2 parts of vaseline,
3.34 parts of octadecyl alcolols, 1.5 parts of monoglycerides, 2.72 parts of Valelinum Liquidums, 0.31 part of lauryl sodium sulfate, 3.53 parts of glycerol, 83 parts of steamings
Distilled water；
Wherein to the thallus filtration sterilization of bifidobacterium bifidum TMC3115, make bacterial population 600CFU/ml.
Example 6
According to the mass fraction, acne eliminating cream includes the fermentation liquid of 0.2 part of bifidobacterium bifidum TMC3115,0.5 part of rhamnose cream
The thallus of bacillus LR863,5.5 parts of vaseline, 3.3 parts of octadecyl alcolols, 2.5 parts of monoglycerides, 2.98 parts of Valelinum Liquidums, 0.54 part of dodecane
Base sodium sulphate, 2.8 parts of glycerol, 81.68 parts of distilled water；
Desalting processing wherein is inactivated to the fermentation liquid of bifidobacterium bifidum TMC3115, makes bacterial population 300CFU/ml, it is right
The thallus filtration sterilization of Lactobacillus rhamnosus LR863, makes bacterial population 500CFU/ml.
Example 7
According to the mass fraction, the supernatant of lactobacillus acidophilus La28 takes 5 parts, and the thallus of bifidobacterium lactis BAL531 takes 5 parts,
With 10 parts of glycerol, 0.08 part of Sodium Hyaluronate, 0.06 part of niacinamide, 1 part of triglycerides, 0.07 part of EDETATE SODIUM, which is matched, is combined into shield
Skin Essence；
Wherein the supernatant of lactobacillus acidophilus La28 inactivates desalting processing, makes bacterial population 500CFU/ml, bifidobacterium lactis
The thallus filtration sterilization of BAL531, makes bacterial population 200CFU/ml.
Example 8
According to the mass fraction, 9 portions of Lactobacillus rhamnosus LR863 supernatants and 11.5 parts of glycerol, 0.05 part of hyaluronic acid are taken
Sodium, 0.05 part of niacinamide, 2 parts of triglycerides, 0.05 part of EDETATE SODIUM, which is matched, is combined into facial treatment essence liquid；
Wherein Lactobacillus rhamnosus LR863 supernatant inactivates desalting processing, makes bacterial population 850CFU/ml.
Example 9
According to the mass fraction, the thallus and 13.4 parts of glycerol of 12 parts of Lactobacillus rhamnosus LR863,0.09 part of hyaluronic acid
Sodium, 0.04 part of niacinamide, 1.5 parts of triglycerides, 0.08 part of EDETATE SODIUM, which is matched, is combined into facial treatment essence liquid；
The wherein thallus filtration sterilization of Lactobacillus rhamnosus LR863, makes bacterial population 350CFU/ml.
Claims (10)
1. lactic acid bacteria inhibit propionibacterium acnes in application, which is characterized in that the lactic acid bacteria be lactic acid bacteria thallus and/or
Lactic acid bacteria derivative；
Lactic acid bacteria thallus includes lactobacillus plantarum (Lactobacillus plantarum) LP45, CGMCC No.8072, acidophilus
Lactobacillus (Lactobacillus acidophilus) La28, CGMCC No.11506, bifidobacterium lactis
(Bifidobacteriumlactis) BAL531, CGMCC No.17329, bifidobacterium bifidum
(Bifidobacteriumbifidum) TMC3115, CGMCC No.8462, Lactobacillus rhamnosus (Lactobacillus
Rhamnosus) LR519, CGMCC No.15969, Lactobacillus rhamnosus (Lactobacillus rhamnosus) LR863,
One or more of CGMCC No.14410；
The lactic acid bacteria derivative includes the inactivation fermentation liquid, the supernatant of LP45, LP45 of the fermentation liquid of above-mentioned LP45, LP45
Inactivation supernatant, the lysate of LP45, the fermentation liquid of La28, the inactivation fermentation liquid of La28, the supernatant of La28, La28 go out
Living supernatant, the fermentation liquid of BAL531, the supernatant of BAL531, the inactivation supernatant of BAL531, BAL531 lysate,
The fermentation liquid of TMC3115, the inactivation fermentation liquid of TMC3115, the supernatant of TMC3115, TMC3115 inactivation supernatant,
The lysis production of the lysate of TMC3115, the fermentation liquid of LR519, the inactivation fermentation liquid of LR519, the supernatant of LR519, LR519
Object, the fermentation liquid of LR863, LR863 inactivation fermentation liquid, the supernatant of LR863, LR863 inactivation supernatant, LR863 it is molten
One or more of born of the same parents' product or the dead bacterium of LR863.
2. lactic acid bacteria as described in claim 1 is inhibiting the application in propionibacterium acnes, which is characterized in that the hair of the LP45
Zymotic fluid, the fermentation liquid of La28, the fermentation liquid of BAL531, the fermentation liquid of TMC3115, the fermentation liquid of LR519 or LR863 fermentation liquid
Prepared by following steps: LP45, La28, BAL531, TMC3115, LR519 or LR863 are by fluid nutrient medium culture to logarithmic phase
It is uniformly mixed afterwards with the glycerol of 20%-30% grade ratio, freezes to obtain seed liquor in -60 DEG C~-80 DEG C；
Take seed liquor to be inoculated into 10ml fluid nutrient medium by the inoculum concentration of 2%-10%, concussion mixes, 30 DEG C of -40 DEG C of cultures for 24 hours -
48h, successively continuously activation three generations, then by the inoculum concentration of 2%-10% be inoculated into 250ml fluid nutrient medium expand culture to get
The fermentation liquid of LP45, the fermentation liquid of La28, the fermentation liquid of BAL531, the fermentation liquid of TMC3115, LR519 fermentation liquid or LR863
Fermentation liquid；
The fermentation liquid of the fermentation liquid or TMC3115 of above-mentioned BAL531 is prepared as anaerobic condition；
The fermentation liquid of LP45, the fermentation liquid of La28, the fermentation liquid of TMC3115, the fermentation liquid of LR519 or the fermentation liquid of LR863 exist
110-130 DEG C of heating 10-20min ferments to get the inactivation of the inactivation fermentation liquid of LP45, the inactivation fermentation liquid, TMC3115 of La28
Liquid, the inactivation fermentation liquid of LR519 or the inactivation fermentation liquid of LR863.
3. lactic acid bacteria as claimed in claim 2 is inhibiting the application in propionibacterium acnes, which is characterized in that the hair of the LP45
Zymotic fluid, the fermentation liquid of La28, the fermentation liquid of BAL531, the fermentation liquid of TMC3115, the fermentation liquid of LR519 or LR863 fermentation liquid
After 7000-9000r/min is centrifuged 10-15min, supernatant is collected to get the supernatant of LP45, the supernatant of La28, BAL531
Supernatant, the supernatant of TMC3115, the supernatant of LR519 or LR863 supernatant；
The supernatant of LP45, the supernatant of La28, the supernatant of BAL531, the supernatant of TMC3115 or the supernatant of LR863 exist
110-130 DEG C of heating 10-20min is to get the inactivation supernatant of LP45, the inactivation supernatant for inactivating supernatant, TMC3115 of La28
The inactivation supernatant of liquid or LR863；
The fermentation of the fermentation liquid of the LP45, the fermentation liquid of La28, the fermentation liquid of BAL531, the fermentation liquid of TMC3115, LR519
The fermentation liquid of liquid or LR863 centrifugation gained bacterium mud is sufficiently resuspended with 0.9% sodium chloride solution after, repeatedly 7000-9000r/min from
Heart 10-15min is three times to get the bacterium of the thallus of LP45, the thallus of La28, the thallus of BAL531, the thallus of TMC3115, LR519
The thallus of body or LR863；
The fermentation liquid of the LR863 is freeze-dried after 7000-9000r/min is centrifuged 10-15min with 60-80 DEG C of sterilizing 2-4h,
Up to the dead bacterium of LR863.
4. lactic acid bacteria as claimed in claim 2 is inhibiting the application in propionibacterium acnes, which is characterized in that the bacterium of the LP45
Body, the thallus of BAL531, the thallus of TMC3115, the thallus of LR519 or LR863 thallus mix and stir according to 1:2 with protective agent
Mix uniformly after pasteurization to get the lysate of LP45, the lysate of BAL531, TMC3115 lysate, LR519
Lysate or LR863 lysate.
5. lactic acid bacteria as claimed in claim 4 is inhibiting the application in propionibacterium acnes, which is characterized in that press mass fraction
Meter, the protective agent includes 10-15 portions of skimmed milks, 2-6 portions of trehaloses, 1.5-2 parts of vitamin Cs, 1-3 parts of sodium glutamates, 60-80
Part distilled water.
6. lactic acid bacteria as claimed in claim 2 is inhibiting the application in propionibacterium acnes, which is characterized in that press mass fraction
Meter, the fluid nutrient medium includes 10-20 parts of glucose, 5-10 parts of peptones, 2-5 parts of sodium acetates and 65-83 parts of distilled water.
7. lactic acid bacteria as claimed in claim 2 is inhibiting the application in propionibacterium acnes, which is characterized in that press mass fraction
Meter, the fluid nutrient medium include 10-20 parts of glucose, 5-10 parts of peptones, 2-6.5 parts of powdered beefs, 1-5 parts of yeast extracts,
2-5 parts of sodium acetates, 0.5-2 parts of diammonium hydrogen citrates, 0.8-2 parts of dipotassium hydrogen phosphates, 0.32-0.58 parts of MgSO4·7H2O, 0.25-
1 part of MnSO4H2O, 0.1-0.5 parts of cysteine hydrochlorides, 0.01-0.1 parts of Tween 80s, 800-1000 parts of distilled water.
8. it is a kind of using lactic acid bacteria as described in claim 1 inhibit in propionibacterium acnes using manufactured Deacne pack,
Deacne pack is made of facial mask essence and face-mask material, which is characterized in that according to the mass fraction, facial mask essence includes 0.3
- 0.6 part of lactic acid bacteria thallus of part and/or lactic acid bacteria derivative, 10-13 parts of glycerol, 0.04-0.08 parts of Sodium Hyaluronates and 86.32-
89.45 parts of distilled water, face-mask material include one or more of non-woven fabrics, silk, biological fiber or tencel fiber.
9. it is a kind of using probiotics as described in claim 1 inhibit in propionibacterium acnes using manufactured acne eliminating cream,
It is characterized in that, according to the mass fraction, acne eliminating cream includes 0.4-0.7 parts of probio thallines and/or probiotic derived object, 4.8-5.5
Part vaseline, 3.3-3.84 parts of octadecyl alcolols, 1.44-2.5 parts of monoglycerides, 2.72-2.98 parts of Valelinum Liquidums, 0.31-0.54 part 12
Sodium alkyl sulfate, 2.4-3.53 parts of glycerol, 81.68-83 parts of distilled water.
10. it is a kind of using probiotics as described in claim 1 inhibit propionibacterium acnes in using manufactured facial treatment essence
Liquid, which is characterized in that according to the mass fraction, including 9-12 parts of probio thallines and/or probiotic derived object, 10-13.4 parts are sweet
Oil, 0.05-0.09 parts of Sodium Hyaluronates, 0.04-0.06 parts of niacinamide, 1-2 parts of triglycerides, 0.05-0.08 parts of EDETATE SODIUMs.
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| CN110982741A (en) * | 2019-12-11 | 2020-04-10 | 河北一然生物科技有限公司 | Preparation method and application of lactobacillus with tyrosinase activity inhibiting function |
| CN111202709A (en) * | 2019-12-11 | 2020-05-29 | 河北一然生物科技有限公司 | Lactobacillus facial mask capable of inhibiting propionibacterium acnes and preparation method thereof |
| CN111904923A (en) * | 2020-08-11 | 2020-11-10 | 江苏华能药业有限公司 | Probiotic and metazoan composition set |
| CN112057479A (en) * | 2020-11-16 | 2020-12-11 | 招远新兴化工有限公司 | Probiotic composition capable of relieving diarrhea caused by antibiotics |
| CN112094790A (en) * | 2020-11-17 | 2020-12-18 | 招远新兴化工有限公司 | Lactobacillus plantarum LP45 live bacterium preparation for regulating intestinal flora and application thereof |
| CN113576943A (en) * | 2021-07-09 | 2021-11-02 | 中山大学 | Prebiotics fermentation product and preparation method and application thereof |
| CN114058547A (en) * | 2021-11-29 | 2022-02-18 | 中山大学 | Lactobacillus fermentum, fermentation broth of lactobacillus fermentum and application of fermentation broth |
| CN114317617A (en) * | 2021-12-20 | 2022-04-12 | 无锡弘焕微生态科技有限公司 | Preparation method and application of triple probiotic fermented compound with anti-wrinkle effect |
| CN114681588A (en) * | 2022-02-17 | 2022-07-01 | 中山大学 | Application of polypeptide EN-9 in preparation of product for treating acne |
| CN114869839A (en) * | 2022-06-06 | 2022-08-09 | 绽妍生物科技有限公司 | Multilayer microencapsulated mild composite acid repair gel based on lactic acid bacteria fermentation product EPS and preparation method and application thereof |
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| CN110982741A (en) * | 2019-12-11 | 2020-04-10 | 河北一然生物科技有限公司 | Preparation method and application of lactobacillus with tyrosinase activity inhibiting function |
| CN111202709A (en) * | 2019-12-11 | 2020-05-29 | 河北一然生物科技有限公司 | Lactobacillus facial mask capable of inhibiting propionibacterium acnes and preparation method thereof |
| CN111904923A (en) * | 2020-08-11 | 2020-11-10 | 江苏华能药业有限公司 | Probiotic and metazoan composition set |
| CN112057479A (en) * | 2020-11-16 | 2020-12-11 | 招远新兴化工有限公司 | Probiotic composition capable of relieving diarrhea caused by antibiotics |
| CN112094790A (en) * | 2020-11-17 | 2020-12-18 | 招远新兴化工有限公司 | Lactobacillus plantarum LP45 live bacterium preparation for regulating intestinal flora and application thereof |
| CN112094790B (en) * | 2020-11-17 | 2021-03-02 | 河北一然生物科技有限公司 | Lactobacillus plantarum LP45 live bacterium preparation for regulating intestinal flora and application thereof |
| CN113576943A (en) * | 2021-07-09 | 2021-11-02 | 中山大学 | Prebiotics fermentation product and preparation method and application thereof |
| CN113576943B (en) * | 2021-07-09 | 2022-07-12 | 中山大学 | A kind of prebiotic fermentation product and its preparation method and application |
| CN114058547A (en) * | 2021-11-29 | 2022-02-18 | 中山大学 | Lactobacillus fermentum, fermentation broth of lactobacillus fermentum and application of fermentation broth |
| CN114058547B (en) * | 2021-11-29 | 2023-12-05 | 广州集妍化妆品科技有限公司 | Lactobacillus fermentum, lactobacillus fermentum fermentation liquid and application thereof |
| CN114317617A (en) * | 2021-12-20 | 2022-04-12 | 无锡弘焕微生态科技有限公司 | Preparation method and application of triple probiotic fermented compound with anti-wrinkle effect |
| CN114681588A (en) * | 2022-02-17 | 2022-07-01 | 中山大学 | Application of polypeptide EN-9 in preparation of product for treating acne |
| CN114681588B (en) * | 2022-02-17 | 2023-06-06 | 中山大学 | Application of polypeptide EN-9 in preparation of products for treating acne |
| CN114869839A (en) * | 2022-06-06 | 2022-08-09 | 绽妍生物科技有限公司 | Multilayer microencapsulated mild composite acid repair gel based on lactic acid bacteria fermentation product EPS and preparation method and application thereof |
| CN114869839B (en) * | 2022-06-06 | 2023-10-20 | 绽妍生物科技有限公司 | Multilayer microencapsulated mild composite acid repair gel based on lactobacillus fermentation product EPS, and preparation method and application thereof |
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