CN116869870B - Galactose yeast-like fermentation product filtrate with moisturizing, tightening and relieving effects, and preparation method and application thereof - Google Patents
Galactose yeast-like fermentation product filtrate with moisturizing, tightening and relieving effects, and preparation method and application thereof Download PDFInfo
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- CN116869870B CN116869870B CN202310535346.0A CN202310535346A CN116869870B CN 116869870 B CN116869870 B CN 116869870B CN 202310535346 A CN202310535346 A CN 202310535346A CN 116869870 B CN116869870 B CN 116869870B
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Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
- A61K8/9728—Fungi, e.g. yeasts
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
- C12N1/165—Yeast isolates
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P1/00—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes
- C12P1/02—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes by using fungi
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/85—Products or compounds obtained by fermentation, e.g. yoghurt, beer, wine
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/645—Fungi ; Processes using fungi
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- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
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- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- General Health & Medical Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Animal Behavior & Ethology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Botany (AREA)
- Birds (AREA)
- Dermatology (AREA)
- Epidemiology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- General Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- Virology (AREA)
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Abstract
The invention relates to the technical field of microorganisms, and particularly discloses a galactose yeast-like fermentation product filtrate with moisturizing, tightening and relieving effects, and a preparation method and application thereof. The invention discovers by accident that the galactose yeast-like fermentation product filtrate prepared by fermenting the autonomously separated strain has good moisturizing, tightening and relieving effects, is safe and non-irritating, has simple preparation process, can realize industrial production, and is suitable for being applied to cosmetics.
Description
Technical Field
The invention relates to the technical field of microorganisms, in particular to a galactose yeast-like fermentation product filtrate with moisturizing, tightening and relieving effects, and a preparation method and application thereof.
Background
The galactose yeast-like fermentation product (raw material name: pitera) was obtained by fermentation from a strain of Saccharomyces Trichosporon Kashiwayama by Japanese scholars Kashiwayama. At present, the galactose yeast-like fermentation product filtrate has become one of the raw materials newly added in the catalogue of used cosmetic raw materials (2021 edition) published by the national drug administration. The galactose yeast-like fermentation product filtrate is rich in free amino acids, minerals, organic acids, inorganic acids, etc., and has various skin care effects.
At present, intensive research is carried out on a preparation method of the galactose yeast-like fermentation product filtrate, and the galactose yeast-like fermentation product filtrate which has the same good efficacy as Pitera is hoped to be prepared, but the galactose yeast-like fermentation product filtrate prepared by the existing method has poor moisturizing, tightening and relieving effects, and cannot meet the demands of consumers.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provide a galactose yeast-like fermentation product filtrate with moisturizing, tightening and relieving effects, and a preparation method and application thereof. The invention obtains the candida bailii through autonomous separation and screening, and the galactose yeast-like fermentation product filtrate obtained by fermenting the candida bailii contains more active ingredients, so that the candida bailii can be used in the field of cosmetics.
In order to achieve the above purpose, the technical scheme adopted by the invention is as follows:
the first object of the present invention is to provide a method for preparing a galactose yeast-like fermentation product filtrate having excellent moisturizing, tightening and soothing effects, comprising the steps of:
s1, preparing fermented grains: uniformly mixing sorghum, corn, rice, glutinous rice, wheat and water to obtain a mixture, steaming, cooling, adding distiller's yeast for saccharification and fermentation, and filtering to obtain fermented grains;
S2, preparing seed liquid: inoculating bacterial colonies of the candida berosa to a liquid culture medium for culture to obtain seed liquid;
s3, fermentation treatment: uniformly mixing the fermented grains obtained in the step S1, lactose, milk powder and water, and sterilizing to obtain a fermentation substrate; inoculating the seed liquid in the step S2 into the fermentation substrate for fermentation to obtain fermentation liquid;
s4, post-processing: and (3) carrying out high-pressure homogenization treatment, filtration treatment and sterilization treatment on the fermentation broth in the step (S4) to obtain galactose yeast-like fermentation product filtrate with moisturizing, tightening and relieving effects.
The Mortierella bainieri is preserved in China Center for Type Culture Collection (CCTCC) No. M2023647 at the preservation number of 2023, 4 and 27 days, and the preservation address is Wuhan in China.
The product obtained by fermenting the candida baileyi has more active ingredients, and can improve the efficacy of cosmetics.
The preparation method of the galactose yeast-like fermentation product filtrate is simple, and can realize industrial production. The prepared grain fermented grains have important influence on the fermentation process of the self-separated candida baileyi, the grain fermented grains prepared from the sorghum with the shells, the rice, the glutinous rice and the wheat are more beneficial to the fermentation of the candida baileyi, the prepared galactose yeast-like fermentation product filtrate is rich in various nutrients, the moisturizing, pore tightening, relieving and repairing effects of the galactose yeast-like fermentation product filtrate are improved, and the prepared galactose yeast-like fermentation product filtrate is safe and non-irritating.
The invention discloses a preparation method of a bacterial suspension by separating Brevibacterium from fermented grains of white spirit, and specifically preparing a sample of the fermented grains of the white spirit into a bacterial suspension; diluting the bacterial suspension, then coating the bacterial suspension in a Bengalia red culture medium, and culturing to obtain a colony with saccharomycetes characteristics; placing the bacterial colony in YPD solid culture medium for separation and purification, picking single bacterial colony with typical yeast bacterial colony characteristics, and placing the single bacterial colony in YPD solid culture medium for secondary separation and purification to obtain saccharomycetes; the saccharomycete is identified and then is determined to be the candida bailii.
The fermented grains of the white spirit contain a large amount of microorganisms, and different culture mediums are selected for separation and purification, so that the types of the obtained microorganisms are greatly different. According to the invention, the Bengalia amara culture medium and the YPD solid culture medium are selected to separate and purify microorganisms in the distilled grain of the white spirit, and the Brevibacterium having typical yeast colonies is selected to be used for preparing the galactose yeast-like fermentation product filtrate, so that the result shows that the prepared galactose yeast-like fermentation product filtrate has good moisturizing, pore tightening, relieving and repairing effects.
Meanwhile, in the fermentation matrix, the added grain, lactose and milk powder have synergistic effect, the candida bailii is fermented by the grain, lactose and milk powder, and the prepared galactose yeast-like fermentation product filtrate has better effects of moisturizing, pore tightening, relieving and repairing.
In the step S1, the mixture comprises 3-8% of sorghum, 9-13% of corn, 12-18% of rice, 2-5% of glutinous rice, 3-8% of wheat and the balance of water in percentage by mass; preferably, the mixture comprises 5% sorghum, 12% corn, 15% rice, 3% glutinous rice, 5% wheat, and the balance water.
According to the invention, sorghum, corn, rice, glutinous rice and wheat with the mass percentages are selected, so that the effects of moisture preservation, compactness, relaxation and the like of the galactose yeast-like fermentation product filtrate can be effectively improved.
As a preferred embodiment of the preparation method of the galactose yeast-like fermentation product filtrate, sorghum, rice, glutinous rice and wheat are all provided with shells. The sorghum, rice, glutinous rice and wheat with shells can improve the performance of the prepared galactose yeast-like fermentation product filtrate.
In some embodiments, in step S1, the distiller' S yeast is added in an amount of 0.2-1% of the mass of the mixture, preferably 0.5% of the mass of the mixture.
As a preferred embodiment of the preparation method of the galactose yeast-like fermentation product filtrate, in the step S1, the temperature of the saccharification and fermentation is 32-38 ℃, the time of the saccharification and fermentation is 24-38 hours, and the relative humidity of the saccharification and fermentation is 60-80%; preferably, the temperature of the saccharification and fermentation is 35 ℃, the time of the saccharification and fermentation is 36 hours, and the relative humidity of the saccharification and fermentation is 70%.
In some embodiments, in step S1, the temperature of the cooking is 90-105 ℃, and the time of the cooking is 20-60 min; preferably, the temperature of the cooking is 100 ℃, and the time of the cooking is 40min.
In some embodiments, in step S1, the filtering is mesh filtration or membrane filtration.
In some embodiments, in step S2, the liquid medium comprises at least one of PDB medium, potato integrated medium, wort medium, YM medium, schia medium.
More preferably, in step S2, the liquid medium is a potato integrated medium.
In some embodiments, in step S2, the temperature of the cultivation is 25-32 ℃, the time of the cultivation is 36-60 hours, and the rotation speed of the cultivation is 100-200 rpm; preferably, the temperature of the cultivation is 28 ℃, the time of the cultivation is 48 hours, and the rotation speed of the cultivation is 150rpm.
In the step S3, the fermentation substrate comprises 12-18% of grain, 1-2% of lactose, 3-8% of milk powder and the balance of water in percentage by mass; preferably, the fermentation substrate comprises 15% of grain, 1.5% of lactose, 5% of milk powder and the balance of water.
In some specific embodiments, in the step S3, the temperature of the sterilization treatment is 110-125 ℃, and the time of the sterilization treatment is 15-30 min; more preferably, the temperature of the sterilization treatment is 121 ℃, and the time of the sterilization treatment is 20min.
In some embodiments, in step S3, the seed solution is inoculated in an amount of 0.5 to 2%; preferably, the seed solution is inoculated in an amount of 1%.
In some embodiments, in step S3, noFermenting bacteria air with the ventilation amount of 30-60 dm 3 /(h.L); preferably, the sterile air is introduced in an amount of 50dm 3 /(h·L)。
In some embodiments, in step S3, the fermentation temperature is 25-32 ℃, the fermentation time is 18-36 hours, and the fermentation rotating speed is 120-220 rpm; preferably, the temperature of the fermentation is 28 ℃, the time of the fermentation is 24 hours, and the rotating speed of the fermentation is 180rpm.
In the step S4, the fermentation liquor is homogenized for 20-60 min under 8-12 MPa at high pressure; preferably, the fermentation broth is homogenized for 30min at a high pressure of 10 MPa; the sterilization treatment is high-pressure sterilization treatment, wherein the high-pressure sterilization treatment is to subject the filtered fermentation liquor to high-pressure sterilization under 300-800 MPa for 5-20 min; preferably, the autoclaving is performed by autoclaving the filtered broth at 500MPa for 10min.
The invention selects and places the filtered fermentation liquor at 300-800 MPa for high-pressure sterilization for 5-20 min, thereby avoiding the situation that the traditional pasteurization needs to sterilize at 80-105 ℃, the high-temperature condition easily causes the loss of active substances in the galactose yeast-like fermentation product filtrate, and the galactose yeast-like fermentation product filtrate has poorer moisturizing, repairing and the like effects.
In some embodiments, in step S4, the filtering treatment is to filter the high-pressure homogenized fermentation broth with a diatomite filter or with a filter membrane.
The second purpose of the invention is to provide a galactose yeast-like fermentation product filtrate prepared by the preparation method.
The galactose yeast-like fermentation product filtrate provided by the invention is fermented by adopting yeast, and has good moisturizing, tightening and relieving effects.
The third purpose of the invention is to provide the application of the galactose yeast-like fermentation product filtrate with moisturizing, tightening and relieving effects in preparing cosmetics.
In some embodiments, the galactose yeast-like fermentation product filtrate with moisturizing, tightening and soothing effects is added to the cosmetic in an amount of 0.1 to 100wt%.
Compared with the prior art, the invention has the following beneficial effects:
the invention provides a galactose yeast-like fermentation product filtrate with moisturizing, tightening and relieving effects, and a preparation method and application thereof. The galactose yeast-like fermentation product filtrate provided by the invention has good moisturizing, tightening and relieving effects, and is suitable for being applied to cosmetics.
Drawings
FIG. 1 is a morphology of isolated Candida bailii according to the present invention;
FIG. 2 is an optical microscopic image of isolated Brevibacterium flavum according to the present invention.
Detailed Description
For a better description of the objects, technical solutions and advantages of the present invention, the present invention will be further described with reference to the accompanying drawings and specific embodiments.
In the following examples and comparative examples, the experimental methods used were conventional methods unless otherwise specified, and the materials, reagents and the like used, unless otherwise specified, were all commercially available. The percentages mentioned in the examples and comparative examples are percentages by mass unless otherwise specified.
The unit of the access quantity of sterile air mentioned in the present invention is dm 3 /(h.L), where dm 3 Refers to the amount of sterile air introduced, h refers to the hours, and L refers to the volume of the fermentation substrate. The access to sterile air as mentioned in the present invention is 50dm 3 /(h.L), i.e., 50dm of sterile air was introduced per liter of fermentation substrate per hour 3 。
The Bengalia red culture medium, the YPD solid culture medium, the YPD liquid culture medium, the PDB culture medium, the potato comprehensive culture medium, the wort culture medium, the YM culture medium and the Chlamydia culture medium which are used in the invention are all culture media commonly used in the prior art, and the preparation method is as follows:
bengalbanum medium (1L Bengalbanum medium was prepared as an example): peptone 5g, glucose 10g, KH 2 PO 4 1g、MgSO 4 ·7H 2 100mL of 1/3000 Bengal solution containing 0.5g of O and 0.1g of chloramphenicol, distilled water was added to 1L, and the mixture was sterilized at 121℃for 20 minutes and stored for use.
YPD solid medium (example of YPD solid medium prepared 1L): 10g of yeast extract powder, 15g of peptone, 15g of glucose and 20g of agar powder, adding distilled water to 1L, sterilizing at 121 ℃ for 20min, and storing for later use.
YPD liquid medium (example of YPD liquid medium prepared in 1L): 10g of yeast extract powder, 15g of peptone and 15g of glucose, adding distilled water to 1L, sterilizing at 121 ℃ for 20min, and storing for later use.
PDB medium (1L of PDB medium was prepared as an example): 200g of potato (peeled), 20g of sucrose, 1000mL of water with constant volume, sterilizing at 120 ℃ for 20min and storing for later use.
Potato integrated medium (taking 1L of potato integrated medium as an example): respectively taking 3g of monopotassium phosphate, 1.5g of magnesium sulfate heptahydrate, 1 8mg of vitamin B, 20g of glucose and 20% of potato juice until the total volume is 1000mL, sterilizing at 120 ℃ for 20min, and storing for later use; the preparation method of the potato juice comprises the following steps: removing skin from potato 200g, cutting into small pieces, adding 1000mL of water, boiling for 20min, filtering to remove potato pieces, and supplementing 1000mL of filtrate.
Wort medium (for example, 1L of wort medium was prepared): adding 4 times of water into the dry malt powder, preserving heat and saccharifying for 3-4 hours at 55-65 ℃, and checking with iodine solution until complete saccharification, thus obtaining the wort sugar. Diluting the wort sugar with water to a sugar degree of 8-10 Balin, adjusting pH to 6.0-6.5, sterilizing at 120deg.C for 20min, and storing for use.
YM medium (taking YM medium of 1L as an example): 3.0g of yeast extract, 3.0g of malt extract, 10.0g of glucose, 5.0g of peptone, 1.0L of distilled water with constant volume and pH of 6.2+/-0.2, sterilizing at 120 ℃ for 20min and storing for later use.
Soxhlet medium (taking soxhlet medium of 1L prepared as an example): 30g of sucrose, 3g of sodium nitrate, 1g of dipotassium hydrogen phosphate, 0.5g of potassium chloride, 0.5g of magnesium sulfate heptahydrate and 0.01g of ferrous sulfate are respectively taken, deionized water is added until the total volume is 1000mL, and the mixture is uniformly mixed, sterilized at 120 ℃ for 20min and then stored for standby.
EXAMPLE 1 isolation and identification of Brevibacterium
1. Separation and purification of candida bailii:
crushing a fermented grain sample of white spirit, weighing 10g of the crushed fermented grain sample, placing the crushed fermented grain sample into a triangular flask filled with 90mL of sterile distilled water, and performing shaking culture at 28 ℃ and 150rpm for 30min to obtain bacterial suspension; diluting the bacterial suspension 10 5 Taking 100 mu L of diluted bacterial suspension, uniformly coating the bacterial suspension in a Bengalia red culture medium, and culturing the bacterial suspension at 28 ℃ for 48 hours to obtain a bacterial colony with saccharomycete characteristics; placing the colony on YPD solid culture medium for streaking, separating and purifying, culturing at 28 ℃ for 48 hours, picking single colony with typical yeast colony characteristics, streaking again on YPD solid culture medium, separating and purifying again, culturing at 28 ℃ for 48 hours, and obtaining single colony which is the saccharomycete used for preparing the galactose saccharomycete fermentation sample fermentation product filtrate.
2. Identification of Brevibacterium
(1) Colony morphology and morphological observations
FIG. 1 is a colony morphology of a yeast strain isolated according to the present invention. As shown in fig. 1. The colony forms are almost consistent, the colony is fine round and white, the middle of the colony is convex, and the surface of the colony is provided with folds.
And selecting a loop of strain, and inoculating the strain into YPD liquid culture medium for culturing for 48 hours at 28 ℃ to obtain activated bacterial liquid. The activated bacterial liquid is streaked and separated on YPD solid culture medium, cultured for 48 hours at 28 ℃, a small amount of bacterial cells are taken for dyeing, the strain morphology is observed under a 1000-fold microscope, and an optical microscopic image of the strain is shown in figure 2.
(2) Molecular identification of Yeast strains
2.1 Yeast genomic DNA extraction
And (3) picking the purified saccharomycetes by using an inoculating loop, dissolving the saccharomycetes in sterile water, and uniformly mixing. Centrifuging at 6000rpm for 5min, and collecting thalli; washing thalli with phosphate buffer solution, extracting genome DNA of the washed yeast strain by using a yeast genome DNA extraction kit, measuring the concentration and purity of the extracted yeast genome DNA by a nucleic acid protein tester ND-1600, and preserving and substituting the yeast genome DNA with qualified quality under the condition of-2 ℃.
2.2 PCR amplification of target Gene
The 26rsDNA target genes were amplified using the extracted different yeast genomic DNAs as templates and NL1 and NL4 as primers, and the PCR amplification system (25 uL) included: 10xPCRbuffer2.5 uL; dNTP2.0 uL; tapDNA polymerase 0.3uL; 1.0uL of each primer; 1.0uL of DNA template; ddH2O 18.0uL. PCR reaction conditions: pre-denaturation at 95 ℃ for 5min; denaturation at 95℃for 45s; annealing at 60 ℃ for 50s; extending at 75deg.C for 1min;35 cycles; repair extension 75 ℃,1minPW. The amplified products were subjected to 1% agarose gel electrophoresis and then submitted to sequencing by Beijing qing department of biological Co., ltd, and the results after sequencing were subjected to homology comparison analysis in NCBI database.
According to the comprehensive analysis of experimental data of bacterial colony morphology and gene sequence, the isolated and purified yeast strain is identified as the genus Mortierella and named as Mortierella bainieri DGJW-58 (Trichosporon behrendii DGJW-58); the preservation number of the isolated Mortierella bainieri yeast in the China center for type culture collection is CCTCC NO: M2023647, the preservation date is 2023, 4 and 27 days, and the preservation unit address is Wuhan in China.
Example 2, galactose Yeast-like fermentation product filtrate with moisturizing, tightening and soothing effects and preparation thereof
Preparation method
The embodiment provides a preparation method of galactose yeast-like fermentation product filtrate with moisturizing, tightening and relieving effects, which comprises the following steps:
(1) Preparing fermented grains: uniformly mixing 5% of sorghum (with shell), 12% of corn, 15% of rice (with shell), 3% of glutinous rice (with shell), 5% of wheat (with shell) and 60% of water by mass to obtain a mixture. Steaming the mixture at 100deg.C for 40min, spreading, and cooling. Adding distiller's yeast 0.5% by mass, mixing, and performing solid saccharification and fermentation at 35deg.C and relative humidity of 70% for 36 hr. And filtering the fermented grains with a 200-mesh screen after the fermentation is finished, wherein the obtained filtrate is the fermented grains.
(2) Preparing seed liquid: the bacterial colony (preservation number CCTCC NO: M2023647) of the above-mentioned screened candida baileyi is inoculated into potato comprehensive culture medium by means of inoculating loop, and cultured for 48h at 28 ℃ under the condition of 150rpm of stirring paddle so as to obtain candida baileyi seed liquid.
(3) Fermentation treatment: uniformly mixing 15% of grain fermented grains, 1.5% of lactose, 5% of milk powder and 78.5% of deionized water, and sterilizing at 121 ℃ for 20min to obtain a fermentation substrate. Inoculating Mortierella bainieri seed liquor at 1% of the inoculation amount into fermentation matrix, and introducing 50dm 3 And (h L) fermenting for 24 hours at 28 ℃ by setting the rotating speed of a stirring paddle to 180 rpm.
(4) Post-treatment: the fermentation broth was placed in a high pressure homogenizer and homogenized at high pressure under 10MPa for 30min. Filtering the homogenized fermentation liquor by a diatomite filter, and sterilizing at 500MPa for 10min under high pressure to obtain galactose yeast-like fermentation product filtrate with moisturizing, tightening and relieving effects.
Example 3, galactose Yeast-like fermentation product filtrate with moisturizing, tightening and soothing effects and preparation thereof
Preparation method
Similar to example 2, except that the mass percentages of sorghum (shelled), corn, rice (shelled), glutinous rice (shelled) and wheat (shelled) in step (1) were different, the mass percentages of sorghum (shelled), 9% corn, 18% rice (shelled), 2% glutinous rice (shelled), 3% wheat (shelled) and 60% water were uniformly mixed to obtain a mixture. The remaining steps and the like have the same parameters as in example 2.
Example 4 galactose ferment with moisturizing, conditioning and soothing efficacyMother sample fungus fermentation product filtrate and its preparation
Preparation method
Similar to example 2, except that the mass percentages of sorghum (shelled), corn, rice (shelled), glutinous rice (shelled) and wheat (shelled) in step (1) were different, the mass percentages of sorghum (shelled), 13% corn, 12% rice (shelled), 5% glutinous rice (shelled), 8% wheat (shelled) and 59% water were uniformly mixed to obtain a mixture. The remaining steps and the like have the same parameters as in example 2.
Example 5, galactose Yeast-like fermentation product filtrate with moisturizing, tightening and soothing effects and preparation thereof
Preparation method
Similar to example 2, except that the mass percentages of the fermented grains, lactose milk powder and deionized water in the step (3) were different, the fermented grains with the mass percentage of 12%, lactose with the mass percentage of 2%, milk powder with the mass percentage of 8% and deionized water were uniformly mixed, and sterilized at 121 ℃ for 20min, to obtain a fermentation substrate. The remaining steps and the like have the same parameters as in example 2.
Example 6, galactose Yeast-like fermentation product filtrate with moisturizing, conditioning and soothing effects and preparation thereof
Preparation method
Similar to example 2, except that the mass percentages of the fermented grains, lactose milk powder and deionized water in the step (3) were different, the fermented grains with the mass percentages of 18%, lactose with the mass percentages of 1%, milk powder with the mass percentages of 3% and deionized water were uniformly mixed, and sterilized at 121 ℃ for 20min to obtain a fermentation substrate. The remaining steps and the like have the same parameters as in example 2.
Example 7, galactose Yeast-like fermentation product filtrate with moisturizing, tightening and soothing effects and preparation thereof
Preparation method
(1) Preparing fermented grains: mixing sorghum (with shell), corn (10%), rice (with shell) 14%, glutinous rice (with shell) 4%, wheat (with shell) 6% and water 60% to obtain mixture. Steaming the mixture at 90deg.C for 60min, spreading out, and cooling. Adding distiller's yeast 1% by mass, mixing, and performing solid saccharification and fermentation at 32deg.C and relative humidity of 89% for 24 hr. Filtering with gauze after fermentation, and collecting filtrate as grain.
(2) Preparing seed liquid: inoculating the colony of the Mortierella bainieri (preservation number CCTCC NO: M2023647) into a potato comprehensive culture medium by using an inoculating loop, and culturing for 52 hours at 30 ℃ and the rotation speed of a stirring paddle of 120rpm to obtain the Mortierella bainieri seed liquid.
(3) Fermentation treatment: uniformly mixing 16% of grain fermented grains, 1.8% of lactose, 6% of milk powder and 76.2% of deionized water, and sterilizing at 121 ℃ for 20min to obtain a fermentation substrate. Inoculating Mortierella bainieri seed liquor at 1% of the inoculation amount into fermentation matrix, and introducing 30dm 3 And (h L) fermenting for 18 hours at 30 ℃ by setting the rotating speed of a stirring paddle to 150 rpm.
(4) Post-treatment: the fermentation broth was placed in a high pressure homogenizer and homogenized at high pressure under 12MPa for 20min. Filtering the homogenized fermentation liquor with 0.45 μm, and sterilizing under 300MPa for 20min to obtain galactose yeast-like fermentation product filtrate with moisturizing, tightening and relieving effects.
Comparative example 1
Compared with example 2, comparative example 1 does not contain the process of preparing fermented grains in step (1), the fermented grains in the fermentation substrate in step (3) are replaced by lactose and milk powder in corresponding proportions (the fermentation substrate in comparative example 1 contains 4.96% of lactose, 16.54% of milk powder and 78.5% of water by mass), and the other steps have the same parameters as in example 2, so that galactose yeast-like fermentation product filtrate is prepared.
Comparative example 2
In comparison with example 2, the parameters of sorghum (with hulls), rice (with hulls), glutinous rice (with hulls), wheat (with hulls) in the mixture of step (1) of comparative example 2 are replaced by shelled sorghum, shelled rice, shelled glutinous rice, shelled wheat, the rest of the steps are the same as those of example 2, and galactose yeast-like fermentation product filtrate is prepared.
Comparative example 3
In comparison with example 2, parameters of 5% sorghum (with shell), 12% corn, 15% rice (with shell), 3% glutinous rice (with shell), 5% wheat (with shell), 60% water were changed to 12% sorghum (with shell), 5% corn, 15% rice (with shell), 3% glutinous rice (with shell), 5% wheat (with shell), 60% water, and the other steps were the same as those of example 2, to prepare a galactose yeast-like fermentation product filtrate.
Comparative example 4
In comparison with example 2, lactose in step (3) of comparative example 4 was replaced with glucose, and the remaining steps were the same as in example 2, to prepare a galactose yeast-like fermentation product filtrate.
Comparative example 5
In comparison with example 2, the preparation of a galactose yeast-like fermentation product filtrate was performed in the same manner as in example 2 except that the preparation of the galactose yeast-like fermentation product filtrate was performed in comparative example 5 by substituting the yeast of the present invention with the yeast of the present invention (from Biotechnology Co., ltd., bai. Poplar. Beijing).
Comparative example 6
Compared with example 2, the preparation method of the invention in comparative example 6 is characterized in that the preparation method is used for replacing the Mortierella bainieri with the Acremonium covering film yeast with the preservation number CCTCC M20221857 (preserved in China center for type culture collection), and the other parameters, such as the parameters of the other steps, are the same as those in example 2, so that a galactose yeast-like fermentation product filtrate is prepared.
Comparative example 7
The galactose yeast-like fermentation product filtrate was prepared using the method of application number CN202211047185.2 example 1.
Comparative example 8
In comparison with example 2, step (4) of comparative example 8 was autoclaved at 500MPa for 10min, and then incubated at 90℃for 30min, and the remaining steps were the same as in example 2, to obtain a galactose yeast-like fermentation product filtrate.
Test example one, moisture efficacy test of galactose Yeast-like fermentation product filtrate
The moisturizing effect of the galactose yeast-like fermentation product filtrate prepared in examples 2 to 7 and comparative examples 1 to 8 in the present invention was tested according to QB/T4256-2011 cosmetic moisturizing efficacy evaluation guidelines.
Test sample: galactose yeast-like fermentation product filtrate (prepared into 5% by mass aqueous solution) prepared by the methods of examples 2-7 and comparative examples 1-8, SK-II skin care essence (namely, immortal water, commercially available and undiluted), and deionized water (used as a blank control group).
Test instrument: skin stratum corneum moisture content test probe Corneometer.
Subject selection: 300 healthy volunteers aged 18-45 years (one sample was used per 20 volunteers).
The testing steps are as follows: after the face of the subject is cleaned, 1-2 mL of sample is randomly smeared on the half face area, the other half face area is used as a blank control group, and deionized water is smeared. It is used for 2 times daily, once in the morning and evening. The skin horny layer moisture content of the face skin was tested using a skin horny layer moisture content test probe before using the sample, after using the test sample for 2 weeks and 4 weeks, and the test results were averaged to calculate the rate of change of the skin horny layer moisture content. The rate of change of the moisture content of the skin stratum corneum is calculated as follows:
rate of change in skin horny layer moisture content (%) = (skin horny layer moisture content after use-skin horny layer moisture content before use)/skin horny layer moisture content before use×100%
As shown in the table above, the moisture retention of the galactose yeast-like fermentation product filtrate (5%) of the present invention was superior to that of the comparative example, and superior to SK-II skin care serum. Examples 2 to 7 have better moisture retention than comparative example 1, which shows that the fermentation process of the self-separated candida baileyi is greatly influenced by the grain, and the moisture retention effect of the prepared galactose yeast-like fermentation product filtrate is poor by changing the grain into lactose and milk powder in corresponding proportions.
The moisture retention performance of examples 2 to 7 is superior to that of comparative example 2, which shows that the preparation raw materials of the fermented grains have influence on the subsequent fermentation of the autonomously separated candida baileyi, and the fermented grains prepared from the sorghum with the shells, the rice, the glutinous rice and the wheat are more beneficial to the fermentation of yeasts, and the prepared galactose yeast-like fermentation product filtrate is rich in various nutrients, so that the moisture retention effect of the prepared galactose yeast-like fermentation product filtrate is better.
The moisture retention performance of examples 2 to 7 is superior to that of comparative example 3, which shows that the quality ratio of the preparation raw materials of the grain is more beneficial to the fermentation of saccharomycetes, the prepared galactose saccharomycetes fermentation product filtrate is rich in various nutrients, and the moisture retention effect of the prepared galactose saccharomycetes fermentation product filtrate is better.
Examples 2 to 7 have better moisture retention performance than comparative example 4, which shows that the fermentation substrate has a synergistic effect among grains, lactose and milk powder, and the preparation method of the invention has better moisture retention effect by fermenting the prepared galactose yeast-like fermentation product filtrate by using the grains, lactose and milk powder.
Examples 2 to 7 have better moisture retention performance than comparative examples 5 and 6, which shows that fermentation strains have important influence on the fermentation process, and fermentation is performed in a fermentation matrix containing grain fermented grains, lactose and milk powder by using the self-separated candida bailii of the invention, so that the prepared galactose yeast-like fermentation product filtrate has better moisture retention effect, and the obtained galactose yeast-like fermentation product filtrate has poorer moisture retention effect after being changed into other strains for fermentation.
The moisture retention properties of examples 2-7 are superior to comparative example 8, showing that the mode of sterilizing the galactose yeast-like fermentation product filtrate affects the efficacy of the final product, and the invention adopts the high-pressure sterilization mode, thereby avoiding the need of sterilizing at 80-105 ℃ in the traditional pasteurization method, and the loss of active substances in the galactose yeast-like fermentation product filtrate is easily caused by the high-temperature condition, resulting in poor moisture retention effect of the galactose yeast-like fermentation product filtrate.
The moisture retention performance of example 2 is superior to that of examples 3 to 6, which shows that the addition amount of sorghum (with shell), corn, rice (with shell), glutinous rice (with shell) and wheat (with shell) in the fermented grains, the addition ratio of the fermented grains, lactose and milk powder in the fermentation substrate has important influence on the fermentation process of saccharomycetes, and finally the moisture retention performance of the filtrate of the fermentation product of galactose saccharomycetes is influenced. In the process of preparing the fermented grains, the addition amount of sorghum (with shell), corn, rice (with shell), glutinous rice (with shell) and wheat (with shell) is preferably 3-8%, 9-13%, 12-18%, 2-5% and 3-8%. In the fermentation substrate, the addition amount of the grain fermented grains, lactose and milk powder is preferably 12-18%, 1-2% and 3-8%.
Test example II, compaction test of galactose Yeast-like fermentation product filtrate
Pore tightening effect of the galactose yeast-like fermentation product filtrate prepared in examples 2 to 7 and comparative examples 1 to 8 in the present invention was tested.
Test sample: the same as in test example 1.
Test instrument: face image analyzer VISIA-CR (Canfield, U.S.A.)
Subject selection: the same as in test example 1.
The testing steps are as follows: the testing steps are as follows: after the face of the subject is cleaned, 1-2 mL of sample is randomly smeared on the half face area, the other half face area is used as a blank control group, and deionized water is smeared. It is used for 2 times daily, once in the morning and evening. Before using the sample, after using the test sample for 2 weeks and 4 weeks, collecting the front face picture of the face of the subject, and carrying out image analysis on the cross polarized light picture to obtain the quantitative index pore area ratio, and evaluating the shrinkage condition of the face pores.
The smaller the pore area ratio in the skin, the smaller the pores in the skin, the better the pore tightening effect of the test sample.
Skin pore area ratio change rate (%) = (skin pore area ratio after use-skin pore area ratio before use)/skin pore area ratio before use×100%
As shown in the table above, the compactibility of the galactose yeast-like fermentation product filtrate (5%) of the present invention was superior to that of comparative examples 1 to 8, and the effects of the galactose yeast-like fermentation product filtrate (5%) of examples 2 to 7 were equivalent to that of SK-II skin care essence lotion for pore tightening. The effect of pore tightening of the galactose yeast-like fermentation product filtrate prepared in examples 2 to 7 is superior to that of comparative example 1, which shows that the fermentation process of the candida berosa is significantly affected by the fermented grains, and the effect of pore tightening of the galactose yeast-like fermentation product filtrate prepared by converting the fermented grains into lactose and milk powder in corresponding proportions is poor.
The pore tightening effect of the galactose yeast-like fermentation product filtrate of examples 2-7 is better than that of comparative example 2, which shows that the preparation raw materials of the grain fermented grains have influence on the subsequent fermentation of the bailia yeast, and the grain fermented grains prepared from the sorghum, rice, glutinous rice and wheat with shells are more favorable for the fermentation of the self-separated bailia yeast, and the prepared galactose yeast-like fermentation product filtrate is rich in various nutrients and has better pore tightening effect.
The pore tightening effect of examples 2-7 is superior to that of comparative example 3, which shows that the quality of the raw materials for preparing the fermented grains is more beneficial to the fermentation of the self-separated bailey yeasts, the prepared fermented grains are rich in various nutrients, and the pore tightening effect of the prepared fermented product filtrate of the galactose yeasts is better.
The pore-tightening effect of the galactose yeast-like fermentation product filtrate of examples 2 to 7 is better than that of comparative example 4, which shows that the synergy exists among the grain fermented grains, lactose and milk powder in the fermentation matrix, and the autonomously separated bailey spore yeast is fermented by the grain fermented grains, lactose and milk powder, so that the pore-tightening effect of the prepared galactose yeast-like fermentation product filtrate is better.
The pore tightening effect of the galactose yeast-like fermentation product filtrate of examples 2 to 7 is better than that of comparative examples 5 and 6, which shows that the fermentation strain has an important influence on the fermentation process, and the fermentation process is performed by using the independently separated bailia baileyi yeast of the invention in a fermentation matrix containing grain fermented grains, lactose and milk powder, so that the pore tightening effect of the prepared galactose yeast-like fermentation product filtrate is better, and the obtained galactose yeast-like fermentation product filtrate is changed into other strains for fermentation, so that the pore tightening effect of the obtained galactose yeast-like fermentation product filtrate is poorer.
The pore tightening effect of the galactose yeast-like fermentation product filtrate of examples 2-7 is superior to that of comparative example 8, which shows that the sterilization mode of the galactose yeast-like fermentation product filtrate affects the efficacy of the final product.
The pore tightening effect of the galactose yeast-like fermentation product filtrate prepared in example 2 is superior to that of examples 3 to 6, which shows that the addition amount of sorghum (with shell), corn, rice (with shell), glutinous rice (with shell) and wheat (with shell) in the fermented grains, the addition ratio of the fermented grains, lactose and milk powder in the fermentation matrix has an important influence on the fermentation process of the saccharomyces bailii, and finally the pore tightening effect of the galactose yeast-like fermentation product filtrate is influenced. In the process of preparing the fermented grains, the addition amount of sorghum (with shell), corn, rice (with shell), glutinous rice (with shell) and wheat (with shell) is preferably 3-8%, 9-13%, 12-18%, 2-5% and 3-8%. In the fermentation substrate, the addition amount of the grain fermented grains, lactose and milk powder is preferably 12-18%, 1-2% and 3-8%.
Test example III relief efficacy test of galactose Yeast-like fermentation product filtrate
The test principle is as follows: neutrophils of zebra fish embryos and human neutrophils are highly similar in morphology, biochemistry and physiological functions, the neutrophils are white blood cells appearing in the first place at a damaged or germ invasion part, the neutrophils act on a model for eliminating infection or harmful substances, the model is used for testing, the number change of the neutrophils in the fish embryo lateral line area of a test object treatment group and a model control group is compared, and the neutrophil inhibition rate is calculated to evaluate the relieving efficacy of a test sample, wherein the model is used for inducing the nerve hillock cell damage in the zebra fish embryo lateral line area to cause the neutrophil aggregation.
Blank control group: 24 healthy zebra fish embryos 3 days after fertilization were randomly selected, transferred to a 3cm dish, and 5mL of copper sulfate pentahydrate (CuSO) containing 0.16mg/L was added 4 ·5H 2 The fish embryo culture solution of O); positive control group: 24 healthy zebra fish embryos 3 days after fertilization were randomly selected, transferred to a 3cm dish, and 5mL of copper sulfate pentahydrate (CuSO) containing 0.16mg/L was added 4 ·5H 2 O) and 0.0036mg/L indomethacin fish embryo culture solution (indomethacin has a relieving effect, can inhibit neutrophil aggregation, and a positive control is used for judging the effectiveness of the experiment. ).
Experimental group: dissolving the galactose yeast fermentation product filtrate prepared in examples 2-7 and comparative examples 1-8 in fish embryo culture solution to prepare a sample solution with the mass percent of 3%, randomly selecting 24 healthy zebra fish embryos 3 days after fertilization, and transferringTo a 3cm dish, 5mL of copper sulfate pentahydrate (CuSO) containing 0.16mg/L was added 4 ·5H 2 O) and a fish embryo culture solution of a sample to be tested.
The testing method comprises the following steps: placing the blank control group, the positive control group and the experimental group in a constant temperature oven at 28deg.C for culturing for 40min, fixing each tested group of fish embryo in paraformaldehyde solution (4%) for at least 1 hr, treating fish embryo with PBST (PBS solution with mass percentage of Triton X-100 of 0.04%) for 3 times and 5min each time, treating with 50% ethanol for 3min, staining fish embryo with Sudan black (0.4%) staining solution at room temperature for 1 hr, soaking with 70% ethanol for 4 times and 5min each time, treating with PBST for 2 times and 5min each time, and treating with bleaching solution (H) 2 O 2 And KOH for 3% and 1% by mass respectively, then 5min with 70% ethanol solution, 1min with PBST, 15min with clear solution 1 (20% and 0.25% by mass respectively) and 10min with clear solution 2 (50% and 0.25% by mass respectively) and 3min with PBST, placing fish embryos sideways, placing them in a split microscope, counting the number of neutrophils in the three-quarter tail side region of each fish embryo from the hepatic portal, and calculating the neutrophil aggregation inhibition rate by the following formula, the results of the inhibition of the neutrophil aggregation of zebra fish embryos by galactose yeast-like fermentation product filtrate (3 wt%) of examples 2 to 7 and comparative examples 1 to 8 are shown in the following table.
Wherein T represents the average value of the number of the neutrophils in the zebra fish embryo of the experimental group; c represents the average value of the number of neutrophils in zebra fish embryos of the blank group.
The effect of the galactose yeast fermentation product filtrate prepared in examples 2 to 7 was superior to that of comparative example 1, which shows that the fermented grains have an important effect on the fermentation process of the Brevibacterium, and that the effect of the prepared galactose yeast-like fermentation product filtrate was poor by changing the fermented grains into lactose and milk powder in the corresponding proportions.
The effect of relieving the galactose yeast fermentation product filtrate prepared in examples 2-7 is superior to that of comparative example 2, which shows that the preparation raw materials of the fermented grains have influence on the subsequent fermentation of the bailia yeast, and the fermented grains prepared from the sorghum, rice, glutinous rice and wheat with shells are more beneficial to the fermentation of the bailia yeast, and the prepared galactose yeast-like fermentation product filtrate is rich in various nutrients, and the prepared galactose yeast-like fermentation product filtrate has better effect of relieving.
The soothing effect of examples 2 to 7 is superior to that of comparative example 3, which shows that the quality of the raw materials for preparing the fermented grains is more favorable for the fermentation of the bailing candida and the filtrate of the fermented product of the prepared galactose yeast is rich in various nutrients, and the soothing effect of the filtrate of the fermented product of the prepared galactose yeast is better as 3 to 8 percent of sorghum (with a shell), 9 to 13 percent of corn, 12 to 18 percent of rice (with a shell), 2 to 5 percent of glutinous rice (with a shell) and 3 to 8 percent of wheat (with a shell) are used for influencing the subsequent fermentation of the bailing candida which is independently separated.
The soothing effect of the galactose yeast fermentation product filtrate prepared in examples 2-7 is superior to that of comparative example 4, which shows that in the fermentation substrate, the grain, lactose and milk powder have a synergistic effect, the autonomously separated bailey spore yeast is fermented by using the grain, lactose and milk powder, and the soothing effect of the galactose yeast-like fermentation product filtrate prepared is better.
The relief performance of the galactose yeast fermentation product filtrate prepared in examples 2-7 is superior to that of comparative examples 5 and 6, which shows that the fermentation strain has an important influence on the fermentation process, and the galactose yeast-like fermentation product filtrate prepared by fermenting the above-mentioned fermentation substrate containing grain, lactose and milk powder with the above-mentioned independently-separated Brevibacterium flavum has a better relief effect, and is changed into other strains for fermentation, so that the relief effect of the obtained galactose yeast-like fermentation product filtrate is poor.
The soothing performance of the galactose yeast fermentation product filtrate prepared in examples 2-7 is superior to that of comparative example 8, which shows that the sterilization mode of the galactose yeast fermentation product filtrate influences the efficacy of the final product.
The effect of relieving the galactose yeast fermentation product filtrate prepared in example 2 is superior to that of examples 3 to 6, which shows the addition amount of sorghum (with shell), corn, rice (with shell), glutinous rice (with shell) and wheat (with shell) in the fermented grains, the addition ratio of the fermented grains, lactose and milk powder in the fermentation matrix, and has important influence on the fermentation process of saccharomycetes, and finally the effect of relieving the galactose yeast-like fermentation product filtrate. In the process of preparing the fermented grains, the addition amount of sorghum (with shell), corn, rice (with shell), glutinous rice (with shell) and wheat (with shell) is preferably 3-8%, 9-13%, 12-18%, 2-5% and 3-8%. In the fermentation substrate, the addition amount of the grain fermented grains, lactose and milk powder is preferably 12-18%, 1-2% and 3-8%.
Test example IV, repair efficacy test of galactose Yeast-like fermentation product filtrate
The repair effect of the galactose yeast-like fermentation product filtrate prepared in examples 2 to 7 and comparative examples 1 to 8 in the present invention was tested.
Test sample: the same as in test example 1.
Test instrument: skin moisture loss test probe (Tewameter TM 330T); skin pigment test probe (Mexameter MX 18).
Subject selection: the same as in test example 1.
The testing steps are as follows: the testing steps are as follows: after the face of the subject is cleaned, 1-2 mL of sample is randomly smeared on the half face area, the other half face area is used as a blank control group, and deionized water is smeared. It is used for 2 times daily, once in the morning and evening. The skin moisture loss test probe was used to test the skin moisture loss of the face and the red pigment content of the reddish skin area before and after 2 and 4 weeks of sample application.
Proper skin moisture evaporation is normal skin formation metabolism, the greater the skin's transdermal moisture loss, the more severe the skin moisture evaporation per unit time, the poorer the skin's barrier function. When the skin moisture loss becomes small after the sample is used, it is indicated that the skin water evaporation amount per unit time becomes small and the skin barrier function is restored.
The higher the skin haematochrome content, the more serious the skin redness, and if the sample is used, the lower the skin haematochrome content is, the improvement of the skin redness and the repair of the skin barrier function are shown.
Skin transdermal moisture change rate (%) = (skin transdermal moisture loss after use-skin transdermal moisture loss before use)/skin transdermal moisture loss before use×100%;
skin red pigment content change rate (%) = (skin red pigment content after use-skin red pigment content before use)/skin red pigment content before use×100%.
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As shown in the table above, after the galactose yeast-like fermentation product filtrate (5%) of the present invention is used, the percutaneous moisture loss of the facial skin of the subject is reduced, and the red pigment content of the skin reddish region is reduced, which indicates that the galactose yeast-like fermentation product filtrate of the present invention has excellent barrier repairing and reddish improving effects, i.e., the galactose yeast-like fermentation product filtrate of the present invention has excellent repairing effects. Among them, the repairing effect of example (5%) is better than that of comparative example (5%), and is better than SK-II skin care essence.
The repair effect of the galactose yeast-like fermentation product filtrate prepared in examples 2 to 7 is superior to that of comparative example 1, which shows that the fermented grains have important influence on the fermentation process of yeasts, and the repair effect of the galactose yeast-like fermentation product filtrate prepared by changing the fermented grains into lactose and milk powder in corresponding proportions is poor.
The repairing effect of the galactose yeast-like fermentation product filtrate prepared in examples 2-7 is better than that of comparative example 2, which shows that the preparation raw materials of the fermented grains have influence on the subsequent fermentation of the candida baileyi, and the fermented grains prepared from the sorghum, rice, glutinous rice and wheat with shells are more beneficial to the fermentation of the yeasts, and the prepared galactose yeast-like fermentation product filtrate is rich in various nutrients and has better repairing effect.
The repairing effect of examples 2-7 is better than that of comparative example 3, which shows that the quality of the raw materials for preparing the fermented grains is more beneficial to the fermentation of saccharomycetes, the filtrate of the fermented product of the prepared galactose saccharomycetes is rich in various nutrients, and the repairing effect of the filtrate of the fermented product of the prepared galactose saccharomycetes is better.
The repair effect of the galactose yeast-like fermentation product filtrate prepared in examples 2-7 is better than that of comparative example 4, which shows that the synergy exists among the grain, lactose and milk powder in the fermentation matrix, the autonomously separated bailey spore yeast is fermented by the grain, lactose and milk powder, and the repair effect of the prepared galactose yeast-like fermentation product filtrate is better.
The repair effect of the galactose yeast-like fermentation product filtrate prepared in examples 2 to 7 is better than that of the galactose yeast-like fermentation product filtrate prepared in comparative examples 5 and 6, which shows that the fermentation strain has an important influence on the fermentation process, the galactose yeast-like fermentation product filtrate prepared by fermenting the above-mentioned fermentation substrate containing grain fermented grains, lactose and milk powder with the above-mentioned fermentation substrate has a better repair effect, and the repair effect of the galactose yeast-like fermentation product filtrate obtained by fermenting the above-mentioned fermentation substrate with other strains is worse.
The repair effect of the galactose yeast-like fermentation product filtrate prepared in examples 2-7 is superior to that of comparative example 8, which shows that the sterilization mode of the galactose yeast-like fermentation product filtrate affects the efficacy of the final product.
The repairing effect of the galactose yeast-like fermentation product filtrate prepared in example 2 is superior to that of examples 3 to 6, which shows the addition amount of sorghum (with shell), corn, rice (with shell), glutinous rice (with shell) and wheat (with shell) in the fermented grains, the addition ratio of the fermented grains, lactose and milk powder in the fermentation matrix has an important influence on the fermentation process of the candida berosa, and finally the repairing effect of the galactose yeast-like fermentation product filtrate is influenced. In the process of preparing the fermented grains, the addition amount of sorghum (with shell), corn, rice (with shell), glutinous rice (with shell) and wheat (with shell) is preferably 3-8%, 9-13%, 12-18%, 2-5% and 3-8%. In the fermentation substrate, the addition amount of the grain fermented grains, lactose and milk powder is preferably 12-18%, 1-2% and 3-8%.
Test example five skin irritation test of galactose Yeast-like fermentation product filtrate
The specific method for testing the skin irritation of the invention is carried out by referring to the standard in cosmetic safety technical Specification (2015 edition), and the specific method is as follows:
24 white rabbits satisfying the test conditions were prepared (4 white rabbits were used for each sample), and the hair on both sides of the back vertebra of the rabbits was cut off (the hair removal ranges were 3 cm. Times.3 cm each) before the test.
0.5mL of the test substance (galactose yeast-like fermentation product filtrate of examples 2 to 7) was applied to one side of the skin (application area was 2.5 cm. Times.2.5 cm), and the other side was not treated as a control. The application was once daily for 14 days. From the next day, the hair was sheared before each application. The residual test substance was removed with pure water. After 1h the results were observed.
The control and test areas were treated as described in the cosmetic safety Specification (2015 edition) with skin prick/corrosiveness test Table 1.
Evaluation of results: the average integral of each rabbit of the rabbits per day was calculated according to the following formula, and the skin irritation intensity of the rabbits was judged according to the skin irritation/corrosiveness test sheet of cosmetic safety Specification (2015). In the test, it was observed whether or not the skin had symptoms other than skin irritation.
Average per animal per day = (Σintegral of erythema and edema/number of animals tested)/14
The test results are shown in the following table.
| Sample of | Mean of the integral of each animal per day | Results |
| Control group | 0 | No skin irritation |
| Example 2 | 0 | No skin prickExcitation property |
| Example 3 | 0 | No skin irritation |
| Example 4 | 0 | No skin irritation |
| Example 5 | 0 | No skin irritation |
| Example 6 | 0 | No skin irritation |
| Example 7 | 0 | No skin irritation |
Test results show that the galactose yeast-like fermentation product filtrate prepared in the embodiments 2-7 has good moisturizing, tightening and relieving effects, does not have skin irritation, and is safe to use.
Finally, it should be noted that the above embodiments are only for illustrating the technical solution of the present invention and not for limiting the scope of the present invention, and although the present invention has been described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that the technical solution of the present invention may be modified or substituted equally without departing from the spirit and scope of the technical solution of the present invention.
Claims (11)
1. A method for preparing galactose yeast-like fermentation product filtrate with moisturizing, tightening and relieving effects, which is characterized by comprising the following steps:
S1, preparing fermented grains: uniformly mixing sorghum, corn, rice, glutinous rice, wheat and water to obtain a mixture, steaming, cooling, adding distiller's yeast for saccharification and fermentation, and filtering to obtain fermented grains;
s2, preparing seed liquid: inoculating bacterial colonies of the candida berosa to a liquid culture medium for culture to obtain seed liquid;
s3, fermentation treatment: uniformly mixing the fermented grains obtained in the step S1, lactose, milk powder and water, and sterilizing to obtain a fermentation substrate; inoculating the seed liquid in the step S2 into the fermentation substrate for fermentation to obtain fermentation liquid;
s4, post-processing: carrying out high-pressure homogenization treatment, filtration treatment and sterilization treatment on the fermentation broth in the step S3 to obtain galactose yeast-like fermentation product filtrate with moisturizing, tightening and relieving effects;
the Brevibacterium is preserved in China Center for Type Culture Collection (CCTCC) in the period of 2023, 4 and 27, and the preservation number is M2023647;
in the step S1, the mixture comprises 3-8% of sorghum, 9-13% of corn, 12-18% of rice, 2-5% of glutinous rice, 3-8% of wheat and the balance of water in percentage by mass;
in the step S3, the fermentation substrate comprises 12-18% of grain fermented grains, 1-2% of lactose, 3-8% of milk powder and the balance of water in percentage by mass;
The sterilization treatment is to subject the filtered fermentation liquor to high-pressure sterilization for 5-20 min under 300-800 MPa.
2. The method for preparing a galactose yeast-like fermentation product filtrate as set forth in claim 1, wherein in step S1, the mixture comprises 5% sorghum, 12% corn, 15% rice, 3% glutinous rice, 5% wheat, and the balance water in mass percent.
3. The method for preparing a galactose yeast-like fermentation product filtrate according to claim 1, wherein sorghum, rice, glutinous rice and wheat are all shelled.
4. The method for producing a galactose yeast-like fermentation product filtrate as set forth in claim 1, wherein in step S1, the distiller' S yeast is added in an amount of 0.2 to 1% by mass of the mixture.
5. The method for producing a galactose yeast-like fermentation product filtrate as set forth in claim 4, wherein the distiller's yeast is added in an amount of 0.5% by mass of the mixture.
6. The method for preparing a galactose yeast-like fermentation product filtrate as set forth in claim 1, wherein in the step S1, the temperature of the saccharification and fermentation is 32-38 ℃, the time of the saccharification and fermentation is 24-38 hours, and the relative humidity of the saccharification and fermentation is 60-80%.
7. The method for preparing a galactose yeast-like fermentation product filtrate as set forth in claim 1, wherein in step S3, the fermentation substrate comprises 15% of grain, 1.5% of lactose, 5% of milk powder, and the balance of water in mass percent.
8. The method for preparing a galactose yeast-like fermentation product filtrate according to claim 1, wherein in the step S3, the fermentation temperature is 25 to 32 ℃ and the fermentation time is 18 to 36 hours.
9. The method for producing a galactose yeast-like fermentation product filtrate according to claim 1, wherein in the step S4, the high-pressure homogenization treatment is to homogenize the fermentation broth under a high pressure of 8 to 12MPa for 20 to 60 minutes.
10. The galactose yeast-like fermentation product filtrate with moisturizing, tightening and soothing effects prepared by the method for preparing the galactose yeast-like fermentation product filtrate as claimed in any one of claims 1 to 9.
11. Use of the galactose yeast like fermentation product filtrate with moisturizing, tightening and soothing effects as claimed in claim 10 for preparing cosmetics.
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| CN102382810A (en) * | 2011-10-28 | 2012-03-21 | 石家庄金太阳生物有机肥有限公司 | Preparation method of probiotic inoculum for preventing replant disease of greenhouse cucumbers |
| CN115125153A (en) * | 2022-08-30 | 2022-09-30 | 广州优科生物科技有限公司 | Preparation method and application of galactose yeast-like bacteria fermentation product filtrate |
| CN115161207A (en) * | 2022-08-24 | 2022-10-11 | 宁波格鲁康生物科技有限公司 | Preparation method of galactose yeast solution, galactose yeast powder and application of galactose yeast powder in skin brightening and wrinkle diminishing |
| CN115607483A (en) * | 2022-09-15 | 2023-01-17 | 广州市小谭科技有限公司 | Yeast fermentation composition with multiple effects and application thereof |
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| CN102382810A (en) * | 2011-10-28 | 2012-03-21 | 石家庄金太阳生物有机肥有限公司 | Preparation method of probiotic inoculum for preventing replant disease of greenhouse cucumbers |
| CN115161207A (en) * | 2022-08-24 | 2022-10-11 | 宁波格鲁康生物科技有限公司 | Preparation method of galactose yeast solution, galactose yeast powder and application of galactose yeast powder in skin brightening and wrinkle diminishing |
| CN115125153A (en) * | 2022-08-30 | 2022-09-30 | 广州优科生物科技有限公司 | Preparation method and application of galactose yeast-like bacteria fermentation product filtrate |
| CN115607483A (en) * | 2022-09-15 | 2023-01-17 | 广州市小谭科技有限公司 | Yeast fermentation composition with multiple effects and application thereof |
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