CN110772459B - Yeast fermented birch juice and application thereof in whitening cosmetic composition - Google Patents
Yeast fermented birch juice and application thereof in whitening cosmetic composition Download PDFInfo
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- 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
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- A61K8/9789—Magnoliopsida [dicotyledons]
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- 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
- A61Q19/02—Preparations for care of the skin for chemically bleaching or whitening the skin
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- 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
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Abstract
The present invention relates to a yeast fermented birch sap obtained by fermentation using yeast as a strain in a medium comprising birch sap and optionally fruit powder. The present invention also relates to a method for producing yeast fermented birch juice comprising the step of fermenting in a medium comprising birch juice and optionally fruit powder using yeast as a strain.
Description
Technical Field
The present invention relates to a yeast fermented birch juice and its use in whitening cosmetic compositions, wherein the yeast fermented birch juice is obtained by fermentation using yeast as a strain in a medium comprising birch juice and optionally fruit powder.
Background
Birch sap (also called birch sap) is fresh sap obtained by cutting bark or drilling trunk of birch, contains saccharide, amino acid, vitamins, biotin, trace mineral elements, aromatic oil, betulin, saponin and other compounds, and has good skin care effects of moisturizing, resisting inflammation, removing wrinkle, whitening and the like. The yeast lysate generated after the fermentation of the birch juice by the yeast is rich in amino acid, polypeptide and various vitamins, and has certain whitening and moisturizing effects.
The application of the birch juice fermented by the yeast is limited in the field of wines and beverages, and the process aims to increase the content of ethanol in the birch juice, but the birch juice cannot have other active ingredients or improve the efficacy of the birch juice. For example, CN107586640A discloses a fermentation process of birch sap wine, CN108676666A discloses a distilled birch sap wine and its production process, and CN108676666A discloses a beer of birch sap and its brewing method.
At present, the report that the birch juice fermented by the yeast is used in the field of cosmetics does not exist. CN109077960A discloses an anti-aging and whitening composition of birch juice, wherein fermented birch juice is used as cosmetic raw material, but the fermentation strain, functional components and efficacy of the raw material are unknown, and the problems of long fermentation period of 30 days, low production efficiency and the like exist.
In order to widen the application of the fermented birch juice, the inventor adopts saccharomycetes as a strain, takes the birch juice as a main substrate, optionally adds a growth promoting component such as fruit powder into the birch juice, and performs a fermentation process, so that the obtained saccharomycetes-fermented birch juice has excellent whitening effect.
Disclosure of Invention
The inventor finds that the yeast is used as a strain and the birch juice is used as a main substrate, and optionally fruit powder is added into the substrate for fermentation, so that a yeast fermented birch juice product with improved performance relative to the birch juice can be obtained, and the yeast fermented birch juice product has the active components of the birch juice and is rich in yeast cell components and active components generated by a fermentation process, such as polyphenol, so that the product has excellent whitening effect and can be used as a whitening active raw material of a whitening cosmetic composition.
Thus, in one aspect, the present invention relates to a method for producing yeast fermented birch juice comprising the step of performing fermentation in a medium comprising birch juice and optionally fruit powder using yeast as a strain.
In one embodiment, the method comprises the steps of:
(1) Fermenting in a culture medium containing birch juice and optional fruit powder by using yeast as a strain to obtain a fermentation liquid product;
(2) Centrifuging and filtering the fermentation product to respectively obtain saccharomycete and fermented birch juice filtrate;
(3) Crushing the yeast thalli under high pressure, and then carrying out centrifugal filtration to obtain soluble yeast lysate serving as supernatant and yeast thalli fragments; and
(4) Mixing the obtained yeast lysate with the obtained fermented birch juice filtrate, and filtering to obtain fermented birch juice filtrate (referred to as "yeast fermented birch juice").
The yeast used in the invention can comprise saccharomyces cerevisiae, saccharomyces boulardii, yarrowia lipolytica and the like, wherein the saccharomyces cerevisiae is preferred. Such yeasts are commercially available in the form of dried yeasts, for example, from Dupont Danisco, angel Yeast, inc; alternatively, the yeast may be obtained by expanding a wild yeast strain.
The birch juice is obtained from Betula genus of Betulaceae family, and can be derived from Betula platyphylla (Betula alba), betula tenella (Betula pubescens), betula Pendula (Betula Pendula) and Betula platyphylla (Betula platyphylla). The birch juice is colorless and transparent, has no precipitate and impurities, and has faint scent and rich nutrition of birch, and is collected by manually drilling holes at the base of the trunk of the birch between thawing and early spring leaf emergence. The birch juice is commercially available and used as such, for example from greater Khingan over wild berry development, LLC.
Birch sap that may be used in the present invention includes birch sap stock solution or concentrated birch sap (also known as birch sap concentrate). Wherein the concentration ratio of the concentrated birch sap is usually about 1.2-10 times, preferably about 1.5-4 times.
The concentrated birch sap is obtained by concentrating a commercially available birch sap product. Concentration methods are known in the art, such as heat concentration, low temperature vacuum concentration, membrane concentration, and the like. In the present invention, it is preferred to perform concentration by low temperature freeze concentration or membrane concentration process, for example, commercially available birch juice stock solution is introduced into a low temperature drying device, cooled to about-40 ℃ to-70 ℃, and subjected to low temperature vacuum concentration by vacuumizing to about 0.1-30Pa, thereby obtaining concentrated birch juice with different concentration times.
The above step (1) fermentation is known in the art. For example, the birch sap culture medium is added to about 100-12000L fermenter in an amount of about 60-70% (v/v) and sterilized at about 105 ℃ for about 20-30 minutes; taking fermentation culture medium volume as a standard, inoculating yeast seed liquid into a fermentation tank under an aseptic condition, stirring at the temperature of about 25-30 ℃ and the rotating speed of about 180-350rmp, continuously fermenting for about 6-48 hours under the ventilation quantity of about 1.2-2.0vvm, and stopping the tank to finish the fermentation process to obtain a fermentation liquid product.
Wherein the yeast seed solution is obtained by rehydrating active dry yeast. Rehydration of active dry yeast is known in the art, for example, by weighing active dry yeast in an amount of about 0.1-0.8g/L, preferably about 0.3-0.5g/L, of birch juice, adding to about 5-15 times the volume of sterile water, and rehydrating at about 28-35 ℃ for about 15-20 minutes to obtain yeast seed solution. Alternatively, yeast seed solutions can be obtained by expanding wild yeast in culture medium, such cultures being known in the art.
The birch juice culture medium can be prepared by using birch juice, especially concentrated birch juice, such as concentrated birch juice 1.5-4 times as substrate. The birch sap content in the medium is above about 93%, preferably above about 95%, based on the total weight of the birch sap medium.
Preferably, fruit powder as a carbon source may be added to the birch sap culture medium. The fruit powder includes, but is not limited to, apple powder, litchi powder, peach powder, cherry powder, roxburgh rose powder, grape powder and mixtures thereof, wherein cherry powder is preferred. The content of the fruit powder in the birch juice medium is generally about 0-5%, preferably about 0.5-3%, based on the total weight of the birch juice medium.
Further, a pH adjusting agent may be added to the birch sap culture medium to adjust the pH of the birch sap culture medium to about 4.5-5.0. Such pH adjusting agents are known in the art and include, for example, but are not limited to, lactic acid, citric acid, sodium lactate, sodium citrate, and sodium hydroxide, with sodium citrate being preferred.
In addition, inorganic salts that promote yeast growth may also be added to the medium, examples of which include, but are not limited to, potassium dihydrogen phosphate, anhydrous magnesium sulfate, sodium dihydrogen phosphate, and the like. The inorganic salts are used in amounts known in the art, for example, about 0.05-0.2% based on the total weight of the birch juice medium.
The birch juice culture medium can be prepared by adding fruit powder, pH regulator and/or inorganic salt into birch juice.
Centrifugation and filtration of the resulting fermentation product of step (2) above are known in the art. Typically, this is done for about 15-30 minutes at about 6000-10000rpm, for example, on a high speed centrifuge. The centrifugation and filtration step separates yeast thallus from the fermented birch juice filtrate (supernatant).
The yeast cells obtained by the high-pressure disruption in the step (3) are known in the art. For example, the yeast cells obtained are resuspended using 3-5 times the mass of fermented birch juice to obtain a suspension of yeast cells, which is then uniformly treated with a high-pressure cell crusher at a flow rate of about 1-2 liters/minute and a pressure of about 1000-1200 bar until the disruption rate of the yeast cells reaches about 95% or more. Then, the disrupted yeast cell fluid is centrifuged and filtered, usually at about 6000 to 10000rpm for about 15 to 30 minutes, to obtain a soluble yeast lysate as a supernatant, and yeast cell debris.
In the step (4), the soluble yeast lysate obtained in the step (3) and the fermented birch juice filtrate obtained in the step (2) are uniformly mixed and then filtered, and the obtained filtrate is the birch juice filtrate product fermented by yeast (referred to as the yeast fermented birch juice for short).
The method may further comprise subjecting the resultant product to ultra-high temperature flash sterilization, wherein the sterilization temperature is about 115 ± 5 ℃ for about 10-30 seconds; the sterilized product is then transferred to a storage tank for storage.
The obtained yeast fermented birch juice is light and transparent, and contains nutrients of birch juice substrate (including B vitamins, trace elements, amino acids, fatty acid, etc.), nutrients of soluble yeast lysate (including polysaccharide, protein, nucleic acid, etc.), and polyphenols obtained from fermentation of birch juice. Typically, the yeast-fermented birch juice comprises about 450-1200mg/L polyphenols, about 4-8g/L polysaccharides, about 0.5-1.5% proteins, and about 600-1100mg/L nucleic acids. Therefore, the obtained yeast fermented birch juice contains abundant active nutrients, can be used as a raw material nutrient for whitening cosmetics, and has excellent whitening effect.
In another aspect, the present invention relates to a yeast fermented birch juice obtained by fermentation using yeast as a bacterial species in a medium comprising birch juice and optionally fruit powder.
Typically, the yeast-fermented birch juice comprises about 450-1200mg/L polyphenols, about 4-8g/L polysaccharides, about 0.5-1.5% proteins, and about 600-1100mg/L nucleic acids.
In yet another aspect, the present invention relates to the use of said yeast fermented birch juice in whitening cosmetic compositions.
In still another aspect, the present invention relates to a whitening cosmetic composition comprising (a) yeast fermented birch juice.
The content of the yeast-fermented birch juice in the whitening cosmetic composition may vary within wide ranges, for example, from more than 0 to less than 100%, preferably about 20-95%, based on the total weight of the whitening cosmetic composition.
The whitening cosmetic composition may optionally include (B) ingredients commonly used in whitening cosmetic compositions, including but not limited to vehicles, active ingredients, and adjuvants, etc., in addition to the yeast-fermented birch juice. Component (B) is known in the art, and the type and amount thereof can be selected by those skilled in the art as needed, for example, the content of component (B) is about 2 to 82% based on the total weight of the whitening cosmetic composition.
The vehicle includes, for example, diluents, dispersants or carriers and the like, examples of which include, but are not limited to, ethanol, dipropylene glycol, butylene glycol, and the like. The content of the vehicle in the whitening cosmetic composition is known in the art, and for example, it is generally about 0.5 to 20% by weight of the total weight of the component (B).
Such active ingredients include, for example, emollients, humectants, whitening actives and the like.
Examples of such emollients include, but are not limited to, one or more of olive oil, macadamia nut oil, sweet almond oil, grape seed oil, avocado oil, corn oil, sesame oil, soybean oil, peanut oil, meadowfoam seed oil, safflower seed oil, rosa canina oil, argan oil, jojoba oil, sunflower seed oil, oil of mauritika palm, squalane, ethylhexyl palmitate, isopropyl myristate, hydrogenated polyisobutene, isohexadecane, isododecane, diethylhexyl carbonate, dioctyl carbonate, isopropyl lauroyl sarcosinate, isononyl isononanoate, hydrogenated polydecene, tris (ethylhexanoate), cetyl ethylhexanoate, bis-diethoxydiethylene glycol cyclohexane 1, 4-dicarboxylate, caprylic/capric triglyceride, oleyl erucate, octyldodecanol myristate, octyldodecanol, dimethicone, octylmethicone, cetyldimethicone, cyclopentadimethicone, and the like. Examples of solid emollients include, but are not limited to, one or more of cetyl alcohol, stearyl alcohol, cetostearyl alcohol, behenyl alcohol, batyl alcohol, lauric acid, myristic acid, palmitic acid, stearic acid, beeswax, candelilla wax, carnauba wax, lanolin, ozokerite wax, jojoba seed wax, paraffin wax, microcrystalline wax, hydrogenated rice bran wax, hydrogenated coconut oil glycerides, glyceryl behenate/eicosanoate, myristyl myristate, bis-diglycerol polyacyladipate-2, shea butter, pluronic palm seed fat, and the like. The amount of said emollient in said whitening cosmetic composition is known in the art and is for example generally comprised between about 1 and 50% of the total weight of component (B).
Examples of such humectants include, but are not limited to, one or more of glycerol, diglycerol, butylene glycol, propylene glycol, 1, 3-propanediol, dipropylene glycol, 1, 2-pentanediol, polyethylene glycol-8, polyethylene glycol-32, methyl gluceth-10, methyl gluceth-20, PEG/PPG-17/6 copolymer, glyceryl polyether-7, glyceryl polyether-26, glyceryl glucoside, PPG-10 methyl glucose ether, PPG-20 methyl glucose ether, PEG/PPG/polytetramethylene glycol-8/5/3 glycerol, sucrose, trehalose, rhamnose, mannose, raffinose, betaine, erythritol, xylitol, urea, glyceryl polyether-5 lactate, sodium hyaluronate, hydrolyzed sodium hyaluronate, sodium hyaluronate acetylated, sodium polyglutamate, hydrolyzed sclerotium gum, pullulan, tremella polysaccharide, tamarind seed polysaccharide, and the like. The content of the humectant in the whitening cosmetic composition is known in the art, and for example, it is generally about 1 to 30% by weight based on the total weight of the component (B).
The whitening active ingredients include, but are not limited to, one or more of kojic acid, ascorbyl glucoside, arbutin, tranexamic acid, niacinamide, phytosterols/behenyl alcohol/octyldecanol lauroyl glutamate, phenethyl resorcinol, turmeric root extract, birch bark extract, ceramide 2, ceramide 3, acetyl phytosphingosine, resveratrol, palmetto bark extract, coleus forskohlii root extract, pepper seed extract, ubiquinone, cholesterol stearate, ascorbic acid, ascorbyl dipalmitate, tocopherol (vitamin E), tocopheryl acetate, bisabolol, ascorbyl tetraisopalmitate, pyridoxine dicaprylate, pyridoxine dipalmitate, retinol palmitate, phytosterols/octyldodecanol lauroyl glutamate, bis-behenyl alcohol/isostearyl alcohol/phytosterol dimer linoleate, phytosterols oleate, macadamia oleate, various plant extracts, and the like. The content of the whitening ingredient in the whitening cosmetic composition is known in the art, and for example, it is generally about 0.01 to 30% by weight based on the total weight of the component (B).
Such adjuvants include, for example, emulsifiers, thickeners, preservatives, fragrances and the like.
Examples of such emulsifiers include, but are not limited to, one or more of cetearyl olivate, sorbitan olivate, polysorbate-60, polysorbate-80, methylgluco sesquistearate, PEG-20 methylgluco sesquistearate, PEG-40 hydrogenated castor oil, PPG-26-butanoeth-26, PEG-4 polyglyceryl-2 stearate, PEG-60 hydrogenated castor oil, steareth-2, steareth-21, PPG-13-decyltetradecyl polyether-24, cetearyl glucoside, PEG-100 stearate, glyceryl stearate SE, coco glucoside, cetearyl polyether-25, PEG-40 stearate, polyglyceryl-3 methylgluco distearate, glyceryl stearate citrate, polyglyceryl-10 stearate, polyglyceryl-10 myristate, polyglyceryl-10 dioleate, polyglyceryl-10 laurate, polyglyceryl-10 isostearate, polyglyceryl-10 oleate, polyglyceryl-10 diisostearate, polyglyceryl-6 laurate, polyglyceryl-6 myristate, sucrose, polystearate, and the like. The content of the emulsifier in the whitening cosmetic composition is known in the art, and for example, it is generally about 0.5 to 10% by weight based on the total weight of the component (B).
Examples of the thickener include, but are not limited to, one or more of carbomers, acrylates and derivatives thereof, xanthan gum, acacia, polyethylene glycol-14M, polyethylene glycol-90M, succinoglycan, hydroxyethyl cellulose, hydroxypropyl methyl cellulose, and other high molecular polymers. The content of the thickener in the whitening cosmetic composition is known in the art, and for example, it is generally about 0.1 to 10% by weight based on the total weight of the component (B).
Examples of such preservatives include, but are not limited to, one or more of methylparaben, propylparaben, phenoxyethanol, benzyl alcohol, phenylethyl alcohol, bis (hydroxymethyl) imidazolidinyl urea, potassium sorbate, sodium benzoate, chlorphenesin, sodium dehydroacetate, caprylhydroxamic acid, 1, 2-hexanediol, 1, 2-pentanediol, p-hydroxyacetophenone, caprylyl glycol, glyceryl undecylenate, sorbitan caprylate, ethylhexylglycerin, moutan root extract, and the like. The content of the preservative in the whitening cosmetic composition is known in the art, and for example, it is generally about 0.01 to 2% by weight based on the total weight of the component (B).
The whitening cosmetic composition can be obtained by mixing (a) the yeast fermented birch sap with other cosmetic ingredients according to a method known in the cosmetic industry. For example, it is prepared by using a dissolving tank, an emulsifying pan, a disperser, a transfer pump, etc., which are generally used in the field of cosmetics. The preparation method comprises putting water soluble substance into water phase dissolving kettle, putting oil soluble substance into oil phase dissolving kettle, heating the two kettles to about 80 deg.C, wherein the raw material easy to agglomerate can be pre-dispersed with disperser. After the dissolution is finished, the oil phase and the water phase are conveyed into an emulsifying pot, and homogenized and emulsified for about 5-15 minutes. After emulsification is finished, the temperature of the material body is reduced to normal temperature, optional essence, preservative and the like are added, and the pH of the product is adjusted according to needs. After the relevant detection indexes are qualified, the products can be filled and delivered. The preparation method can be deleted or adjusted according to the requirements of dosage forms.
The whitening cosmetic composition may be formulated into various dosage forms such as solution, suspension, ointment, cream, emulsion, gel, powder, spray, etc. according to need.
Examples
The present invention will be described in further detail with reference to examples. However, it should be understood that these examples and comparative examples are only intended to illustrate the present invention in more detail, and should not be construed as limiting the scope of the appended claims of the present invention in any way.
The yeast strains used in examples 1-4 were from Angel Yeast Inc., strain number ANGEL 1225.
Example 1
(1) Rehydrating active dry yeast
Putting dry yeast into warm water at 35 ℃ for rehydration for 15 minutes according to the proportion of 1.
(2) Preparation of birch juice culture medium
Taking birch juice stock solution (brix 1.12) collected from great Xinggan Ling in northeast as a substrate, adding 2% of cherry powder as a supplementary carbon source, adding 0.1% of monopotassium phosphate and 0.05% of anhydrous magnesium sulfate, and adjusting the pH of a birch juice culture medium to 4.7 by using 1M sodium citrate aqueous solution.
(3) Inoculating fermentation
Adding the birch juice culture medium prepared in the step (2) into a 300L fermentation tank according to the liquid filling amount of 65% (v/v), and sterilizing for 20 minutes at 105 ℃; based on the volume of the fermentation medium, adding a proper amount of yeast seed liquid into the fermentation medium, continuously fermenting for 28 hours at the temperature of 30 ℃, the stirring speed of 350rmp and the ventilation of 1.8vvm, and stopping the fermentation tank to obtain a fermentation liquid product.
(4) Centrifuging to obtain fermentation filtrate and thallus
Processing the fermentation broth product with a butterfly centrifuge, wherein the centrifugation speed is 6000rpm, the centrifugation time is 30 minutes, and the fermented birch sap filtrate as supernatant is obtained and pumped into a storage tank; and obtaining yeast thallus serving as bacterial sludge, and re-suspending the yeast thallus by using supernatant with 5 times of volume to obtain yeast suspension.
(5) Crushing, centrifuging and filtering to obtain soluble yeast lysate
Carrying out high-pressure homogeneous crushing on the yeast suspension obtained in the step (4) by adopting high-pressure crushing equipment under the conditions of 1000 bar and 1.2 liters/min; then, the mixture was filtered using a celite plate to obtain a soluble yeast lysate.
(6) Mixing yeast lysate with fermented birch juice filtrate and filtering
Mixing the yeast lysate obtained in the step (5) with the fermented birch juice filtrate obtained in the step (4), and then filtering the mixture by using a kieselguhr plate frame to obtain a final fermented birch juice filtrate.
The contents of polysaccharide, protein, nucleic acid and total phenol in the filtrate of the fermented birch sap were measured, and the results are shown in table 1.
Example 2
(1) Active dry yeast rehydration
Putting dry yeast into warm water at 35 ℃ for rehydration for 15 minutes according to the proportion of 1.
(2) Preparation of birch juice culture medium
Birch juice stock solution (brix 1.3) collected from Finland was used as a substrate, 1% Roxburgh rose powder was added as a supplementary carbon source, 0.1% monopotassium phosphate and 0.05% anhydrous magnesium sulfate were added, and the pH of the birch juice medium was adjusted to 5.0 using 1M sodium citrate aqueous solution.
(3) Inoculating fermentation
Adding the birch juice culture medium prepared in the step (2) into a 1000L fermentation tank according to the liquid filling amount of 70% (v/v), and sterilizing for 20 minutes at 105 ℃; based on the volume of the fermentation medium, adding a proper amount of yeast seed liquid into the fermentation medium, continuously fermenting for 48 hours at the temperature of 30 ℃, the stirring speed of 350rmp and the ventilation of 1.8vvm, and stopping the fermentation tank to obtain a fermentation liquid product.
(4) Centrifuging to obtain fermentation filtrate and thallus
Processing the fermentation broth product with a butterfly centrifuge, wherein the centrifugation speed is 6000rpm, the centrifugation time is 30 minutes, and the fermented birch sap filtrate as supernatant is obtained and pumped into a storage tank; and obtaining yeast thallus serving as bacterial sludge, and re-suspending the yeast thallus by using supernatant with 4 times of volume to obtain yeast suspension.
(5) Crushing under high pressure, centrifuging, and filtering to obtain soluble yeast lysate
Carrying out high-pressure homogeneous crushing on the yeast suspension obtained in the step (4) by adopting high-pressure crushing equipment under the conditions of 1100 bar and 1.0 liter/min; then using a diatomite plate frame for filtration, obtaining soluble yeast lysate.
(6) Mixing yeast lysate with fermented birch juice filtrate and filtering
Mixing the yeast lysate obtained in step (5) with the fermented birch juice filtrate obtained in step (4), and filtering with a diatomaceous earth plate frame to obtain the final fermented birch juice filtrate product.
The contents of polysaccharide, protein, nucleic acid and total phenol in the fermented birch juice filtrate were measured, and the results are shown in table 1.
Example 3
(1) Active dry yeast rehydration
Putting dry yeast into warm water at 35 ℃ for rehydration for 15 minutes according to the proportion of 1.
(2) Preparation of birch juice culture medium
Concentrated solution (3 times concentrated, brix 3.58) of birch juice from lesser Khingan in northeast is used as substrate, 1% apple powder is added as supplementary carbon source, 0.1% monopotassium phosphate and 0.05% anhydrous magnesium sulfate are added, and 1M sodium citrate aqueous solution is used to adjust pH of birch juice culture medium to 4.8.
(3) Inoculating fermentation
Adding the birch juice culture medium prepared in the step (2) into a 500L fermentation tank according to the liquid filling amount of 70% (v/v), and sterilizing for 20 minutes at 105 ℃; based on the volume of the fermentation medium, adding a proper amount of yeast seed liquid into the fermentation medium, continuously fermenting for 36 hours at the temperature of 30 ℃, the stirring speed of 350rmp and the ventilation of 1.8vvm, and stopping the fermentation tank to obtain a fermentation liquid product.
(4) Centrifuging to obtain fermentation filtrate and thallus
Processing the fermentation broth product with a butterfly centrifuge at 6000rpm for 30 min to obtain fermented birch sap filtrate as supernatant, and pumping into a storage tank; and obtaining yeast thallus serving as bacterial sludge, and re-suspending the yeast thallus by using supernatant with 5 times of volume to obtain yeast suspension.
(5) Crushing under high pressure, centrifuging, and filtering to obtain soluble yeast lysate
Carrying out high-pressure homogeneous crushing on the yeast suspension obtained in the step (4) by adopting high-pressure crushing equipment under the conditions of 1000 bar and 1.3 liters/min; then using a diatomite plate frame for filtration, obtaining soluble yeast lysate.
(6) Mixing yeast lysate and fermented birch juice filtrate, and filtering
Mixing the yeast lysate obtained in step (5) with the fermented birch juice filtrate obtained in step (4), and filtering with a diatomaceous earth plate frame to obtain the final fermented birch juice filtrate product.
The contents of polysaccharide, protein, nucleic acid and total phenol in the fermented birch juice filtrate were measured, and the results are shown in table 1.
Example 4 (1) reconstitution of active Dry Yeast
Putting dry yeast into warm water at 35 ℃ for rehydration for 15-20 minutes according to the proportion of 1.
(2) Preparation of birch juice culture medium
Concentrated solution (concentrated 1.5 times, brix 1.8) of birch juice from lesser Khingan in northeast was used as a substrate, 3% cherry powder was added thereto as a supplementary carbon source, 0.1% monopotassium phosphate and 0.05% anhydrous magnesium sulfate were added, and pH of birch juice medium was adjusted to 5.0 using 1M sodium citrate aqueous solution.
(3) Inoculating fermentation
Adding the birch juice culture medium prepared in the step (2) into a 300L fermentation tank according to the liquid filling amount of 65% (v/v), and sterilizing for 20 minutes at 105 ℃; based on the volume of the fermentation medium, adding a proper amount of yeast seed liquid into the fermentation medium, continuously fermenting for 28 hours at the temperature of 30 ℃, the stirring speed of 350rmp and the ventilation of 1.8vvm, and stopping the fermentation tank to obtain a fermentation liquid product.
(4) Centrifuging to obtain fermentation filtrate and thallus
Processing the fermentation broth product with a butterfly centrifuge at 6000rpm for 30 min to obtain fermented birch sap filtrate as supernatant, and pumping into a storage tank; and obtaining yeast thallus serving as bacterial sludge, and re-suspending the yeast thallus by using supernatant with 5 times of volume to obtain yeast suspension.
(5) High-pressure crushing, centrifugal filtering to obtain soluble yeast lysate
Carrying out high-pressure homogeneous crushing on the yeast suspension obtained in the step (4) by adopting high-pressure crushing equipment under the conditions of 1000 bar and 1.2 liters per minute; then using a diatomite plate frame for filtration, obtaining soluble yeast lysate.
(6) Mixing yeast lysate and fermented birch juice filtrate, and filtering
Mixing the yeast lysate obtained in step (5) with the fermented birch juice filtrate obtained in step (4), and filtering with a diatomaceous earth plate frame to obtain the final fermented birch juice filtrate product.
The contents of polysaccharide, protein, nucleic acid and total phenol in the fermented birch juice filtrate were measured, and the results are shown in table 1.
Example 5
Fermentation was carried out using a single stock solution of birch sap as a medium, and the process conditions were the same as in example 1 except for the medium as a substrate.
Example 6
Fermentation was carried out using a single concentrated birch sap solution of 3 times as a medium, and the process conditions were the same as in example 3 except for the medium used as a substrate.
TABLE 1 Total phenol, polysaccharide, protein, nucleic acid content of the fermented birch sap filtrate product
Example 8: whitening efficacy testing of fermented birch juice filtrate product
In this example, the whitening efficacy of the birch juice filtrate products fermented in examples 1-6 was evaluated based on tyrosinase activity inhibition and melanin inhibition on human melanocytes, as follows.
(1) Evaluation of tyrosinase Activity inhibition Rate
The four centrifuge tubes A, B, C and D were filled with 0.25mL of L-tyrosine (1.5 mM) and 0.25mL of PBS (0.2M, pH 6.8), respectively. An additional 0.25mL of sample solution was added to tubes B and D. Tubes A and C were filled with an additional 0.25mL of PBS solution. Vortex 4 tubes and mix well, after keeping the temperature at 37 ℃ for 10 minutes, add 0.25mL tyrosinase (250U) to tubes C and D, add the same volume of PBS buffer to tubes A and B to make up the volume of the sample. The four tubes were placed at 37 ℃ and the incubation continued for 20 minutes. 200 microliters of the above-mentioned test solution was added to a 96-well plate, and the absorbance at 475nm was measured with a microplate reader. The tyrosine activity inhibition rate calculation formula is as follows:
tyrosinase activity inhibition% = ((C-A) - (D-B))/(C-A) × 100%
The results obtained are shown in table 2 below.
(2) Evaluation of melanin inhibition ratio on human melanocytes
According to 3X 10 5 Inoculating cells to 96-well plates at a cell/well inoculation density, and adding CO at 37 deg.C, 5% 2 95% RH) overnight. When the cell plating rate in the 6-well plate reaches 40-50%, the medicine is administered in groups, the dosage of each hole is 2mL, and each group is provided with 3 multiple holes. CO at 37 ℃ and 5% 2 The incubator was continued for 72 hours. After the completion of the culture, PBS was washed once (1 mL/well), and after washing, 700. Mu.L of 0.25% trypsin was added per well for digestion for 1 minute (37 ℃ C.), the reaction was terminated by immediately adding an equal volume of DMEM containing 10% FBS, the cells were collected into 1.5mL EP tubes after pipetting uniformly, centrifuged at 10000rpm for 10 minutes, and the supernatant was discarded. To each well was added 1.2mL of the mixture (distilled water: absolute ethanol: and diethyl ether =2:5, V/V), vortexed until the liquid was clear, left at room temperature for 30 minutes, centrifuged at 10000rpm for 10 minutes, the supernatant was discarded, 1mL of a 1mol/L aqueous solution of NaOH containing 10% (V/V) DMSO was added, sealed with a sealing film, and left to heat in a water bath at 80 ℃ for 40 minutes. After the heating was completed, the absorbance was measured at a wavelength of 405nm after the temperature had equilibrated to room temperature. The melanin content was determined according to the melanin standard curve. The results obtained are shown in table 2 below.
Table 2: tyrosinase activity inhibition and human cell melanin inhibition of fermented birch juice filtrate product
Example 9: whitening lotion
The fermented birch juice filtrate product prepared in example 1 was used to prepare a whitening lotion, the formulation of which was as follows:
| composition (I) | Content (mass%) |
| Fermented birch juice filtrate product | 95.55 |
| Sodium benzoate | 0.35 |
| 1, 2-pentanediol | 4 |
| Sodium metabisulfite | 0.1％ |
The preparation method of the whitening toning lotion comprises the following steps: mixing the fermented birch juice filtrate product, sodium benzoate, pentanediol and sodium metabisulfite, and filtering.
Using the half-face control test method, 20 volunteers were tested before and 8 weeks after using the product as follows:
1) Measuring the whiteness value L of facial skin by using a CK-skinncolor instrument;
2) Measuring the melanin index of the skin at the same position by adopting a Maxmeter MX18 reflection spectrometer;
3) Facial patterns of the subjects under different light sources at different periods are collected through VISIA-CR, and the skin glossiness, the whiteness value L and the visible stain area of the same measurement area of the face of the subject at different test time points are analyzed by IPP software.
The results show that 8 weeks using the whitening lotion of example 9 can increase the facial skin brightness L by 5.10%, reduce the melanin content by 13.8%, increase the skin brightness by 18.4%, and significantly reduce the area of facial spots in 18 of 20 subjects.
Example 10: whitening face cream
The fermented birch juice filtrate product prepared in example 4 was used to prepare a whitening cream having the following formulation:
| composition (I) | Content (mass%) |
| Fermented birch juice filtrate product | 87.35 |
| Caprylic capric acid triglyceride | 3 |
| Shea butter | 5 |
| Polyglycol ether mixtures | 3 |
| Methylisothiazolinone | 0.1 |
| Amino acid humectant | 3.0 |
| Sodium benzoate | 0.3 |
The whitening cream is prepared as follows: respectively mixing the oil phase component and the water phase component, heating to 75 deg.C, and respectively making into oil phase and water phase; and then adding the obtained oil phase and water phase into a vacuum homogenizer for homogenizing and emulsifying, cooling to 45 ℃, adding sodium benzoate, and stirring uniformly to obtain the product.
Using the half-face control test method, 20 volunteers were tested before and 8 weeks after using the product as follows:
1) Measuring the whiteness value L of facial skin by using a CK-skinncolor instrument;
2) Measuring the melanin index of the skin at the same position by adopting a Maxmeter MX18 reflection spectrometer;
3) Facial patterns of the subjects under different light sources at different periods were collected by VISIA-CR, and the skin gloss, whiteness L and visible stain area of the same measurement area at different test time points on the subjects' faces were analyzed by IPP software.
The results show that 8 weeks using the whitening cream of example 10 can increase the facial skin brightness L by 6.25%, reduce the melanin content by 13.5%, increase the skin brightness by 21.7%, and significantly reduce the area of facial spots in 15 of 20 subjects.
The technical solutions of the above-described embodiments are preferred embodiments of the present invention, and several modifications and changes can be made without departing from the principle of the present invention, and these modifications and changes should also be regarded as being within the protection scope of the present invention.
Claims (12)
1. A method for producing yeast fermented birch sap comprises fermenting in a culture medium containing birch sap and fruit powder with yeast as strain, wherein the content of the birch sap in the culture medium is above 93%, and the content of the fruit powder in the culture medium is 0.5-5% based on the total weight of the culture medium;
wherein the method comprises the steps of:
(1) Adopting saccharomycetes as a strain, and fermenting in a culture medium containing birch juice and fruit powder to obtain a fermentation liquid product;
(2) Centrifuging and filtering the fermentation product to respectively obtain saccharomycete thallus and fermented birch juice filtrate;
(3) Crushing the yeast thalli under high pressure, and then carrying out centrifugal filtration to obtain soluble yeast lysate serving as supernatant; and
(4) Mixing the obtained yeast lysate and the obtained fermented birch juice filtrate, and filtering to obtain a fermented birch juice filtrate product, namely the birch juice fermented by the yeast;
wherein the pH of the medium is 4.5-5.0, and the fruit powder is selected from the group consisting of apple powder, litchi powder, peach powder, cherry powder, roxburgh rose powder, grape powder, and mixtures thereof.
2. The method of claim 1, wherein the birch juice is a concentrated birch juice having a concentration factor of 1.2-10.
3. The method of claim 1 or 2 wherein the yeast is selected from the group consisting of saccharomyces cerevisiae, saccharomyces boulardii, and yarrowia lipolytica.
4. The method according to claim 1 or 2, wherein the content of said birch sap in said culture medium is above 95% based on the total weight of said culture medium.
5. A yeast fermented birch sap obtainable by the process of any one of claims 1 to 4.
6. A yeast-fermented birch sap is obtained by fermenting in a culture medium containing birch sap and fruit powder with yeast as strain, wherein the content of the birch sap in the culture medium is above 93%, and the content of the fruit powder in the culture medium is 0.5-3% based on the total weight of the culture medium;
wherein the pH of the medium is 4.5-5.0;
the birch juice fermented by yeast contains 740-1200mg/L of polyphenol, 6.1-8g/L of polysaccharide, 1.12-1.5% of protein, and 830-1100mg/L of nucleic acid; and
the fruit powder is selected from apple powder, litchi powder, peach powder, cherry powder, roxburgh rose powder, grape powder and mixture thereof.
7. The yeast-fermented birch juice of claim 6, wherein the birch juice is a concentrated birch juice having a concentration factor of 1.2-10.
8. The yeast fermented birch juice of claim 6 or 7 wherein the yeast is selected from the group consisting of saccharomyces cerevisiae, saccharomyces boulardii, and yarrowia lipolytica.
9. The yeast-fermented birch sap as claimed in claim 6 or 7, wherein the content of said birch sap in said culture medium is above 95% based on the total weight of said culture medium.
10. Use of the yeast fermented birch juice according to any one of claims 5-9 in a whitening cosmetic composition.
11. A whitening cosmetic composition comprising the yeast-fermented birch juice of any one of claims 5 to 9.
12. The whitening cosmetic composition of claim 11, wherein the yeast fermented birch sap is present in the whitening cosmetic composition in an amount of more than 0 to less than 100% based on the total weight of the whitening cosmetic composition.
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| CN114632049B (en) * | 2020-12-15 | 2023-10-31 | 浙江养生堂天然药物研究所有限公司 | Hair loss preventing and hair growing composition |
| KR102615522B1 (en) * | 2021-10-01 | 2023-12-20 | 주식회사 코리아나화장품 | Cosmetic composition for moisturizing skin containing fermentative betula alba juice as active ingredient |
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| JP3761082B2 (en) * | 2002-02-12 | 2006-03-29 | 一丸ファルコス株式会社 | Melanin inhibitor or whitening cosmetic composition |
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