KR102196415B1 - Time difference fermentation method of galactomyces containing lichen-derived minerals and cosmetic composition using the same - Google Patents

Abstract

The present invention is fermented by adding a complex containing one or more of minerals (Cu, Zn, Mg, Ca, Se, etc.) derived from lichens in a process of fermentation of galactomyces. More specifically, the present invention relates to: a time difference fermentation method which diversifies the efficacy of fermentation metabolites through time difference fermentation in a process of galactomyces complex fermentation with added lichen-derived minerals to give an effect on whitening, especially through inhibition of tyrosinase activity in addition to existing efficacy; and a cosmetic composition using the same.

Description

Time difference fermentation method of galactomyces containing lichen-derived minerals and cosmetic composition using the same}

The present invention relates to fermentation by adding a complex containing one or more of minerals (Cu, Zn, Mg, Ca, Se, etc.) derived from lichens in the course of galactomyces fermentation, A fermentation method characterized in that it provides an effect on whitening through inhibition of tyrosinase activity in addition to the existing efficacy by diversifying the efficacy of fermentation metabolites through timed fermentation in the process of the added galactomyces complex fermentation. It relates to a cosmetic composition using this.

Minerals, also called inorganic substances, are elements that make up the human body and are not decomposed unlike organic substances that are decomposed when consumed. Minerals that make up the human body are potassium (K), calcium (Ca), selenium (Se), sodium (Na), iodine (I), zinc (Zn), magnesium (Mg), phosphorus (P), sulfur (S), There are chlorine (Cl), copper (Cu), manganese (Mn), iron (Fe), and cobalt (Co), which cannot be synthesized in the body and must be consumed as food. It accounts for about 4% of the body and is involved in various actions such as bone and tooth composition, oxygen transport of blood, and regulation of digestion and osmotic pressure.

The minerals are involved in various metabolic functions of the body to keep the body healthy, but modern people often lack or excessively disproportionate specific minerals. In particular, the cause of itching (pruritus, pruritus), including atopy and dry skin, is suffering from itching related to the skin. The atopic itching is known to be due to an abnormality in the immune system causing histamine to be unnecessarily excreted from the skin, but in most cases it is difficult to clearly identify the cause.However, according to a recent known, it is caused by an imbalance of mineral ions in the cell. It is known that skin-related diseases occur because the skin barrier is damaged and the skin is not moisturized in time and is easily exposed by external bacteria or viruses.

As described above, methods for strengthening the skin barrier to protect skin cells from external bacteria or viruses have been studied.According to the results of the study, natural minerals play a role in resolving skin troubles, and although they cannot directly kill viruses or bacteria, they reproduce. It has a function to prevent it from doing so and make it disappear after a certain period of time, and this is a natural method, so it has been found that there are no side effects. In addition, natural minerals are known to increase skin moisturizing power by activating natural moisturizing factors (NMF), and to prevent skin rejuvenation by activating enzymes (SOD) that remove active oxygen.

A method of using minerals in skin cells as a component of functional cosmetics by eluting minerals from deep sea water, seaweed, and natural minerals is being made.

Meanwhile, linchen is a symbiotic complex of fungi and algae or/and cyanobacteria, and is classified into about 14,000 species worldwide. At one time, it has been proposed to use secondary metabolites of lichens as antimicrobial agents or antiherbivore agents. In addition, some lichen extracts have been used as various treatments in the private sector, and lichen metabolites include antibiotic agents, antimycobacterials, antiviral, analgesic, and antipyretic properties. It has been shown to have a variety of biological activities. Along with this, research results show that minerals extracted from lichens have excellent efficacy in preventing skin aging due to cell ion imbalance.

Meanwhile, a fermented product of a fermented strain was used to improve skin moisturization. Fermented products are known to have a function of activating natural moisturizing factors (NMF) to supply moisture to skin cells and to help form a natural moisturizing barrier, so they are used as cosmetic ingredients for moisture supply.

As a specific example, galactomyces, among fermented strains, is one of the natural yeasts fermented with yeast, and found that the skin texture of artisans who make yeast in a brewery is fine, clear and elastic. Interest in the efficacy and the various active ingredients contained therein has increased. As it is contained in the cosmetic brand SK-II product, it has begun to attract attention, and various vitamins such as vitamins B1, B2, pantothenic acid, B6, folic acid, and various amino acids, which are natural moisturizing factors And rich in various minerals. Kojic acid, which is known for its anti-aging activity of nucleic acids containing more than 7 times that of sardines, is known as a representative whitening ingredient was also known for the first time in the process of fermenting koji into yeast in early 1900. On the other hand, various natural enzymes such as saccharification enzymes and proteases produced during fermentation are known to increase the production and absorption of effective substances, thereby having antioxidant, anti-aging, whitening, and skin moisturizing effects.

Conventionally, functional cosmetics and manufacturing methods using arbutin (Korean registered patent, KR10-038983) have been published for whitening and wrinkle improvement of skin, but arbutin is a natural plant ingredient and has a structure with hydroquinone known as a safe whitening ingredient. It is very similar and has no side effects, but its efficacy is low. In particular, it changes from lipophilic properties to hydrophilicity and changes into a structure that is very difficult to penetrate into the skin, making it difficult to penetrate melanocytes located deep in the skin.

Therefore, the present inventors have made continuous efforts to develop a method for extracting an effective substance having whitening effect of natural ingredients and a cosmetic composition using the same, and as a result, a time difference fermentation method of galactomyces containing lichen-derived minerals and a cosmetic composition using the same In addition to the skin barrier improvement, mineral imbalance, and moisturizing effect, in particular, it was found that there is a whitening effect by inhibiting the activity of tyrosinase, an enzyme that produces melanin pigments by various minerals and kojic acid. Was completed.

An object of the present invention is to provide a whitening effect of skin cells by inhibiting the activity of thiarosinase, a melanin pigment-producing enzyme that causes spots and freckles, in addition to moisturizing, adjusting mineral balance, and reducing inflammation to skin cells. In order to be described below, it is specifically to provide a time difference complex fermentation method of galactomyces to which minerals derived from lichens are added, and a cosmetic composition using the same.

As a means for solving the above problems,

The present invention provides a time difference fermentation method of galactomyces containing minerals derived from lichens.

In addition, culturing the inoculum in the medium (S210); Inoculating a medium in which the inoculum is cultured with galactomuces to perform primary culture (S220); Adding lichen-derived minerals to the primary cultured medium (S230); A first time difference fermentation and separation step (S240) of secondary culture of the lichen-derived mineral-added medium for 12 to 24 hours, and separating 1/3 of the secondary culture (S240); 2 for tertiary culture for an additional 6 to 12 hours (accumulated 18 to 36 hours) of the medium containing the remaining culture from the secondary culture, and to separate 1/2 of the tertiary culture Second time difference fermentation and separation step (S250); The third time difference fermentation in which the culture medium containing the separated and remaining culture of the tertiary culture is further cultivated for an additional 12 hours to 24 hours (accumulated 30 to 60 hours), and the fourth cultured culture is separated Separation step (S260); Mixing and stirring the first to third timed fermentation products separated in the first to third timed fermentation and separation steps (S270); And it provides a time difference fermentation method of the galactomyces (galactomyces) comprising; filtering and discharging the mixed stirred material (S280).

In addition, the inoculum is galactomyces containing any one or more selected from the group including yeast bacteria (Saccharomyces), lactic acid bacteria (Lactobacillus), Bacillus bacteria (Bacillus), yeast (Aspergillus) or photosynthetic bacteria (Rhodobacter). ) Time difference fermentation method is provided.

In addition, the step of inoculating and culturing a medium in which the inoculum is cultured is performed by inoculating the galactomuces in a proportion of 5% to 10% relative to the total proportion of the inoculum of the medium. Provides a method of fermentation with time difference of galactomyces.

In addition, the first time difference fermentation and separation step is a step of separating 1/3 of the secondary culture (S241); Crushing the separated culture (S242); And it provides a time difference fermentation method of galactomyces (galactomyces) comprising; centrifuging and filtering the crushed culture (S243).

In addition, the second time difference fermentation and separation step is a step of separating 1/2 of the tertiary culture (S251); Crushing the separated culture (S252); And it provides a time difference fermentation method of galactomyces (galactomyces) comprising a; centrifuging and filtering the crushed culture (S253).

In addition, the third time difference fermentation and separation step is a step of separating the fourth culture (S261); Crushing the separated culture (S262); And it provides a time difference fermentation method of galactomyces (galactomyces) comprising a; centrifuging and filtering the crushed culture (S263).

In addition, the step of inoculating and culturing a medium in which the inoculum is cultured is performed by inoculating the galactomuces in a proportion of 5% to 10% relative to the total proportion of the inoculum of the medium. Provides a method of fermentation with time difference of galactomyces.

In addition, the step of selecting the lichen-derived mineral lichen (S110); Washing and drying the selected lichens (S120); Crushing the washed and dried lichens (S130); Extracting the mineral complex from the pulverized product (S140); Desalting and filtering the mineral complex (S150); And it provides a time difference fermentation method of galactomyces (galactomyces) comprising a; step of freeze-drying after the desalting and filtering (S160).

In addition, the mineral derived from lichens is Umbilicaria antarctica (Umbilicaria antarctica), Peltigera canina (Peltigera canina), Peltigera praetextata (Peltigera praetextata), Lamarina conduplicans (Ramalina, conduplicans), Ramalina intermedia, Xanthoparmelia farinose, Diplocia canescens, Cetraria aculeate, Cladonia furcata, Schdefe Pseudephebe pubescens, Sphaerophorus globosus, Stereocaulon alpinum, Umbilicaria antarctica, Usnea antarctica , Usnea longissima (Usnea longissima), Usnea barbata (Usnea barbata) and Usnea aurantiacoatra (Usnea aurantiacoatra) or any one or more selected from the group containing Stictanylanderiana (Stictanylanderiana) It provides a time difference fermentation method of galactomyces containing minerals derived from lichens.

In addition, the minerals derived from lichens are from the group containing Na, Fe, K, Mg, Al, Mn, Si, Ca, Zn, I, S, P, Se, Mg, or pharmaceutically or cosmetically acceptable minerals thereof. It provides a time difference fermentation method of galactomyces (galactomyces) containing one or more selected minerals.

In addition, in the step of adding the lichen-derived mineral, galactomyces (galactomyces), characterized in that 0.01% to 0.5% by weight of the lichen-derived mineral is added to the medium obtained by mixing and culturing the inoculum and galactomuces. ) Time difference fermentation method is provided.

In addition, the step of culturing the 2 to 4 provides a time difference fermentation method of galactomyces (galactomyces) characterized in that the culture at 25 ℃ to 36 ℃.

In addition, the washing and drying step provides a time difference fermentation method of galactomyces, characterized in that the lichens are washed with purified water and then dried to have a moisture content of 10% or less.

In addition, the pulverizing step provides a time difference fermentation method of galactomyces (galactomyces), characterized in that pulverized to a size of 350um to 2,000um.

In addition, the step of extracting the mineral complex from the pulverized product includes dispersing the pulverized product at 0.05% to 0.5% by weight relative to purified water, and extracting the pulverized product by stirring at 5rpm to 30 rpm for 1h to 6h at 60°C to 120°C. It provides a time difference fermentation method of galactomyces containing (galactomyces).

In addition, the step of extracting by stirring may include adding one or more organic acids selected from the group including malic acid (malic acid), citric acid (citric acid), fumaric acid, tartaric acid, oxalic acid, propionic acid, lactic acid, etc. (galactomyces) time difference fermentation method is provided.

In addition, the organic acid provides a time difference fermentation method of galactomyces (galactomyces), characterized in that 0.01% to 2.0% by weight compared to the mixture consisting of the purified water and the pulverized product.

In addition, it provides a cosmetic composition using the material produced by the time difference fermentation method of the galactomyces (galactomyces).

The galactomyces complex fermented product obtained by adding the mineral complex derived from lichens of the present invention for time cultivation contains various minerals and kojic acid to inhibit the activity of thiarosinase, an enzyme producing melanin pigment. It prevents skin cells from blemishes and freckles and provides whitening effects.

1 is a flow chart of a time difference fermentation method of galactomyces containing minerals derived from lichens according to an embodiment of the present invention.

Before describing the present invention in detail below, it is understood that the terms used in the present specification are for describing specific embodiments and are not intended to limit the scope of the present invention, which is limited only by the scope of the appended claims. shall. All technical and scientific terms used in the present specification have the same meaning as commonly understood by those of ordinary skill in the art unless otherwise stated.

Throughout this specification and claims, unless otherwise stated, the term "comprise, comprises, comprising" means to include the recited object, step or group of objects, and steps, and any other object It is not used in the sense of excluding a step, a group of objects, or a group of steps.

Meanwhile, various embodiments of the present invention may be combined with any other embodiments unless there is a clear opposite point. Any feature indicated to be particularly desirable or advantageous may be combined with any other feature and features indicated to be desirable or advantageous. Hereinafter, embodiments of the present invention and effects thereof will be described with reference to the accompanying drawings.

The time difference fermentation method of galactomyces containing lichen-derived minerals according to an embodiment of the present invention and a cosmetic composition using the same include a method of extracting minerals from lichens and galactomyces adding lichen-derived minerals. It is composed of a cosmetic composition using a complex fermentation material produced through a method of obtaining a complex fermentation material by culturing the compound with time difference, and the complex fermentation material is effective in alleviating mineral imbalance, moisturizing, and alleviating inflammation, and in particular, various kojic acids (Kojic acid), which inhibits the activity of tyrosinase, a melanin pigment synthase, and provides a whitening effect.

In the time difference complex fermentation method of galactomyces containing minerals derived from lichens according to an embodiment of the present invention and a cosmetic composition using the same, a method of extracting minerals from lichens may be preceded, and minerals from lichens may be extracted. The extraction method may start from the step of selecting lichens (S110).

The time difference fermentation method of galactomyces of the present invention and minerals added to the cosmetic composition using the same are derived from lichens, and the lichens are fungi and algae or/and cyanobacteria. It is defined as a symbiotic complex of.

The lichens of the present invention are lichens that grow naturally in polar or alpine regions, and are Umbilicaria antarctica, Peltigera canina, Peltigera praetextata, and Lamarina condu. Ramalina, conduplicans, Ramalina intermedia, Xanthoparmelia farinose, Diplocia canescens, Cetraria aculeate, Cladonia Pur Cata (Cladonia furcata), Pseudephebe pubescens, Sphaerophorus globosus, Stereocouron alpinum, Umbilicaria antarctica, Uss Includes Usnea antarctica, Usnea longissima, Usnea barbata and Usnea aurantiacoatra or Stictanylanderiana, etc. Any one or more lichens selected from the group

The Umbilicaria antarctica is a lichen collected by researchers from King George Island in Antarctica, and Usnea longissima is a lichen native to Mt. Jirisan, and other lichens are researchers. Lichens collected from the alpine regions of Yunnan, China. These lichens are a kind of lichens that live in an environment unsuitable for general living things such as strong and dry wind, irregular fluctuations in moisture and nutrients, and strong ultraviolet rays throughout the year.

In addition, the minerals derived from lichens added to the galactomyces culture of the present invention refer to inorganic substances, calcium (Ca), phosphorus (P), potassium (K), iron (Fe), zinc (Zn), copper ( Cu), manganese (Mn), selenium (Se), etc. are contained. Minerals are essential for maintaining healthy cells and human body, such as improving the skin health of modern people, especially atopic symptoms and delaying skin aging.

90% of diseases are caused by free radicals, and free radicals in the body promote disease and skin aging, and destroy cells by making lipids, the main component of cell membrane formation, into lipid peroxidation. In the middle-aged, the mineral balance is collapsed and deficient due to a decrease in intake and absorption of minerals, thereby inhibiting the physiological activity of the skin and interfering with the action of the enzyme (SOD) that removes active substances in the body. An essential component of an enzyme that removes active oxygen is iron, copper, zinc, selenium, and manganese contained in lichens.

Even if the protective function of the stratum corneum is damaged, the homeostasis maintenance function that tries to recover it works. In this case, there are research results that the abundant calcium (Ca) and magnesium (Mg) of lichens can effectively improve. In addition, calcium (Ca) promotes the production of intercellular lipids and strengthens keratinocytes, and magnesium (Mg) helps transport intercellular substances to the stratum corneum and helps repair the stratum corneum, and the old keratin is magnesium. (Mg) has the function of increasing the penetration pressure and excreting by combining with the wastes in the pores, so it can remove old dead skin cells.

In addition, zinc (Zn) plays a very important function in the maintenance and regeneration of the skin, and selenium (Se) plays a role as an essential component of glutathione peroxidase, which is a kind of antioxidant enzyme and antioxidant enzyme. It has been reported that it causes anti-inflammatory action by inhibiting the production of kine.

In addition, phosphorus (P) is involved in acid-alkali equilibration in skin cells and buffers skin cell fluids, and when it is deficient, metabolism becomes impaired and it causes poor growth, and potassium (K) is an acid alkali in the body and outside the body. It contributes to equilibrium, maintains the osmotic pressure inside and outside the skin cells to resolve ionic imbalances, and in case of deficiency, it causes aging due to damage to the cell barrier by disrupting skin tissue cells. Iron (Fe), another mineral, binds with oxygen to maintain the metabolism of all tissue cells, and in case of deficiency, it causes glossitis, cleft lip inflammation, spoon-shaped nails and hair loss of the nail, and copper (Cu) It activates melanin-forming enzyme, and when it is deficient, it causes torsion hair, nodular fissure, and bead hair.

In the present invention, "mineral" refers to a mineral extracted from lichens by a conventional method or separated therefrom. In addition, in the present invention, "mineral" refers to a material produced through the extraction process, desalting and filtration process or drying process, and the calcium (Ca), phosphorus (P), potassium (K), iron (Fe), In addition to those referred to as general inorganic substances such as zinc (Zn), copper (Cu), manganese (Mn), and selenium (Se), vitamins and organic substances other than the inorganic substances may be included in the extraction process. “Mineral” can be used interchangeably as “mineral complex”.

After the lichen selection step (S110), a step of washing and drying the selected lichen (S120) may be included, and in the washing and drying step (S120), the selected lichen is washed with purified water and the moisture content is about 10. It can be dried to less than %. If the moisture ?t amount is less than 10%, it is easy to crush lichens in the subsequent crushing step (S130), but if the moisture content is more than 10%, a lot of power is required when crushing lichens due to agglomeration phenomenon. It is inefficient.

After the washing and drying step (S120), a grinding step (S130) may be included. The pulverizing step (S130) is a step of pulverizing the lichen pulverized material to a predetermined size, and may be pulverized to a size of 350 um to 2,000 um with a grinder such as a stainless blender.

After the pulverizing step (S130), a step (S140) of extracting the mineral complex from the pulverized material may be included. In the step of extracting the mineral complex, the pulverized product pulverized to a size of 350 μm to 2,000 μm is dispersed in purified water so that it is 0.05% to 0.5% by weight compared to purified water, and at 60°C to 120°C for 1h to 6h at 5rpm to 30 rpm. It may consist of a step of extracting by stirring.

The reason for dispersing the pulverized product in purified water to be 0.05% to 0.5% by weight compared to purified water is to optimize the extraction of minerals contained in the pulverized product, and if it is less than 0.05% by weight, the content of extracted minerals is too low. There is a problem in that the efficiency is not good because the amount lost in the post-treatment process is large, and when the amount is 0.5% by weight or more, there is a problem in that a significant portion of the mineral contained in the pulverized material cannot be extracted.

In addition, it may consist of a step of extracting by stirring at 5rpm to 30 rpm for 1h to 6h at 60°C to 120°C. If the temperature is below 60°C, the efficiency of the extracted mineral is lowered, and if the temperature is higher than 120°C, grinding Water denatures due to high temperature and extracts unnecessary components other than expected components, which complicates the post-treatment process and increases the cost.If the stirring speed is less than 5 rpm, it is not normally stirred and the pulverized material sinks, resulting in extraction efficiency. When the pulverized material is lowered and is greater than 30 rpm, the pulverized material is pulverized to a smaller size, so that the pulverized material is included in the filtering process in an ultrafiltration apparatus after the extraction process, so that the quality of the final product is not uniform.

Moreover, the extraction step (S140) may consist of extracting the mineral complex by further adding at least one organic acid. If inorganic acids such as hydrochloric acid are used, the extraction efficiency of mineral complexes can be increased, but since there is a problem in that inorganic acids remain in the extract after mineral extraction, organic acids can be used instead of inorganic acids to solve the residual accumulation problem and to secure stability. have.

The organic acid may be composed of one or more organic acids selected from the group including malic acid (malic acid), citric acid (citric acid), fumaric acid, tartaric acid, oxalic acid, propionic acid or lactic acid.

Since the organic acid has a lower mineral extraction efficiency than that of a strong acid, it is important to find an appropriate extraction condition. The organic acid is preferably added in an amount of 0.01% to 2% by weight based on the total mixture of the pulverized product and purified water. If the organic acid concentration is 0.01% by weight to 2% by weight, the yield reaches the highest value, but if it is more than 2% by weight, it is not economical because it shows a constant yield without increasing the yield, and if it is less than 0.01% by weight, the extraction time becomes longer. have. Therefore, it is preferable to appropriately adjust the organic acid concentration within the range of 0.01% to 2% by weight in consideration of the pH conditions according to the application.

After the extraction step (S140), a step (S150) of desalting and filtering the extracted mineral complex may be included.

The desalting step is a step of producing mineral water by removing salts from the mineral complex that has been subjected to the extraction step (S14), and this is to remove salts corresponding to impurities. The desalting step may be performed by a method such as a desalination treatment method by an electric extraction method and a desalination treatment method by an electrodialysis method, and is not limited to a specific method.

The filtration step is a process of removing suspended solid particles from the extract, and may consist of a process of separating using the size of the particles passing through the membrane, and depending on the size of the object to be separated, nanofiltration or microfiltration. , Ultrafiltration (Ultrafiltraion), reverse osmosis (reverse osmosis) methods such as. Considering the size of the mineral particles to be separated (0.05 μm to 1 μm), it is preferable to use an ultrafiltration device in the range of 1 nm to 0.1 μm.

After the desalting and filtering step (S15), a freeze-drying step (S16) may be included. The drying step includes drying methods such as hot air drying, freeze drying, spray drying, and vacuum drying. However, when considering the efficacy of the dried mineral complex, the freeze-dried product is advantageous in terms of efficacy compared to other drying methods. A mineral complex derived from lichen can be obtained using a freeze-drying method.

In the method of manufacturing a complex fermentation material by culturing galactomyces to which minerals derived from lichens are added according to an embodiment of the present invention, it may be configured from the step of culturing the inoculum (S210).

In the inoculum culturing step (S210), the inoculum is any one or more selected from the group containing yeast bacteria (Saccharomyces), lactic acid bacteria (Lactobacillus), Bacillus bacteria (Bacillus), yeast (Aspergillus) or photosynthetic bacteria (Rhodobacter) It can be composed of.

Since the yeast (Saccharomyces) has a very similar cell structure and metabolic rhythm to skin tissue, adding a yeast ingredient as a cosmetic composition and absorbing it into the skin can reduce the cell cycle of the skin to normal with yeast. It is characterized by containing large amounts of amino acids, minerals, and organic acids. Lactobacillus promotes the production of'lolyclean' protein, which accounts for more than 70% of the stratum corneum, and has the function of strengthening the skin barrier by increasing the number of healthy keratinocytes.The pH balance of the skin cells is quickly restored and external bacteria It protects the skin from harmful bacteria growth and has excellent anti-inflammatory effects. Bacillus produces a large amount of butanediol and has a function of holding moisture, so it has the effect of improving skin moisture. Kojic acid of Aspergillus is used as a functional cosmetic composition because it has the effect of releasing spots and contains a large amount of various vitamins, amino acids, minerals, and organic acids. Photosynthetic bacteria (Rhodobacter) are used as cosmetic compositions for treating skin diseases by producing anti-inflammatory substances.

In addition, it may include a step (S220) of inoculating and culturing the galactomyces cultured after the inoculum cultivation step (S210) in the main fermentor. Galctomyces can be inoculated in a proportion of 5% to 10% of the total proportion of the culture medium in the fermentation tank. If Galctomyces is inoculated with less than 5%, there is a possibility that fermentation may not proceed sufficiently due to insufficient amount of inoculum, and if it is added more than 10%, the amount of inoculum is too large and oxygen is Since there is a problem that fermentation does not occur due to insufficient and the culture medium itself is decayed or over-fermented, it is preferable to inoculate with a specific gravity of 5% to 10%.

The galactomyces complex fermentation product is a natural material containing a large amount of organic substances, and can be obtained by fermenting minerals derived from lichens with a galactomices strain.

The fermented product of Galactomyces promotes the conversion of profilaggrin, a precursor of a natural moisturizing factor (NMF), into a natural moisturizing factor, thereby enhancing skin moisturization. In addition, the rich polysaccharide component contained in the fermented product of Galactomyces prevents water evaporation within the skin surface, and is rich in various vitamins such as vitamins B1, B2, pantothenic acid, B6, and folic acid. Various natural enzymes, such as protease, which is a glycation enzyme, increase the production and absorption of effective substances, and are effective in antioxidant, anti-aging, whitening, and skin moisturizing.

In addition, it contains Kojic acid to show the whitening effect of skin cells. Specifically, it refers to 5-hydroxy-2-(hydroxymethyl)-4-pyrone, and the molecular structure can be confirmed through Formula 1 below. Kojic acid is used as a whitening functional cosmetic composition because it has the effect of preventing and treating pigmentation by inhibiting the activity of tyrosinase through copper chelate.

<Formula 1>

In addition, the step of inoculating and culturing galactomyces in the culture medium of the inoculum (S220) is preferably cultured at 34°C to 36°C for 24 to 36 hours. If the cultivation time is less than 24 hours, galactomyces cannot be sufficiently cultured, and if it exceeds 36 hours, galactomyces is overcultured, resulting in insufficient oxygen during the fermentation process, causing rot or physiology. There is a problem in that the fermentation efficiency is lowered due to an increase in the proportion of galactomyces having low activity.

In addition, after the step of inoculating and culturing the galactomyces (S220), a step (S230) of adding the lichen-derived mineral complex obtained through the freeze-drying step (S160). The lichen-derived mineral complex is preferably added in an amount of 0.01% to 0.5% by weight compared to the medium obtained by mixing and culturing the inoculum and galactomyces. If it is less than 0.01% by weight, the effect of the active ingredient of the lichen mineral complex is insignificant, and if it exceeds 0.5% by weight, the excess minerals rather affect the physiological activity of Galactomyces, resulting in lower fermentation efficiency. to be.

In addition, after the step of adding the lichen-derived mineral complex (S230), the medium to which the lichen-derived mineral is added is secondary cultured for 12 to 24 hours, and 1/3 of the secondary culture is separated from the first timed fermentation And it may include a separation step (S240).

Moreover, the first time difference fermentation and separation step (S240) is a step of separating 1/3 of the secondary culture (S241), crushing the separated culture (S242), and centrifuging the crushed culture It may consist of a series of processes of separating and filtering (S243).

In addition, the medium containing the culture remaining after being separated from the secondary culture was further third cultured for 6 hours to 12 hours (accumulated 18 hours to 36 hours), and 1/2 of the third culture was separated It may include a second time difference fermentation and separation step (S250).

Moreover, the second time difference fermentation and separation step (S250) is a step of separating 1/2 of the tertiary culture (S251), crushing the separated culture (S252), and centrifuging the crushed culture And filtering (S253).

In addition, the third time difference in which the culture medium containing the separated and remaining culture from the third culture is further cultured for an additional 12 hours to 24 hours (accumulated 30 to 60 hours), and the fourth culture is separated It may include a fermentation and separation step (S260).

Moreover, the third time difference fermentation and separation step (S260) includes the steps of separating the fourth culture (S261), crushing the separated culture (S262), and centrifuging and filtering the crushed culture. It may consist of a series of processes (S263).

In addition, it may include mixing and stirring the first to third timed fermentation products separated in the first to third timed fermentation and separation steps (S270). However, the mixing ratio between each time difference fermentation is not limited to a specific ratio.

In addition, after the mixing and stirring step (S270) may include a filtering and discharging step (S280). Through the filtering and discharging step, the mixed fermentation material to be obtained in the present invention can be obtained.

In the first to third time difference fermentation and separation steps, when galactomyces containing lichen-derived minerals are fermented at different cumulative times, the complex fermentation material obtained therefrom is between the complex fermentation materials obtained in each time difference fermentation. Since there is a difference in composition and content, additional effects such as tyrosinase inhibitory activity can be obtained compared to the successive fermentation and separation steps.

In another aspect, it relates to a cosmetic composition using a complex fermented product obtained by the manufacturing method of the present invention, wherein the cosmetic composition has an osmotic balance control, inflammation relief, moisturizing effect, and especially a whitening effect. Can be used usefully.

Hereinafter, the content of the present invention will be described in more detail by the following examples. However, it should be noted that the scope of protection of the present invention is not limited by the following examples.

1. Extraction of minerals derived from lichens and time difference fermentation method of galactomyces containing the same

<Example 1>

(1) Lichen-derived mineral extraction method

Usnea barbata species native to Yunnan, China were selected (S110). Thereafter, the selected lichen was washed with purified water and dried so that the moisture content was less than about 10% (S120). Thereafter, the lichen was pulverized into a size of 350um to 2,000um with a stainless blender (S130). Thereafter, purified water was dispersed so that the pulverized product pulverized with a blender was 0.3% by weight relative to the purified water, and malic acid, one of the organic acids, was added so that the ratio of the pulverized product and the purified water mixture was 1% by weight, and at 100°C for 3 hours. The mineral complex was extracted by stirring at 15 rpm (S140). Thereafter, after desalting treatment by the electric extraction method, it was filtered through an ultrafiltration device (Pall, SLP-3053) (S150). Thereafter, lyophilized to obtain a mineral complex derived from lichens (S160)

(2) Time difference fermentation method of galactomyces containing minerals derived from lichens

Saccharomyces 0.3% (v/v) was inoculated and cultured in a medium containing 1.5% (v/v) glucose, 1.5% sugar (v/v), and 1.5% starch syrup (v/v) (S210) . Thereafter, galactomyces was inoculated with a proportion of 10% relative to the proportion of the inoculated bacteria of the medium, followed by primary culture at 35° C. for 30 hours (S220), and then 0.3 compared to the mixed cells of yeast and galactomyces. After adding a lyophilized lichen-derived mineral complex with a weight percent specific gravity (S230), after secondary culture at 30°C for 12 hours, 1/3 of the secondary culture was separated (S241), and the separated culture was crushed Then (S242), the crushed culture was centrifuged and filtered (S243) to obtain a complex fermented product by the first time difference fermentation and separation (S240) step.

Thereafter, the medium containing the culture remaining after being separated from the secondary culture was third cultured at 36° C. for an additional 6 hours (cumulative 18 hours), and then 1/2 of the third culture was separated (S251) , The separated culture was crushed (S252), and the crushed culture was centrifuged and filtered (S253) to obtain a complex fermented product by a second time difference fermentation and separation (S250) step.

Thereafter, the medium containing the culture remaining after being separated from the third culture was cultured at 25° C. for an additional 12 hours (accumulated 30 hours), and then the entire amount of the fourth culture was separated (S261), and The crushed culture was crushed (S262), and the crushed culture was centrifuged and filtered (S263) to obtain a complex fermented product by the third time difference fermentation and separation (S250) step.

Thereafter, the complex fermented product obtained through the 1st to 3rd time difference fermentation and separation process is mixed and stirred in a 1:1:1 ratio (S270), and the mineral galactomyces time difference complex fermentation of the present invention through filtration and discharge steps Water was obtained.

<Example 2>

In Example 1, the second culture time was 24 hours, the third culture time was 12 hours (accumulated 36 hours), and the fourth culture time was changed to 24 hours (60 hours), and was prepared in the same manner as in Example 1. .

<Comparative Example 1>

It was prepared in the same manner as in Example 1, except that the step of extracting the minerals derived from lichens (S110 to S160) and the step of adding the mineral complexes derived from lichens to the fermentation medium (S230) in Example 1 were omitted.

<Comparative Example 2>

In Example 1, the mineral complex derived from lichens was added to the primary culture medium without the first to third time difference fermentation and separation steps (S240 to S260), followed by continuous fermentation at 35°C for 36 hours, and then the fermentation product was 1 to 3 μm. Crushed to size, centrifuged and filtered to obtain a mineral galactomyces complex fermentation product.

<Comparative Example 3>

In Example 1, the second culture time was 28 hours, the third culture time was 16 hours (cumulative 44 hours), except that the fourth culture time was changed to 28 hours (72 hours) was prepared in the same manner as in Example 1. .

2. Mineral analysis of galactomyces complex fermented product containing minerals derived from lichens

Mineral analysis of the galactomyces complex fermentation according to the conditions of Examples 1 to 2 and Comparative Examples 1 to 3 was performed using an inductively coupled plasma mass spectrometer (ICP-MS, X-series (X5, X7), Thermo Elemental, UK). The mineral components and contents were analyzed using, and the analysis results are shown in Table 1 below.

division Ca P K Fe Zn Cu Mn Se Mg Example 1 582 82 62 17 38 84 55 13 41 Example 2 597 93 65 21 42 89 57 15 42 Comparative Example 1 15 5 ND ND ND 3 ND ND ND Comparative Example 2 570 77 48 10 32 76 52 12 34 Comparative Example 3 585 76 59 15 42 81 56 14 42

In Table 1, the units of the constituents are all “ppb”.

As shown in the analysis results in Examples 1 and 2 above, as the second to fourth time difference fermentation time increases, the amount of mineral extraction increases, but as shown in Comparative Example 3, if the time difference fermentation time proceeds longer than necessary, it It can be seen that the amount of extracted or increased is maintained without significant change. This is believed to be due to the fact that galactomyces is overcultured during the second time-difference fermentation and, rather, adversely affects the acquisition of minerals in later stages.

3. Galactomyces Manufacture of cosmetic standard samples containing complex fermented products

A cosmetic standard sample was prepared as shown in Table 2 below by using the galactomyces complex fermented product prepared in Examples and Comparative Examples 1 and 2.

main
Phase Raw material name Formulation Example 1 Formulation Example 2 Formulation Example 3 Formulation Example 4 Formulation Example 5 A Purified water 56.18 56.18 56.18 56.18 56.18 Galactomyces complex fermentation filtrate 10.00 10.00 10.00 10.00 10.00 Glycerine 5.00 5.00 5.00 5.00 5.00 Propanediol 5.00 5.00 5.00 5.00 5.00 Betain 5.00 5.00 5.00 5.00 5.00 Dipropylene Glycol 3.00 3.00 3.00 3.00 3.00 1,2-Hexanediol 0.80 0.80 0.80 0.80 0.80 Disodium EDTA 0.02 0.02 0.02 0.02 0.02 B Xanthan Gum 0.05 0.05 0.05 0.05 0.05 Hydroxyethyl Acrylate/Sodium Acryloyldimethyl Taurate Copolymer 0.40 0.40 0.40 0.40 0.40 Polyacrylate Crosspolymer-6 0.40 0.40 0.40 0.40 0.40 C Polyglyceryl-3 Methylglucose Distearate 2.00 2.00 2.00 2.00 2.00 Cetearyl Alcohol 3.00 3.00 3.00 3.00 3.00 Butyropermum Parkii (Shea) Butter 1.00 1.00 1.00 1.00 1.00 Caprylic/Capric Triglyceride 3.00 3.00 3.00 3.00 3.00 D Methyl Trimethicone 3.00 3.00 3.00 3.00 3.00 Dimethicone 2.00 2.00 2.00 2.00 2.00 E Fragrance 0.15 0.15 0.15 0.15 0.15

In Table 2, the units of the constituents are all "g", and Formulation Examples 1 and 2 are complex fermented products by time difference fermentation of galactomyces containing minerals derived from lichens prepared in Examples 1 and 2, and formulation examples 3 to 5 prepared cosmetic standard samples using the minerals derived from lichens prepared in Comparative Examples 1 to 3, respectively.

4. Cytotoxicity test

Human dermal fibroblast (HDFa) of a cosmetic standard sample prepared from a galactomyces complex fermented product cultured with the addition of lichen-derived minerals of the present invention was mixed with Low Serum Growth Supplement to Medium 106 and penicillin (100 IU/ml). ), in a culture medium containing streptomycin (100 μg/ml) at 37° C., and cultured in an incubator containing 5% CO 2. In order to confirm the toxicity of the prepared cosmetic standard sample, cells were dispensed into a 96 well plate and then cultured for 18 hours under cell culture conditions. After removing the medium and washing with PBS, a new medium and a cosmetic standard sample were added, respectively, and cultured for 18 hours. After 18 hours, in order to measure the viability of the cells, MTT ASSAY was performed to measure the mitochondria activity of living cells to check the viability of cells, and the cell viability was confirmed, and the results are shown in Table 3 below.

division Cell viability (%) Formulation Example 1 118.7 Formulation Example 2 119.2 Formulation Example 3 93.3 Formulation Example 4 109.4 Formulation Example 5 118.9

As shown in Table 3, the fibroblasts showed a high cell survival rate of 100% or more in all samples except 3 in the formulation, and in particular, compared with the cosmetic standard samples according to Formulation Examples 3 and 4, Formulation Example 1 and It can be seen that the cosmetic standard sample of 2 shows a higher cell viability. Formulation Example 5 outside the scope of the present invention also shows a high cell viability, but it can be determined that it is not suitable for commercialization in view of the amount of minerals and cell viability obtained relative to the manufacturing time and cost.

5. Moisturizing power measurement

In order to measure the moisturizing power of the cosmetic standard sample prepared from the galactomyces complex fermented product cultured with the addition of the mineral derived from lichen of the present invention, the moisturizing power was confirmed in Preparation Examples 1 to 5 of the cosmetic standard sample prepared in Table 4 above. This was measured using Corneometer® CM825 (Courage and Khazaka electronic GmbH, Germany).

The skin moisture content was measured before using the cosmetic standard sample, and then the skin moisture content over time was measured again after use. In addition, the skin moisture content when 10 minutes elapsed after using the cosmetic standard sample and the rate of increase in the moisture content of the moisture content before use were measured, and the measurement results are shown in Table 4 below.

division Skin moisture content over time (AU) Skin moisture content increase rate
((After use-Before use)/Before use)) Before use 10 minutes 20 minutes 30 minutes 60 minutes 120 minutes 120 minutes Manufacturing Example 1 27.9 38.3 43.6 46.2 47.3 48.7 74.6% Manufacturing Example 2 28.1 38.8 44.1 47.3 48.1 49.4 75.8% Manufacturing Example 3 28.5 33.9 36.5 38.2 38.9 39.4 38.2% Manufacturing Example 4 28.3 38.6 43.2 46.1 47.2 48.4 71.0% Manufacturing Example 5 28.2 38.7 43.5 46.2 47.5 49.4 75.2%

As a result of measuring the moisturizing power according to the above method, a cosmetic standard sample containing a galactomyces complex fermentation material cultured with a lichen-derived mineral compared to a cosmetic standard sample containing a galactomyces fermentation material cultured without containing a lichen-derived mineral was applied. It was found that the moisture content of one skin was high, and it was confirmed that the case of culturing by the time difference fermentation method was higher than the case of culturing by the continuous fermentation method rather than the time difference fermentation method.

6. Whitening effect-Melanin content analysis

In order to confirm the whitening effect of the galactomyces complex cultured with the addition of lichen-derived minerals of the present invention and the ability to inhibit the production of related melanin, a melanin-inducing substance α-MSH was treated with skin cancer cells and then cultured and compared with the examples. The complex fermented products of Examples 1 and 2 were treated with 0.1 and 0.5%, respectively, and cultured for 3 days to collect cells. After adding 1 mL of 1N NaOH to the collected cells, the cells were completely dissolved by heating, and the absorbance was measured at 475 nm. As a control, arbutin, which is well known as a material having whitening activity, was used at 0.005%, and the measurement results are as shown in Table 5 below.

division density(%) Melanin extraction (%) Example 1 0.1 92.43 0.5 82.34 Example 2 0.1 91.67 0.5 80.17 Comparative Example 1 0.1 156.45 0.5 123.03 Comparative Example 2 0.1 132.24 0.5 107.47 Comparative Example 3 0.1 92.01 0.5 81.63 Arbutin 0.005 57.24

As a result of the melanin extraction analysis by the above method, it was found that the galactomyces complex fermentation material cultured with the minerals derived from lichens was extracted with less melanin compared to the galactomyces complex fermentation material cultured without the minerals derived from lichens. In the case of cultivation by the time difference fermentation method, it was confirmed that melanin was extracted in a smaller amount than when cultured by the continuous fermentation method rather than the time difference fermentation method, indicating that it has excellent whitening activity.

7. whitening effect-hydroxy tea better claim (Tyrosinase) inhibitory activity

In order to confirm the whitening effect of the galactomyces complex fermentation cultured with the addition of lichen-derived minerals of the present invention and the inhibitory effect of related tyrosinase activity, mouse skin cancer cells (B16F10) were added to penicillin (100IU/ml), Streptomycin (100 μg/ml), DMEM (Dulbeco's Modified Eagle's Medium) medium containing 10% FBS was cultured in an incubator containing 5% CO 2 at 37°C.

In order to confirm the tyrosinase inhibitory ability of the galactomyces complex fermentation, the prepared Galactomyces complex fermentation and the melanin-inducing substance α-MSH were treated together in cells, cultured for 3 days, and RNA was extracted to RT. -PCR was performed. The amplified genes were analyzed using the Gel documentation system and Image J program. As a control, arbutin, which is well known as a material having whitening activity, was used at 0.005%, and the measurement results are as shown in Table 6 below.

division density(%) Tyrosinase mRNA
Expression rate (%) Example 1 0.1 94.38 0.5 83.23 Example 2 0.1 93.47 0.5 81.25 Comparative Example 1 0.1 178.45 0.5 173.03 Comparative Example 2 0.1 158.33 0.5 128.97 Comparative Example 3 0.1 93.86 0.5 81.93 Arbutin 0.005 62.24

As a result of analyzing the expression level of tyrosinase mRNA by the above method, the galactomyces complex fermentation material cultured with the minerals derived from lichens was compared to the galactomyces complex fermentation material cultured without containing the minerals derived from lichens. It was found that a small amount of tyrosinase mRNA was extracted, and the case of culturing by the time difference fermentation method had a lower amount of tyrosynase mRNA than the case of culturing by the continuous fermentation method rather than the time difference fermentation method. It was confirmed that it was extracted. It can be seen that the inhibition of melanin by the galactomyces complex fermentation of the present invention is due to the inhibition of tyrosinase enzyme activity, which is a key protein in the melanogenesis reaction.

8. Whitening effect-skin brightness change

In order to check the change in skin brightness of the cosmetic standard sample prepared from the galactomyces complex fermented product cultured by adding the mineral derived from lichen of the present invention, a spectrophotometer (Spectrophotometer CR-2600D, Konica Minolta, Inc. ., Japan) was applied to the subject's facial right cheek area three times in succession, and the average value was calculated and applied to the analysis. The instrument measurement was made before application of the test substance and at 2 and 4 weeks after application, and the measured value of the spectrophotometer consists of three factors, L*, a*, and b*, and L* value is brightness, The a* value represents redness, and the b* value represents yellowness. Therefore, as a value for measuring skin brightness, the results of applying the L* value to the analysis are shown in Table 7 below.

division Before use 2nd week of use 4th week of use Example 1 55.47 58.75 59.46 Example 2 55.35 58.72 59.63 Comparative Example 1 55.45 57.27 58.40 Comparative Example 2 55.46 58.12 58.97 Comparative Example 3 55.42 58.77 59.49

As a result of measuring the change in skin brightness by the above method, it was confirmed that the average value of the L* value in Examples 1 and 2 and Comparative Examples 1 to 3 increased as it progressed to before use, 2 weeks of use, and 4 weeks of use.

However, compared to a cosmetic standard sample prepared with a galactomyces complex fermentation material cultured without containing lichen-derived minerals, the cosmetic standard sample prepared with a galactomyces complex fermentation material cultured with a lichen-derived mineral is the average L* value. Was large, and it was confirmed that the average value of L* value was larger in the case of culturing by the time difference fermentation method than the case of culturing by the continuous fermentation method instead of the time difference fermentation method. Therefore, the present invention is judged to help improve skin brightness.

The features, structures, effects, and the like illustrated in each of the above-described embodiments may be combined or modified for other embodiments by a person having ordinary knowledge in the field to which the embodiments belong. Accordingly, contents related to such combinations and modifications should be interpreted as being included in the scope of the present invention.

Claims (19)

Culturing the inoculum in the medium (S210);
Inoculating a medium in which the inoculum is cultured with galactomuces for primary culture (S220);
Adding lichen-derived minerals to the primary cultured medium (S230);
A first time difference fermentation and separation step (S240) of secondary culture of the lichen-derived mineral-added medium for 12 to 24 hours, and separating 1/3 of the secondary culture;
2 for tertiary culture for an additional 6 to 12 hours (accumulated 18 to 36 hours) of the medium containing the remaining culture from the secondary culture, and to separate 1/2 of the tertiary culture Second time difference fermentation and separation step (S250);
The third time difference fermentation in which the culture medium containing the separated and remaining culture of the tertiary culture is further cultivated for an additional 12 to 24 hours (accumulated 30 to 60 hours) and the fourth cultured culture is separated Separation step (S260);
Mixing and stirring the first to third timed fermentation products separated in the first to third timed fermentation and separation steps (S270); And
Filtering and discharging the mixed agitated material (S280); including, a time difference fermentation method of galactomyces containing minerals derived from lichens.
delete The method of claim 1,
The inoculum is yeast bacteria (Saccharomyces), lactic acid bacteria (Lactobacillus), Bacillus bacteria (Bacillus), yeast (Aspergillus) or photosynthetic bacteria (Rhodobacter) of galactomyces containing one or more selected from the group Time difference fermentation method.
The method of claim 1,
In the step of inoculating and culturing galactomuces in the medium in which the inoculum is cultivated, galacto inoculation of the galactomuces in a proportion of 5% to 10% relative to the total proportion of the inoculating bacteria of the medium The time difference fermentation method of galactomyces.
The method of claim 1,
The first time difference fermentation and separation step is a step of separating 1/3 of the secondary culture (S241);
Crushing the separated culture (S242); And
Centrifuging and filtering the crushed culture (S243); a time difference fermentation method of galactomyces (galactomyces) comprising.
The method of claim 1,
The second time difference fermentation and separation step is a step of separating 1/2 of the tertiary culture (S251);
Crushing the separated culture (S252); And
Centrifuging and filtering the crushed culture (S253); a time difference fermentation method of galactomyces (galactomyces) comprising.
The method of claim 1,
The third time difference fermentation and separation step is a step of separating the fourth culture (S261);
Crushing the separated culture (S262); And
Centrifuging and filtering the crushed culture (S263); a time difference fermentation method of galactomyces (galactomyces) comprising.
The method of claim 1,
In the step of inoculating and culturing galactomuces in the medium in which the inoculum is cultivated, galacto inoculation of the galactomuces in a proportion of 5% to 10% relative to the total proportion of the inoculating bacteria of the medium The time difference fermentation method of galactomyces.
The method of claim 1,
The minerals derived from lichens are
Selecting lichens (S110);
Washing and drying the selected lichens (S120);
Crushing the washed and dried lichens (S130);
Extracting the mineral complex from the pulverized product (S140);
Desalting and filtering the mineral complex (S150); And
The step of freeze-drying after the desalting and filtration (S160); a time difference fermentation method of galactomyces (galactomyces) comprising.
The method of claim 1,
The minerals derived from lichens are Umbilicaria antarctica, Peltigera canina, Peltigera praetextata, Lamarina conduplicans, La Marina. Ramalina intermedia, Xanthoparmelia farinose, Diplocia canescens, Cetraria aculeate, Cladonia furcata, Schdefebe Fube Pseudephebe pubescens, Sphaerophorus globosus, Stereocaulon alpinum, Umbilicaria antarctica, Usnea antarctica, Usnea antarctica From any one or more lichens selected from the group including Usnea longissima, Usnea barbata and Usnea aurantiacoatra or Stictanylanderiana Time difference fermentation method of galactomyces containing derived minerals.
The method of claim 1,
The lichen-derived mineral is one selected from the group containing Na, Fe, K, Mg, Al, Mn, Si, Ca, Zn, I, S, P, Se, Mg or pharmaceutically or cosmetically acceptable minerals thereof Or the time difference fermentation method of galactomyces (galactomyces) containing more minerals.
The method of claim 1,
In the step of adding the lichen-derived mineral, the lichen-derived mineral is added in an amount of 0.01% to 0.5% by weight compared to the medium obtained by mixing and culturing the inoculum and galactomuces. Time difference fermentation method.
The method of claim 1,
The step of culturing the second to fourth time difference fermentation method of galactomyces (galactomyces), characterized in that the culture at 25 ℃ to 36 ℃.
delete The method of claim 9,
The pulverizing step is a time difference fermentation method of galactomyces (galactomyces), characterized in that pulverized to a size of 350um to 2,000um.
The method of claim 9,
Extracting the mineral complex from the pulverized product comprises dispersing the pulverized product in an amount of 0.05% to 0.5% by weight relative to purified water, and extracting the pulverized product by stirring at 5rpm to 30 rpm for 1h to 6h at 60°C to 120°C. Time difference fermentation method of galactomyces.
The method of claim 9,
The step of extracting by stirring is galactomyces (galactomyces) comprising the step of adding one or more organic acids selected from the group containing malic acid (malic acid), citric acid (citric acid), fumaric acid, tartaric acid, oxalic acid, propionic acid, or lactic acid. Time difference fermentation method of.
The method of claim 17,
The organic acid is a time difference fermentation method of galactomyces (galactomyces), characterized in that 0.01% to 2.0% by weight compared to the mixture consisting of the purified water and the pulverized product.
A cosmetic composition using a substance produced by the time difference fermentation method of galactomyces according to any one of claims 1, 3 to 13, and 15 to 18.

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