JP2012126910A - Sphingolipid derived from fermentation lees - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a sphingolipid obtained from fermentation lees, which contains human-type free ceramides or free ceramides, which are highly rare in nature, effective as a component of an external preparation for skin useful for cosmetics or the healing of atopic dermatitis, etc., as a health food, in particular, a humectant, and contains a large amount of the human-type free ceramides.SOLUTION: The sphingolipid is obtained by the extractive purification from the fermentation lees which are by-products in the production of a fermentation product using a malt. The sphingolipid contains a plurality of free ceramides and has a free ceramide content of 5% or greater, further 30% or greater, or, in addition to these conditions, the free ceramide content is 50% of the total sum of the free ceramide content and a coexisting glycosyl ceramide content.

Description

The present invention relates to a fermented koji-derived sphingolipid having a high free ceramide content, which can be used as it is for cosmetics and health foods.
More specifically, it is possible to separate the sphingolipids obtained from plant materials from nature into glucosylceramide and free ceramide by purification, and the former can be used for food and the latter for cosmetics. The present invention relates to a rare free ceramide, particularly a sphingolipid derived from fermented koji with a high human-type free ceramide content.

Japanese traditional fermented foods such as sake, shochu, beer and soy sauce using koji and yeast produce a large amount of fermented koji as a by-product in the production.
For example, in terms of shochu, this fermented rice cake is said to be a by-product of over 600,000 tons of shochu per year, and its disposal is prohibited to dispose of the ocean to prevent marine pollution. Has become a problem.

For this reason, various means have been proposed for effectively using fermented rice cake as a fertilizer or feed.
Among the proposals is, for example, Japanese Patent No. 4071062 (Patent Document 1) in which attention is paid to a functional component in a fermenter and the component is utilized.

The invention described in Patent Document 1 relates to a composition for preventing or treating a tumor that is malignant melanoma.
The composition for the prevention or treatment of tumors that are malignant melanomas is obtained by drying a solid ethanol extract residue obtained by drying a liquid separated from a shochu distiller or a liquid separated from a shochu distiller. It contains an ethanol extract of the obtained solid content.

  In the invention of Patent Document 1, although specific components are not clarified, JP 2007-82427 A (Patent Document 2) clearly shows that glycosylceramide was obtained by extraction from beer koji. The glycosylceramide is effective as a component of health foods and moisturizers.

  Furthermore, JP 2000-80394 A (Patent Document 3) discloses a method for producing sphingolipids in which a sphingolipid is extracted from soy sauce cake with an organic solvent, and the obtained sphingolipid is used as a raw material for cosmetics. It has been shown that

  The sphingolipid has a common structure of a ceramide in which a long-chain fatty acid is an acid amide bond to a sphingoid base, which is a long-chain base, and a glycosphingolipid containing a glycosylceramide in which a sugar is glycosidically bonded to a phosphate and a base. A generic term for various compounds classified as sphingophospholipids.

It has been conventionally known that some of these compounds have physiological activity.
For example, Japanese Patent No. 3068810 (Patent Document 4) shows that a glycosphingolipid having a specific structure extracted from a sponge is effective as an antitumor agent or an immunostimulator.

  Furthermore, JP 2004-131481 A (Patent Document 5) discloses that a glycosylceramide having a specific structure or a salt thereof is an NKT cell activator, an autoimmune disease (for example, systemic lupus erythematosus, systemic scleroderma, It has been shown to be effective as a therapeutic agent for ulcerative colitis, encephalomyelitis, multiple sclerosis, and human type I diabetes, and as a fertility agent.

The sphingolipid, particularly ceramide, also acts as a moisturizing component that prevents water evaporation from the stratum corneum of the skin.
The ceramide constituting the stratum corneum is produced by the degradation of glucosylceramide and sphingomyelin by the action of glucocerebrositase and sphingomyelinase in epidermal cells.

However, the production amount decreases with aging, and with the progress of dryness and wrinkles of the skin with age, and atopic dermatitis, etc., the reduction of sphingolipids in the skin In order to prevent this, as disclosed in Patent Document 3, it is blended in a cream or ointment and used as a skin moisturizer.

Japanese Patent No. 4071062 (Claims) JP 2007-82427 A (Claims, Effects of the Invention) JP 2000-80394 A (Claims) Japanese Patent No. 3068810 (Claims) JP 2004-131481 A (Claims)

In Patent Document 1, shochu distilled spirit contains various components such as fiber derived from cereals, protein, ash, fat, partially unfermented sugar, fermenting microorganisms, organic acids and vitamins. Is disclosed.
Furthermore, a composition for preventing or treating a tumor which is malignant melanoma, containing a liquid ethanol residue or an ethanol extract separated from the shochu distilled spirit is disclosed.
However, there is no disclosure as to whether or not other components are contained in the shochu distiller and what effect the other components have.

In Patent Document 2, a method for efficiently producing a plant ceramide-related substance from beer koji obtained in a production process such as beer, and a plant ceramide-related substance (ceramide, cerebroside) obtained by the method are disclosed. It is disclosed with respect to the product it contains.
However, there is no disclosure about what other substances the plant ceramide-related substance obtained from beer koji contains.

Patent Document 3 discloses that sphingolipids conventionally extracted from nerve tissues such as bovine brain are extracted from soy sauce or soy sauce cake, which is a by-product of soy sauce, with an organic solvent such as hexane.
However, there is no disclosure about specific components of the sphingolipid and what other substances are contained and how they act.

  Further, Patent Document 4 discloses that the glycosphingolipid is extracted from sponges, and Patent Document 5 merely discloses that glycosylceramide is effective as a specific therapeutic agent and abortion agent.

  In view of the present situation, the inventor is a by-product in the production of liquor, shochu, soy sauce, beer, etc., and fermented rice that has been variously studied for its treatment, has a sphingolipid as a functional component, Analysis and identification of sphingolipids as functional components in the fermented koji, in which koji and yeast are essential, because the sphingolipid can be widely applied to various products, medicines, health foods and cosmetics. The research was conducted based on the prediction that the fermented koji could be used more effectively by performing the above.

As a result, the inventor identified the components of the extract from alcoholic solvents of various fermented products. As a result, the fermented product was a by-product in the production of fermented foods using sake, shochu, and soy sauce. It was found that the extract of the alcoholic solvent contained a large amount of sphingolipid and free ceramide.
Furthermore, it has also been found that the content of free ceramide in the sphingolipid can be increased by a simple operation of multiple extractions and purifications.

Furthermore, human-type free ceramide, which is effective as a component of health foods, especially as a moisturizing agent, or a topical skin preparation useful for healing cosmetics or atopic dermatitis, or free ceramide that is extremely rare in nature, and free of human-type The inventors have found that it is possible to provide a sphingolipid containing a large amount of ceramide and containing a large amount of free ceramide, and have completed the present invention.

That is, the invention according to claim 1 of the present invention is
A fermented koji that is extracted and purified from fermented koji, which is a by-product in the production of fermented products using koji, and contains a plurality of free ceramides, and the content of free ceramide is 5% or more. It is a derived sphingolipid.

The invention according to claim 2 of the present invention is
In the sphingolipid derived from fermented koji according to claim 1,
The content of the free ceramide is
It is characterized by being 30% or more.

The invention according to claim 3 of the present invention is
In sphingolipid derived from fermented koji according to claim 1 or claim 2,
The content of the free ceramide is
It is characterized by being 50% or more of the total amount with the coexisting glycosylceramide content.

The invention according to claim 4 of the present invention is
In sphingolipid derived from a fermented rice cake in any one of Claims 1-3,
The free ceramide is
It is characterized by including ceramide X.

The invention according to claim 5 of the present invention is
In the sphingolipid derived from a fermented koji according to any one of claims 1 to 4,
The extraction purification is
Extraction with an alcohol solvent and purification by removing impurities with a non-solvent of free ceramide.

The invention according to claim 6 of the present invention is
In the sphingolipid derived from fermented rice cake according to claim 5,
The alcohol solvent in the extraction is
Methanol or ethanol or their main component, wherein the non-solvent of the free ceramide in the purification is acetone or hexane.

The invention according to claim 7 of the present invention is
In the sphingolipid derived from a fermented koji according to any one of claims 1 to 4,
The purification is
It includes a step of precipitating the extract from the alcoholic solution by adding water to the alcoholic solution in which the extract is dissolved.

The invention according to claim 8 of the present invention provides
In the sphingolipid derived from fermented rice cake according to claim 5,
Purification by the non-solvent is
It is also characterized by being carried out after precipitation of the extract by adding water to the alcoholic solution of the extract.

The invention according to claim 9 of the present invention is
In the sphingolipid derived from fermented rice cake according to claim 5 or 6,
Purification by the non-solvent is
It is characterized by being repeated several times.

The invention according to claim 10 of the present invention provides
In the sphingolipid derived from fermented koji according to any one of claims 5 to 9,
The purification is
The adsorption and removal of impurities in the alcoholic solution of the extract is performed by activated carbon and / or anion exchange resin.

The sphingolipid derived from fermented rice cake of this invention is obtained by extraction and purification of an alcoholic solvent of fermented rice cake, which is a by-product produced in the production of fermented foods using rice cake, and the resulting extract is purified and washed. Further, it is obtained by purification, and the content of free ceramide contains 5% or more, further 30% or more, particularly 50% or more or 90% or more free ceramide. Is something.
In addition, 50% or more of the total amount of the coexisting glycosylceramide and free ceramide is free ceramide, which is a sphingolipid rich in natural free ceramide.

  In the present invention, the fermented koji-derived sphingolipid is used to extract the sphingolipid from the fermented koji using an alcohol solvent, and the resulting extract is washed with a non-solvent of free ceramide to remove impurities, The content of free ceramide is increased by applying simple purification means such as adding water to the alcoholic solution obtained by re-extracting the washed extract with an alcohol-containing solvent. As a sphingolipid, it can be easily obtained.

  In particular, since the fermented koji-derived sphingolipid of the present invention can be obtained using only food additives, it can be obtained using raw foods only for skin application, as a raw material for applications where toxicity is a concern. It is particularly effective.

  Furthermore, it is possible to obtain a large amount of the free ceramide in sphingolipid because it is derived from fermented koji, and since it is mainly human-type or human-type free ceramide, it can be used for food, health It can be used easily and simply as a food ingredient.

  In the present invention, free ceramide refers to J.I. A. It has the same structure or an approximate structure as nine types of human ceramides (ceramide 1 to ceramide 9) introduced by Boustra et al. Together with the skin protecting function.

These human-type ceramides, ceramide 1 to ceramide 9, do not include the optical isomer L-form and the geometric isomer cis-isomer, and are said to have the following properties, respectively. (Biochimica et Biophysica Acta 1758 (2006) 2080-2095).
Therefore, the free ceramide in the fermented koji-derived sphingolipid obtained in the present invention is also considered to have equivalent performance.

Incidentally, ceramide 1 is said to reinforce the binding of intercellular lipids of the multilamellar structure of keratinocytes, and is related to keratosis, ichthyosis, and atopic dermatitis due to deficiency.
Furthermore, the dermatological surgical agent is said to have an effect of improving the elasticity of the skin and the connectivity between cells, and is suitable for aging skin and dry skin.

  Ceramide 2 (stearic acid-sphingosine binding type) has the same structure as ceramide present in the skin and hair, and is a white to gray powder with a specific odor. Suitable for high moisturizing and rough skin.

  Ceramide 3 (stearic acid-phytosphingosine binding type) has the same structure as ceramide present in the skin and is a white to light yellow powder with a specific odor, which restores the barrier function and retains moisture for a long time. Persist. Also used to improve damaged hair.

  Ceramide 4, ceramide 5, and ceramide 6 promote the exfoliating action and smooth the skin.

  Unlike glucosylceramide, which is a lipid that hardly exists in the stratum corneum, these free ceramides are excellent in the moisturizing action and the like when applied because they are present in the skin, that is, human type.

  The sphingolipid of the present invention has a high content of free ceramide, which is extremely rare in nature, and human-type free ceramide, that is, at least 50% or 90% or more of free ceramide in total with glycosylceramide. It can be provided as a sphingolipid derived from fermented rice cake.

  Furthermore, sphingolipids with a high content of free ceramide according to the present invention are used as raw materials for cosmetics or skin external preparations according to the above characteristics and exhibit excellent functions. It is used effectively.

Furthermore, the sphingolipid of this invention contains many human-type free ceramides.
Therefore, even if it is used as it is for cosmetics and health foods, it is expected to exhibit excellent functions, and the glucosyl type and free type are separated and the former is used for food and the latter is used for cosmetics. It is an excellent one that can be used in various ways.

FIG. 1 is a diagram showing the results of silicic acid TLC analysis of soy sauce lees total lipids. FIG. 2 is a diagram showing the results of HPLC analysis of soy sauce koji polar lipids and soybean alkali-stable lipid, where A shows the soy sauce koji analysis results and B shows the soy sauce koji analysis results. FIG. 3 is a diagram showing the structure of ceramide in shochu shochu, soy sauce koji, sake lees koji, and black vinegar moromi koji. FIG. 4 is a diagram showing the results of silicic acid TLC analysis of the soy sauce cake extract and purified product. FIG. 5 is a diagram showing a TLC analysis result of silicic acid for comparison between a soy sauce koji extract purified product and a sake koji koji extract purified product. FIG. 6 shows the structure of the main ceramide present in soy sauce cake.

  In this invention, as a fermented rice cake used for extraction of sphingolipid, a soy sauce cake is mentioned as a preferable thing.

  As is clear from the following examples, other fermented rice cakes, that is, vinegar rice cake, miso, awamori rice cake, etc. are also extracted from shochu, sake sake, black vinegar rice cake, etc. Applicable to the invention, foreign liquors (such as Shaoxing liquor and Makgeolli) can also be applied as long as they use straw.

Extraction of sphingolipids from the fermented rice cake may be performed using an alcohol solvent, particularly ethanol.
A solvent for removing impurities other than free ceramide in the sphingolipid, in other words, a solvent that does not dissolve free ceramide in order to obtain a sphingolipid with a high content of free ceramide, that is, a non-solvent of free ceramide If so, various types are used.
Preferable solvents include hexane and / or acetone.

In addition, there are no special conditions for sphingolipid extraction separation, purification, identification, etc., and normal operations are widely applied.
For example, purification can be performed using column chromatography or the like, but special means are required for mass production.

  Therefore, the alcoholic extract of the fermented koji described in detail in Example 4 below is washed with a non-solvent of free ceramide to remove impurities, and the washed extract is used as an alcoholic solvent. There is a method in which, after dissolving, a step of sequentially adding water to the solution to cause precipitation.

Hereinafter, the fermented koji-derived sphingolipid of the present invention will be described based on specific examples, including its acquisition method.
In addition, the fermented rice cake used is a fermented rice cake made of soy sauce made from soybeans, and a fermented rice cake made of soy sauce made from yellow potato cake and purple potato cake.

<Sphingolipid analysis of soy sauce cake>
300 ml of chloroform / methanol (2: 1, volume ratio) was added to 100 g of pressed rice cake (28% water content), which is a by-product of soy sauce produced by a conventional method, and extracted by stirring at room temperature for 1 hour.
Then, the said extract was filtered and fractionated into the solution and the solid part.
This operation is repeated two more times, and these solutions are combined to form a total extract, and 225 mL of distilled water is added so as to have a ratio of chloroform / methanol / water (8: 4: 3, volume ratio), and the liquid is separated. The mixture was stirred vigorously about 20 times at room temperature using a funnel and allowed to stand.
After confirming that the mixed solution was separated into two layers, the lower layer was concentrated to dryness using a rotary evaporator and weighed. The total lipid amount was 13.4 g.

FIG. 1 shows the result of analysis of total lipids by silicic acid thin layer chromatography (TLC, 0.25 mm, manufactured by Merck & Co., Inc.).
The developing solvent was chloroform / methanol: water (90: 10: 1), and detection was performed by heating after spraying with 50% sulfuric acid.
As shown in FIG. 1, in addition to triglycerides derived from soybean raw materials, this lipid contains a large amount of free fatty acids that are degradation products of lipids.
When the polar lipid fraction was magnified with a dotted line, a spot of free ceramide, which is a lipid hardly present in soybean, was detected in addition to sterol glycoside and glucosylceramide, which are abundant in soybean.

Next, silicic acid column chromatography of the total lipid was performed and fractionated into neutral lipid (NL) and polar lipid fraction (PL).
That is, 100 mg of total lipids were dried and then dissolved in a small amount of chloroform, and subjected to a Sep-Pak silica cartridge (manufactured by Waters).
First, neutral lipid was eluted with 50 mL of chloroform, and then polar lipid was eluted with 30 mL of methanol.
When both fractions were concentrated and dried by a rotary evaporator and weighed, it was found that the total lipid contained 89.5% of the neutral lipid fraction and 10.5% of the polar lipid fraction.

Next, the polar lipid fraction was subjected to HPLC (high performance liquid chromatography) analysis to quantify sphingolipids.
Separation was performed by two-liquid gradient elution, and detection was performed using an evaporative light scattering detector (ELSD, manufactured by Shimadzu Corporation).

<Mobile phase>
Liquid A: hexane / isopropanol / acetic acid (82: 17: 1)
Liquid B: isopropanol / water / acetic acid / triethylamine (85: 14: 1: 0.2)
The analysis result of soy sauce cake is shown in FIG. 2A, and the analysis result of soybean is shown in FIG. 2B.

  Soybean analysis was performed on an alkali-stable lipid fraction obtained by alkaline decomposition of total lipids prepared in the same manner as described above with 0.4 M ethanolic KOH.

  As is clear from FIGS. 2A and 2B, soy sauce cake has a large peak around 4 minutes, which is not present in the original soybean.

  Comparison of retention time with a ceramide standard product (C16 ceramide: Matreya), TLC preparative analysis and GC-MS analysis (detection of sphingoid base) confirmed that this peak was free ceramide. .

  The amount of free ceramide and glucosylceramide determined from the standard curve of standard ceramide and standard glucosylceramide (lower spot from brain brain; Sigma) was 380 mg and 80 mg per 100 g of dry mass as shown in FIG. In proportion, free ceramide was 82.6%.

In addition, the result of the TLC analysis of the purified ceramide obtained from the soy sauce cake is shown in FIG. 4, and the comparison with the free ceramide derived from the sake lees is shown in FIG.

<Sphingolipid analysis of Satsuma shochu>
In the same manner as in Example 1, the lipid analysis of shochu was performed using yellow candy and purple candy.
The raw material used was a solid part (hereinafter referred to as jaundice and purple koji) obtained by separating the shochu waste liquid with a solid-liquid separator.
The lipid content of jaundice (62% moisture) and purple koji (69% moisture) were 2.0 and 1.8 g / 100 g (per fresh mass), respectively.
When the sphingolipid was quantified in the same manner as in Example 1, as shown in FIG. 3, per dry mass, jaundice free ceramide was 204 mg / 100 g, glucosylceramide was 169 mg / 100 g, and purple koji free ceramide was 275 mg. / 100 g, glucosylceramide was 177 mg / 100 g.
The percentage of each free ceramide was 54.7% and 60.8% (FIG. 3).
In addition, the analysis value of the other plant shown in FIG. 3 is an analysis value of Takakuwa et al. (Bioresource Technology, 96, 1089-92, 2005).

Both soy sauce cake and shochu contain a large amount of functional material glucosylceramide (commonly known as plant ceramide), which is already widely recognized in the market.
Takakuwa et al., As shown in FIG. 3, measured the amount of glucosylceramide in various vegetable foods and residues, and reported that there was a little amount in apple pulp.
However, the amount of glucosylceramide in shochu as measured by the inventor is much higher than these, and soy sauce cake is equivalent to apple pulp, but is superior to others.
Furthermore, it should be noted that a large amount of free ceramide, which is hardly contained in raw material soybeans, Satsuma koji, and other natural products, was contained in these fermented koji.

<Ceramide component analysis>
Free ceramide and glucosylceramide obtained from soy sauce cake were separated by silicic acid TLC under the same conditions as described above, and 1 mg each was reacted in 1 ml of 1M methanolic HCL at a temperature of 70 ° C. for 18 hours.
After the reaction, 2 ml of hexane was added, and the extraction by stirring was repeated three times to extract the fatty acid.
The lower layer (methanol layer) was adjusted to pH 10 with 4M KOH, 1 mL of water was added, and the mixture was extracted 3 times with 2 mL of diethyl ether.
The collected diethyl ether layer was washed with distilled water three times, and then the organic layer was dried with nitrogen gas to obtain a sphingoid base fraction.
The obtained sphingoid base fraction was subjected to GC-MS analysis (QP2010, manufactured by Shimadzu Corporation) as a trimethylsilyl derivative.
Column: ULBON HR-1 (50 m, 0.25 mm id, 0.25 μm)
The hexane layer after the previous reaction for 18 hours was dried and methylated with anhydrous 5% methanolic HCL and subjected to GC-MS analysis in the same manner as a fatty acid methyl ester.
The analysis results of fatty acids and sphingoid bases are shown in Table 1.

As shown in Table 1, the fatty acids of glucosylceramide were all α-hydroxy fatty acids.
16h: 0, 22h: 0, and 24h: 0 characteristic of soybean were detected.
Furthermore, 20h: 0 characteristic of wheat and 18h: 0 and 18h: 1 characteristic of Aspergillus were detected, respectively.
Furthermore, it can be seen that the soy sauce cake contains a specific glucosylceramide.

In the notation such as 16h: 0, numbers such as “16” indicate the number of carbon atoms, “h” indicates an α-hydroxyl group, and “last number (such as 0)” indicates the number of double bonds.
The same is true for sphingoid bases, and 9-methyl sphingadienin (d19: 2), which is present only in fungi such as Neisseria gonorrhoeae and yeast, was detected.
The “d” is an abbreviation for dihydroxy base.

The constituent fatty acid of the free ceramide was 85% α-hydroxy fatty acid and the remaining 15% was non-hydroxy fatty acid.
As hydroxy fatty acids, a lot of very long chain fatty acids such as 24 and 22 were detected.
In addition, 2,3 dihydroxy fatty acid (24h ₂ : 0) was also detected, which is considered to be derived from Neisseria gonorrhoeae.
From these analysis results, the main structure of ceramide contained in soy sauce cake is summarized in FIG.

In FIG. 6, the X type is not included in the previous nine types, but was known as a part of the plant and fungal ceramide composition, and was the first to succeed in obtaining a free form. is there.
Since this ceramide X has many hydroxyl groups, it is expected as a material having a high moisturizing effect.

  As apparent from FIG. 6 and Table 1, the free ceramide in the sphingolipid obtained in the present invention is almost trans form, unlike other natural ceramides containing cis isomers. The free ceramide in the fermented sprout-derived sphingolipid obtained in the present invention is regarded as a human ceramide or a ceramide similar to it.

Furthermore, as shown in FIG. 6, the sphingolipid obtained by separation and purification from soy sauce cake and obtained from the results of Table 1 contains four types of free ceramide and three types of glucosylceramide as main components. There was found.
At the same time, it was found that the soy sauce cake contains a large amount of ceramides 6 and 7, and the sake sake cake contains a large amount of ceramide 3, and the human-type free ceramide obtained from each of these soy sauce cakes is judged to have unique characteristics. Is.

<Method for large-scale extraction and separation of free ceramide>
Mass extraction and purification of ceramides contained in soy sauce cake were performed as follows.
3 kg of ethanol was added to 1 kg of soy sauce cake (water 28%), and extraction was performed by stirring at a temperature of 40 ° C. for 3 hours.
The extract composed of the ethanol solution was concentrated to dryness with a rotary evaporator to obtain 95 g of an extract. (Extraction process)
1 L of acetone was added to the obtained extract, washed with stirring, and then cooled and centrifuged at a temperature of 4 ° C.
Acetone 500 mL was again added to the washed extract obtained as a precipitate by the centrifugation, and the mixture was washed twice with stirring for 1 minute and cooling centrifugation twice. (Washing process)
Ethanol (500 mL) was added to the extract obtained as a precipitate, and stirring and centrifugation at a temperature of 40 ° C. for 15 minutes were repeated three times to perform re-extraction. (Re-extraction process)
The re-extracted ethanol solution was decolorized using activated carbon and then an anion exchange resin, and concentrated to dryness. (Impurity removal process)
The re-extract from which impurities were removed obtained by the above-mentioned concentration and drying was dissolved in 100 mL of ethanol and recovered, and an equal volume of water was added for precipitation, followed by cooling and centrifugation at a temperature of 4 ° C. (Precipitation process)
100 mL of acetone was added to the re-extract obtained as a precipitate, and the mixture was stirred and washed for 1 minute, and then cooled and centrifuged for 15 minutes.
To the re-extract obtained as a precipitate, 100 mL of acetone was again added and treated in the same manner. Further, 50 mL of hexane and 50 mL of acetone were added to the precipitate, followed by the same treatment and washing. (Washing process)

The re-extract obtained as a precipitate was dried in a vacuum desiccator for 2 hours. (Drying process)
The dried product thus obtained was ceramide powder (including glucosyl type), and its mass was 4.3 g.

The fermented koji-derived sphingolipid of the present invention contains a large amount of free ceramide, particularly human-type free ceramide, which is rare in nature.
Furthermore, since it is obtained in large quantities from fermented koji, it has been widely used in the industries that handle it as a raw material for health foods, cosmetics, or topical skin preparations, and has a higher utility than ever. There is a possibility that
Furthermore, since it is obtained in large quantities from fermented koji, it has been recognized that it has been obtained from natural products in place of chemical synthetic products that have been used as raw materials for health foods, cosmetics, or external preparations for skin. As a safe new material that cannot be used, it has the potential to be widely used in the industry dealing with health foods, cosmetics, and skin external preparations, as it has more utility than ever.

Claims (10)

A fermented koji that is extracted and purified from fermented koji, which is a by-product in the production of fermented products using koji, and contains a plurality of free ceramides, and the content of free ceramide is 5% or more. Origin sphingolipid. The content of the free ceramide is
The fermented koji-derived sphingolipid according to claim 1, which is 30% or more. The content of the free ceramide is
The fermented koji-derived sphingolipid according to claim 1 or 2, which is 50% or more of the total amount with the coexisting glycosylceramide content. The free ceramide is
The fermented koji-derived sphingolipid according to any one of claims 1 to 3, which contains ceramide X. The extraction purification is
The fermented koji-derived sphingolipid according to any one of claims 1 to 4, which is an extraction with an alcohol-based solvent and a purification in which impurities are removed with a non-solvent of free ceramide. The alcohol solvent in the extraction is
The fermented koji-derived sphingolipid according to claim 5, wherein the main component is methanol or ethanol, or a non-solvent of free ceramide in the purification is acetone or hexane. The purification is
The fermented rice cake-derived origin according to any one of claims 1 to 4, comprising a step of precipitating the extract from the alcoholic solution by adding water to the alcoholic solution in which the extract is dissolved. Sphingolipid. Purification by the non-solvent is
The fermented koji-derived sphingolipid according to claim 5, which is also carried out after precipitation of the extract by adding water to the alcoholic solution of the extract. Purification by the non-solvent is
The fermented koji-derived sphingolipid according to claim 5 or 6, which is repeated a plurality of times. The purification is
10. The fermented rice-derived sphingolipid according to any one of claims 5 to 9, wherein the impurities in the alcoholic solution of the extract are adsorbed and removed by activated carbon and / or an anion exchange resin.