JP7466246B1 - Wrinkle improving agent and its manufacturing method - Google Patents

Abstract

[Problem] The objective of the present invention is to provide a new wrinkle-improving agent that is superior to conventional products. [Solution] The wrinkle-improving agent is obtained by adding water to rice, treating the mixture with an enzyme, fermenting it into alcohol, and then treating the liquid portion obtained by solid-liquid separation with a cation exchange resin and recovering the components adsorbed by the cation exchange resin. [Selected Figure] Figure 1

Description

The present invention relates to a wrinkle improving agent and a method for producing the same.

Many topical agents made from rice have been proposed. For example, Patent Documents 1 to 6 disclose, for various uses, compositions containing ground rice as an active ingredient; compositions containing rice extracts as an active ingredient; compositions in which the extract is obtained by adding water or an organic solvent to rice, treating rice with acid or alkali, reacting enzymes or koji with hydrated rice, or compositions obtained by carrying out these procedures with or without heating; and compositions containing active ingredients obtained by reacting enzymes or koji before, simultaneously with, or after rice extraction.

Japanese Patent Application Laid-Open No. 7-252129 Japanese Patent Application Laid-Open No. 7-41426 Japanese Patent Application Laid-Open No. 6-25004 Japanese Patent Application Laid-Open No. 5-339163 Japanese Patent Application Laid-Open No. 5-301823 Japanese Patent Application Laid-Open No. 5-139984

The objective of the present invention is to provide a composition made from rice that is particularly effective in improving wrinkles.

The present invention (1) is a wrinkle improving agent obtained by subjecting a product obtained by adding water to rice or an extract thereof to the action of an enzyme and/or rice koji, or an extract thereof, to alcoholic fermentation and/or lactic acid fermentation to obtain a fermented product, subjecting the fermented product to solid-liquid separation to obtain a liquid portion, passing the liquid portion through a cation exchange resin, and then eluting the components adsorbed to the cation exchange resin.
The present invention (2) relates to the wrinkle improving agent of the above-mentioned invention (1), wherein the fermented product is an alcoholic fermented product.
The present invention (3) provides a method for producing a rice product by adding water to rice or an extract thereof, and then reacting an enzyme and/or rice koji with the resulting product or an extract thereof;
A fermentation step of obtaining a fermented product by alcoholic fermentation and/or lactic acid fermentation after the water addition or extraction step;
A solid-liquid separation step of separating the fermented product into solid and liquid to obtain a liquid portion;
The method for preparing a wrinkle improving agent is characterized by comprising an adsorption/elution step of passing the liquid portion through a cation exchange resin and then eluting the components adsorbed to the cation exchange resin.
The present invention (4) relates to the preparation method according to the invention (3), wherein the fermentation in the fermentation step is alcoholic fermentation.

The present invention provides a composition made from rice that is particularly effective in improving wrinkles.

FIG. 1 shows the results of a collagen production test for the wrinkle improving agents according to the Examples and Comparative Examples. FIG. 2 shows the results of a hyaluronic acid production test for the wrinkle improving agents according to the Examples and Comparative Examples. FIG. 3 shows the results of an NMF production test for the wrinkle improving agents according to the Examples and Comparative Examples.

This embodiment, which is one aspect of the present invention, is preferably a wrinkle improving agent obtained by subjecting rice to water or an extract thereof to the action of enzymes and/or rice koji, or an extract thereof, to alcoholic fermentation and/or lactic acid fermentation to obtain a fermented product, separating the fermented product into solid and liquid to obtain a liquid portion, passing the liquid portion through a cation exchange resin, and then eluting the components adsorbed to the cation exchange resin.

<Production method of wrinkle improvement agent>
The wrinkle improving agent according to the present embodiment can be obtained, for example, by the following steps: (first step) a step of adding water to rice or obtaining an extract thereof, (second step) a step of applying an enzyme and/or rice koji to the product obtained in the first step (or extracting it), (third step) a step of subjecting the product obtained in the second step to alcoholic fermentation and/or lactic acid fermentation to obtain a fermented product, (fourth step) a step of separating the fermented product obtained in the third step from solid to liquid to obtain a liquid portion, and (fifth step) a cation adsorption/elution step of passing the liquid portion obtained in the fourth step through a cation exchange resin and then eluting the components adsorbed to the cation exchange resin. The manufacturing method of the wrinkle improving agent according to the present embodiment will be described in detail below.

<First step>
The first step is a step of adding water to rice or obtaining an extract thereof. Here, "rice" is a concept including not only white rice, brown rice, and germinated rice, but also parts of rice such as white bran and red bran. However, it is preferable to use those that essentially contain a white rice portion, that is, white rice, brown rice, germinated rice, and white bran. Here, white rice and brown rice refer to brown rice and white rice such as non-glutinous rice and mochi rice, regardless of whether they are Japonica or Indica rice, regardless of the variety or type. In addition, white bran and red bran generally refer to 92% or more red bran and 92% or less white bran that are produced during polishing, but a mixture of both may be used. The above raw materials may be subjected to raw material treatment such as soaking, steaming, roasting (including sand roasting, net roasting, hot air roasting, etc.), surface modification such as steaming roasting, freeze-drying, photo-modification such as UV irradiation, pressurized roasting such as pat rice, and frying. Polished rice, brown rice and germinated rice may be used as is or may be crushed before use. Polished rice, brown rice and germinated rice may be crushed into powder by a general method using a grinder or rice polisher.

When producing germinated rice, rice with germ is soaked in water or sprayed with water to germinate. The temperature during germination is 10 to 50°C. However, as long as germination occurs, the temperature and time are not important. If there is a risk of the water spoiling during germination, it is preferable to replace the water or use some kind of preservative to prevent spoilage. Germinated rice here refers to everything from just before germination to when the rice has germinated. This germinated rice is washed thoroughly before use. It may also be dried before use. When extracting rice or subjecting it to enzymatic decomposition or koji treatment, it is more efficient to crush the raw rice into granules or powder because the surface area is larger. It is not necessary to crush the rice, but in this case it takes a long time to decompose and extract the rice tissue.

Next, "extraction" refers to treating rice with a liquid medium. Examples of treatment with a liquid medium include water extraction, acid extraction, alkali extraction, and organic solvent extraction. First, when extracting rice with water, a high extraction temperature is effective, but low temperatures are also sufficient for extraction. However, when the temperature is low, such as 40°C or less, it is desirable to treat the rice so that it does not spoil by making the pH acidic or alkaline, or adding a preservative or alcohol. The extraction time can be long or short as long as the active ingredients can be extracted, and can be determined according to the extraction temperature. Extraction can be performed under pressure, normal pressure, or reduced pressure. In the case of water extraction, the biggest problem is gelatinization. If the rice becomes pasty, not only will the extraction efficiency decrease, but it will also be extremely difficult to carry out in practical use. To prevent this, amylase can be added to cause a reaction, or the rice can be acidified with hydrochloric acid or the like to cut off the starch. This method can be used to fully solve the problem and is completely problem-free in practice. When performing acid decomposition extraction or alkali decomposition extraction, neutralization and desalting are performed as necessary. When extracting with an organic solvent, it is desirable to pulverize or powder the rice as finely as possible before extraction. The organic solvent may be any common organic solvent such as alcohol, acetone, n-hexane, ethyl acetate, or methanol, but it is better to use a safe one, as those that are harmful to the human body must be completely removed after extraction.

<Second step>
The second step is a step of allowing enzymes and/or rice koji to act on (or extract from) the product obtained in the first step (rice plus water or an extract thereof). Here, the enzymes are enzymes that act on rice, such as starch-decomposing enzymes (liquefying enzymes, saccharifying enzymes), proteolytic enzymes, lipolytic enzymes, fibrolytic enzymes, lignin-decomposing enzymes, and pectin-decomposing enzymes. The enzymes may be one type or a combination of two or more types (for example, a combination of liquefying enzymes and saccharifying enzymes). The rice koji is not particularly limited. That is, the type of koji mold and the variety and type of rice are not particularly important. The first and second steps may be performed simultaneously or at different times (that is, enzymatic decomposition and rice koji may be applied before, simultaneously with, or after extraction).

<Third process>
The third step is a step of alcoholic fermentation and/or lactic acid fermentation of the product obtained in the second step. The fermentation form may be either moromi fermentation or liquid fermentation, with moromi fermentation being preferred. These fermentations may be repeated two or more times, or different fermentation methods may be combined. Sugar may be removed by aerobic fermentation using yeast, alcohol precipitation, or the like.

<Fourth step>
The fourth step is a step of obtaining a liquid portion by performing solid-liquid separation on the fermented product obtained in the third step. The method of solid-liquid separation is not particularly limited, and may be, for example, squeezing or filtration.

<Fifth step>
The fifth step is a cation exchange treatment step in which the liquid portion obtained in the fourth step is passed through a cation exchange resin to obtain the components adsorbed on the cation exchange resin. The cation exchange resin is not particularly limited, and may be strongly acidic (sulfo group, etc.) or weakly acidic (carboxyl group, etc.). The method for recovering the components adsorbed on the cation exchange resin is not particularly limited, but is preferably eluted with caustic soda.

<Other processes, etc.>
The manufacturing method of the wrinkle improving agent according to the present embodiment may include steps other than the above-mentioned steps 1 to 5. In particular, the manufacturing method preferably includes a boiling step before and/or after the fifth step (cation exchange treatment step).

<Timing of process implementation, etc.>
The manufacturing method of the wrinkle improving agent according to the present embodiment can basically be carried out in the order of the first step to the fifth step described above. Here, with regard to the first step to the fifth step and other steps described above, the next step may be carried out after the completion of a certain step, the next step may be carried out in parallel before the completion of a certain step, or a certain step and a certain step may be carried out simultaneously. For example, in the above-mentioned first and second steps, when it is assumed that a water extract is obtained by an enzyme, if the enzyme is added after adding water to rice, this corresponds to a case in which the first step (adding water to rice) is followed by the second step (adding enzyme to rice in water), and if rice is added to water containing an enzyme, this corresponds to a case in which the first step and the second step are carried out simultaneously. In addition, the manufacturing method of the wrinkle improving agent according to the present embodiment may include steps other than the above-mentioned first step to the fifth step.

<Properties of anti-wrinkle agents>
<Promotes collagen production>
As described above, the wrinkle improving agent of this embodiment is characterized in that it is obtained by passing a specific liquid portion through a cation exchange resin and then eluting the components adsorbed to the cation exchange resin, and preferably has a collagen production promoting effect of 5% or more, 7.5% or more, 10% or more, 12.5% or more, 15% or more, 17.5% or more, 20% or more, 25% or more, 30% or more, 35% or more, 40% or more (the upper limit is not particularly limited, for example 500%), compared to the specific liquid portion that has not been passed through a cation exchange resin.

<Inhibitory effect on basement membrane degradation>
The wrinkle improving agent according to the present embodiment preferably has an effect of inhibiting basement membrane decomposition. Here, the effect of inhibiting basement membrane decomposition refers to, for example, the effect of inhibiting matrix metalloproteinase (e.g., MMP2).

<Promoting hyaluronic acid production>
As described above, the wrinkle improving agent of this embodiment is characterized in that it is obtained by passing a specific liquid portion through a cation exchange resin and then eluting the components adsorbed to the cation exchange resin, and preferably has a hyaluronic acid production promoting effect of 2.5% or more, 5% or more, 7.5% or more, 10% or more, or 12.5% or more (the upper limit is not particularly limited, for example 500%) compared to the specific liquid portion that has not been passed through a cation exchange resin.

<NMF production promoting effect>
As described above, the wrinkle improving agent of this embodiment is characterized in that it is obtained by passing a specific liquid portion through a cation exchange resin and then eluting the components adsorbed to the cation exchange resin, and preferably has an NMF production promoting effect of 2.5% or more, 7.5% or more, 10% or more, 12.5% or more, 15% or more, 17.5% or more, 20% or more, 22.5% or more, or 25% or more (the upper limit is not particularly limited, for example 500%), compared to the specific liquid portion that has not been passed through a cation exchange resin.

<Applications of wrinkle improvement agents>
The wrinkle improving agent according to the present embodiment is useful as the composition itself or as an added component to external preparations. In particular, it is useful as a wrinkle improving agent having collagen production promoting action, basement membrane decomposition inhibiting action, hyaluronic acid production promoting action, and NMF production promoting action. Here, the wrinkle improving agent according to the present embodiment may contain various additives. For example, the components usually added to medicines, quasi-drugs, and cosmetics (for example, skin cosmetics, bath agents, and detergents), specifically, excipients, binders, disintegrants, lubricants, fragrances, colorants, solubilizing agents, suspending agents, emulsifiers, coating agents, moisturizing agents, detergents, ultraviolet absorbers, and drugs can be mentioned. Specifically, for example, when used as a cosmetic, the composition is adjusted by appropriately blending aqueous components, oily components, powders, surfactants, oils, pH adjusters, preservatives, alcohols, antioxidants, thickeners, pigments, fragrances, animal extracts, plant extracts, and the like that are commonly used in cosmetics as necessary.

(Applicable subjects)
The wrinkle improving agent of this embodiment is not particularly limited to the target, and can be used not only for middle-aged and elderly people, but also for skin disease sufferers and young people (for example, for the purpose of preventing wrinkles as described below).Wrinkles are also not particularly limited, and can be used for small wrinkles (for example, dryness), facial wrinkles, sagging wrinkles (laugh lines), large wrinkles, sagging due to aging, and loss of firmness due to weight loss.In addition, "improvement" as used in this specification is a concept that includes not only the disappearance or reduction of wrinkles, but also the prevention and suppression of wrinkles.

(Method of applying)
The wrinkle improving agent according to the present embodiment is used by applying (e.g., coating, spraying, etc.) to the skin (typically, a transdermal topical agent). For example, it can be used as medicines, quasi-drugs, and cosmetics such as skin topical agents, cleansers, and bath agents. Specifically, it can be provided in various dosage forms such as solubilized systems such as lotions (liquid skin topical agents, skin lotions, etc.), emulsion systems such as emulsions (emulsion foundations, O/W emulsion-type beauty essences, etc.), powders and granules (bath cosmetics, etc.), creams (O/W emulsion-type creams, hand creams, makeup base creams, etc.), ointments (O/W emulsion ointments, etc.), and gas systems (aerosols, etc.). More specifically, in addition to coating, subcutaneous injections, microneedle patches, poultices, and sheet masks can also be mentioned.

(Amount applied)
When the wrinkle improving agent according to the present embodiment is applied to the skin, it is preferable to use a composition containing 0.0001% by mass or more of the wrinkle improving agent based on the total mass of the composition to be applied, more preferable to use a composition containing 0.001% by mass or more, and even more preferable to use a composition containing 0.01% by mass or more. The upper limit is not particularly limited, and is, for example, 100% by mass.

The present invention will be described in more detail below with reference to examples, but the present invention is not limited to these examples.

<Preparation of wrinkle improvement agent>
Example 1
2g of starch decomposition enzyme, 2g of proteolysis enzyme, 2g of lipolysis enzyme, 2g of fiber decomposition enzyme and 1500ml of water were added to 500g of polished rice and left to stand at 50°C for 20 hours. After that, the temperature was gradually raised, boiled and extracted for 5 minutes, filtered, yeast was added and alcohol fermentation was carried out for 20 days. After solid-liquid separation using a filter and heating at 85°C for 30 minutes, the mixture was passed through a cation exchange column (Mitsubishi Chemical DIAION SK104H) and washed with water, and the column adsorbed components were eluted with caustic soda and neutralized to obtain 0.4L.

Example 2
125g of rice koji and 1500ml of water were added to 500g of polished rice, and the mixture was left at 55°C for 48 hours. Yeast was then added, and alcoholic fermentation was carried out for 20 days. The mixture was separated into solid and liquid using a filter, heated at 85°C for 30 minutes, passed through a cation exchange column (Mitsubishi Chemical DIAION SK104H) and washed with water, and the components adsorbed on the column were eluted with caustic soda and neutralized to obtain 0.4L.

Example 3
Brown rice was put into a grinder to obtain 500 g of ground material. 125 g of rice koji and 1500 ml of water were added to the ground material and left at 55°C for 48 hours. Yeast was then added and alcoholic fermentation was carried out for 20 days. After solid-liquid separation using a filter and heating at 85°C for 30 minutes, the mixture was passed through a cation exchange column (Mitsubishi Chemical DIAION SK104H) and washed with water, and the column adsorbed components were eluted with caustic soda and neutralized to obtain 0.4 L.

Example 4
2g of starch decomposition enzyme, 2g of proteolysis enzyme, 2g of lipolysis enzyme, 2g of fiber decomposition enzyme and 1500ml of water were added to 500g of polished rice, and the mixture was left at 50°C for 20 hours. The temperature was then gradually increased, and the mixture was boiled for 5 minutes for extraction, filtered, and then lactic acid bacteria was added and lactic acid fermentation was carried out for 5 days. The mixture was separated into solid and liquid using a filter, heated at 85°C for 30 minutes, passed through a cation exchange column (Mitsubishi Chemical DIAION SK104H) and washed with water, and the components adsorbed on the column were eluted with caustic soda and neutralized to obtain 0.4L.

Example 5
125g of rice koji and 1500ml of water were added to 500g of polished rice, and the mixture was left at 55°C for 48 hours. Yeast was then added, and alcoholic fermentation was carried out for 20 days. The mixture was separated into solid and liquid using a filter, heated at 85°C for 30 minutes, passed through a cation exchange column (Mitsubishi Chemical DIAION SK104H) and washed with water, and the components adsorbed on the column were eluted with 0.2N sodium chloride to obtain 0.4L.

Comparative Example
2g of starch decomposition enzyme, 2g of proteolysis enzyme, 2g of lipolysis enzyme, 2g of fiber decomposition enzyme and 1500ml of water were added to 500g of polished rice and left to stand at 50℃ for 20 hours. After that, the temperature was gradually raised, boiled for 5 minutes, filtered, yeast was added and alcohol fermentation was carried out for 20 days. Solid-liquid separation was carried out using a filter, and the mixture was heated at 85℃ for 30 minutes, and then concentrated under reduced pressure to obtain 0.4L.

<Test Example>
<Test Example 1. Collagen Production Test Evaluation>
(Reagents)
Cells: Human dermis-derived normal fibroblasts (Kuraray)
Culture medium: 10% FBS (Fetal Bovine Serum) + 1% L-glutamine-penicillin-streptomycin-containing DMEM medium (Sigma-Aldrich)
PBS (Sigma-Aldrich)
・L-Ascorbic acid 2-phosphate trisodium ester (FUJIFILM Wako Pure Chemical Industries, Ltd.)
Pepsin (Sigma-Aldrich)
・Acetic acid, special grade (FUJIFILM Wako Pure Chemical Industries, Ltd.)
・Sircol Soluble Collagen Assay Kit (biocolor)
(Test method)
Human dermal fibroblasts were seeded in a 12-well plate for tissue culture at 0.8×10 ⁵ cells/well, and DMEM was added and cultured until confluent. The medium was replaced with DMEM containing 50 μmol/L of trisodium L-ascorbic acid 2-phosphate ester, which was prepared so that the various samples were 1.25%, and cultured for a total of 21 days. All cultures were performed in a 37°C, 5% CO ₂ environment. After the culture was completed, the cells were washed with PBS, and collagen was extracted with a 0.5 mol/L aqueous acetic acid solution containing 1 mg/mL pepsin. The amount of collagen in the extract was determined using a spectrophotometric plate reader according to the commercially available Sircol Soluble Collagen Assay Kit. The test was performed three times independently, and the amount of collagen per well and the relative value when the control was taken as 100% were calculated and compared. Table 1 shows the amount of collagen per well when this test was performed on the rice extracts of Examples 1 to 5 and the Comparative Example. Table 2 shows the relative values of the amount of collagen when this test was performed on the rice extracts of Examples 1 to 5 and the Comparative Example, with the control being taken as 100% (Figure 1 shows the relative values).

Test Example 2: Basement membrane decomposition test
(Reagents)
・MMP enzyme activity measurement kit: MMP-2 Fluorometric Drug Discovery Kit, RED (Enzo)
(Test method)
The enzyme activity was measured by a method specified using a commercially available human MMP enzyme activity measurement kit. The enzyme activity when the control, the positive control NNGH, and the samples diluted to various concentrations were added was measured using a fluorescence detection plate reader under conditions of an excitation wavelength of 535 nm and a detection wavelength of 590 nm, and the relative value was calculated when the control was set as 100%. An enzyme activity curve was created from the sample concentration and the obtained relative value, and the concentration at which the enzyme activity was inhibited by 50%, IC50 (%), was calculated. Table 3 shows the results of in vitro MMP2 enzyme activity measurement for the rice extracts according to Examples 1 to 5 and Comparative Example. The Comparative Example had no enzyme activity inhibitory effect and could not be calculated.

<Test Example 3. Hyaluronic acid production test>
(Reagents)
・Cells: Human 3D epidermal skin model EPI-200 (Kurabo)
Culture medium: EPI-200 dedicated culture medium (Kurabo)
PBS (Sigma-Aldrich)
・Ceramic grinding ball, φ2mm (Toray)
・Quantikine ELISA Hyaluronan (R&D Systems)
・DC Protein Assay (Bio-Rad)
(Test method)
After culturing a human three-dimensional epidermal skin model (φ8 mm) in a dedicated medium for one day, 150 μL of each sample was added to the stratum corneum side, and the culture was cultured for two days while changing the medium and sample every day. All cultures were performed in a 37°C, 5% _CO2 environment. After the culture was completed, the skin model was washed with PBS, removed, PBS was added, and the cells were crushed using a bead crusher. The amount of hyaluronic acid in the extract was determined using an absorption plate reader according to the prescribed test method based on a commercially available dedicated kit. The amount of protein was determined using an absorption plate reader based on a commercially available dedicated kit. The amount of hyaluronic acid was divided by the amount of protein to calculate the amount of hyaluronic acid per unit protein. In addition, a relative value was calculated when the amount of hyaluronic acid per unit protein of the negative control was set to 100%. Table 4 shows the amount of hyaluronic acid per unit protein when this test was performed on the rice extracts according to Examples 1 to 5 and Comparative Example. Table 5 shows the relative values when this test was performed on the rice extracts of Examples 1 to 5 and the Comparative Example, with the amount of hyaluronic acid per unit protein of the negative control taken as 100% (Figure 2 shows the relative values).

<Test Example 4. NMF production test>
(Reagents)
・Cells: Human 3D epidermal skin model EPI-200 (Kurabo)
Culture medium: EPI-200 dedicated culture medium (Kurabo)
Phosphate Buffered Saline (PBS) (Sigma-Aldrich)
・Ceramic grinding ball, φ2mm (Toray)
・RIPA Lysis Buffer system (Cosmobio)
Amino acid mixed standard solution (Shimadzu)
・DC Protein Assay (Bio-Rad)
(Test method)
After culturing a human three-dimensional epidermal skin model (φ8 mm) in a dedicated medium for one day, 150 μL of each sample was added to the stratum corneum side and cultured for two days. All culturing was performed in a 37°C, 5% _CO2 environment. After the end of the culture, the skin model was washed with PBS, removed, PBS was added, and the model was crushed using a bead crusher. The crushing liquid was dissolved in RIPA Lysis Buffer and used for NMF quantification and protein quantification. The protein amount was determined using a spectrophotometric plate reader based on a commercially available dedicated kit. The NMF amount was determined by ortho-phthalaldehyde post-column derivatization using high performance liquid chromatography. The obtained NMF amount was divided by the protein amount to calculate the NMF amount per unit protein. In addition, a relative value was calculated when the NMF amount per unit protein of the negative control was set to 100%. Table 6 shows the NMF amount per unit protein when this test was performed on the rice extracts according to Examples 1 to 5 and Comparative Example. Table 7 shows the relative values when this test was performed on the rice extracts of Examples 1 to 5 and the Comparative Example, with the NMF amount per unit protein of the negative control taken as 100% (Figure 3 shows the relative values).

Claims (6)

A wrinkle improving agent obtained by adding water to rice or an extract thereof, treating the mixture with at least one enzyme selected from amylolytic enzymes, proteolytic enzymes, lipolytic enzymes and fibrolytic enzymes (excluding combinations of amylolytic enzymes, proteolytic enzymes, lipolytic enzymes and fibrolytic enzymes), or with any of the enzymes and rice koji, or an extract thereof, subjecting the mixture to alcoholic fermentation and/or lactic acid fermentation to obtain a fermented product, separating the fermented product into solid and liquid to obtain a liquid portion, passing the liquid portion through a cation exchange resin, and then eluting the components adsorbed to the cation exchange resin. The wrinkle improving agent according to claim 1, wherein the fermented product is alcoholically fermented. The wrinkle improving agent according to claim 1 or 2, wherein the enzyme includes a starch-degrading enzyme. a water addition or extraction step in which at least one enzyme selected from amylolytic enzymes, proteolytic enzymes, lipolytic enzymes and fibrolytic enzymes (excluding a combination of amylolytic enzymes, proteolytic enzymes, lipolytic enzymes and fibrolytic enzymes) or said enzymes and rice koji are allowed to act on the product obtained by adding water to rice or an extract thereof, or an extract thereof is obtained;
A fermentation step of obtaining a fermented product by alcoholic fermentation and/or lactic acid fermentation after the water addition or extraction step;
A solid-liquid separation step of separating the fermented product into solid and liquid to obtain a liquid portion;
and an adsorption/elution step of passing the liquid portion through a cation exchange resin and then eluting the components adsorbed by the cation exchange resin. The preparation method according to claim 4, wherein the fermentation step is alcoholic fermentation. The preparation method according to claim 4 or 5, wherein the enzyme includes an amylolytic enzyme.