JP2011162491A - Dried extract of malt and oral composition using the same - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a food for preventing and removing stain, and to provide an oral composition. <P>SOLUTION: The dried extract of malt contains a dried product of a mixture containing an extract of the malt and a noncariogenic vehicle. The dried extract of the malt has protease activity and tannase activity. The method for producing the dried extract of the malt includes a step of mixing the malt with water at 0-25°C over a time from 30 min to 3 h, a step of fractionating the extract from the mixture, a step of obtaining a mixture of the extract and the vehicle by mixing the extract with the noncariogenic vehicle, and a step of obtaining the dried extract of the malt by drying the mixture of the extract and the vehicle. <P>COPYRIGHT: (C)2011,JPO&INPIT

Description

  The present invention relates to a food and oral composition for preventing and removing dental stains (also called stains). In particular, the present invention relates to tooth whitening foods and oral compositions having enzyme activity. The present invention further relates to foods using enzymes, foods using rice bran, and oral compositions.

  Stain is dirt that adheres to or deposits on the tooth surface. There are various causes of stains. Examples of endogenous causes of stain include congenital abnormalities, tetracycline, excessive fluoride intake, and aging. Examples of exogenous causes of stain include metals, mouth washes, tobacco, food (tea, wine, etc.) and the like.

  Among them, food-derived stains are tannins (polyphenols) contained in foods such as tea, green tea, coffee, wine, etc. on a film (also called pellicle) made of saliva-derived proteins, polysaccharides, glycoproteins, etc. that covers the tooth surface. It is believed that the dyes such as polyphenols such as polymers), tobacco tar, metal ions, and the like are combined and adsorbed by hydrophobic bonding, oxidative polymerization, and the like.

  Although plaque can be removed by careful brushing with a toothbrush or the like, stains are strongly colored and are difficult to remove by brushing alone. Therefore, it is known that it is different from dental plaque.

To remove colored stains on teeth, there are two targets: pigment and film (pellicle), and conventional methods for removing stains using enzymes can be broadly classified as follows:
(1) a method using an enzyme that decomposes the pigment itself; and (2) a method using an enzyme that decomposes and removes a film (pelicle) covering the tooth surface.

  As an example of the method (1), a method using a tannase (also referred to as a tannin degrading enzyme) (Patent Documents 1 to 4) is known, and as an example of the method (2), a method using a protease (papain). For example, Patent Document 5 for Actinidin, Patent Document 6 for Actinidin, and a method using a lytic enzyme (lysozyme), β-glucanase, mutanase, lipase, dextranase, glucosidase, etc. are known (Patent Document 7).

  Furthermore, apart from the above, oral compositions containing sputum or sputum-derived enzymes are known (Patent Documents 8 to 10).

  All of the prior art are teeth whitening agents using purified enzyme preparations. Enzyme preparations are expensive and cannot be incorporated in large quantities.

  Further, it has not been known that an enzyme relating to pigment degradation and an enzyme relating to pellicle degradation can obtain a higher tooth whitening effect when used in combination than when used alone.

  Although there are compositions for the oral cavity to which an enzyme produced from koji or Aspergillus oryzae is added, there are no compositions containing koji mold or koji itself, and an enzyme purification step is required.

JP 2007-308429 A Japanese translation of PCT publication No. 2003-508589 Special table 2001-509539 gazette Special table 2001-509535 gazette Japanese Patent No. 2628666 JP 2009-515957 A JP 2001-181163 A JP 2006-219441 A JP-A-4-316512 Special table 2009-521508 gazette

  The present invention is intended to solve the above-described problems, and an object thereof is to provide a food and oral composition for preventing and removing stains, and to provide the production thereof.

  As a result of intensive research to solve the above-mentioned problems, the present inventors have used dry cocoon extract produced by a special method so that there is almost no adverse effect on caries and stain prevention effect and stain removal. It discovered that the foodstuff and composition which show an effect were obtained, and completed this invention based on this.

  As a strategy for fighting off diet-derived stain, removal of stain can be considered. In order to remove the stain, the colored stains already attached to the tooth surface are decomposed. That is, the pellicle that is the foundation of the stain is specifically decomposed by the enzyme. Enzymes shown in the following Table 1 can be considered as enzymes that can be used for the degradation of the pellicle.

  None of the enzymes shown in Table 1 degrades enamel.

  On the other hand, regarding the prevention of stains, it is conceivable to decompose a coloring matter that causes coloring before it is attached to a tooth. In this regard, tannase (tannin acyl hydrolase, EC 3.1.1.20, pH 5.0 to 5.5, optimum temperature 40 ° C.) is considered useful. Tannase is an enzyme that specifically degrades the main pigment component tannin in foods and does not degrade enamel. Tannase is used as a food additive to prevent turbidity of tea beverages. This is because the bannin of tea drinks is usually tannin, which is decomposed by tannase to prevent turbidity. Aspergillus oryzae, a mold fungus, can produce tannase.

  That is, in the present invention, as a general picture, by taking daily stain countermeasures in foods containing both enzymes related to stain removal and enzymes related to stain prevention, stain removal and stain prevention are achieved. It can be carried out. As such a prevention mechanism, there is a mechanism that is highly safe for teeth even after continuous use and suitable for daily care.

  The present inventors first considered straw as a candidate raw material. Enzymes produced by mold mold include (1) proteases, amylases, etc. as enzymes that act on pellicle degradation; (2) tannase as enzymes that act on degradation of pigment components.

  Based on the above, the study of sputum as a potential raw material candidate for the removal and prevention of stain revealed that a food and oral composition meeting the purpose of the present invention was produced by producing a dry sputum extract by a special method. It was found that can be manufactured.

  In the present invention, a food material containing both an enzyme involved in pigment degradation (for example, tannase) and an enzyme for degrading the pellicle (for example, protease and amylase) while maintaining the enzyme activity is used. Examples of foods of the present invention include gums, candy (eg, soft candy and caramel) and tablets that are formulated with a dried koji extract.

According to a preferred embodiment of the present invention, the following dry koji extract, food, oral composition and method are provided:
(Item 1) A dried koji extract,
The dry cocoon extract comprises a dry product of a mixture comprising cocoon extract and a non-cariogenic excipient,
The dry koji extract has protease activity and tannase activity.

  (Item 2) The dry candy extract of item 1, wherein the non-cariogenic excipient is selected from the group consisting of sugar alcohols and non-cariogenic saccharides.

  (Item 3) The dry koji extract according to item 2, wherein the total content of sugar alcohol and non-cariogenic sugar in the dry koji extract is 20 wt% to 95 wt%.

  (Item 4) The dry koji extract according to any one of items 1 to 3, wherein the moisture content of the dried koji extract is 0.001 wt% to 5 wt%.

  (Item 5) The dry koji extract according to any one of items 1 to 4, wherein the content of the carious saccharide in the dried koji extract is 20% by weight or less.

  (Item 6) The dried koji extract according to any one of items 1 to 5, wherein the protease activity is 500 U / g or more.

  (Item 7) The dried koji extract according to any one of items 1 to 6, wherein the tannase activity is 50 U / g or more.

  (Item 8) The item 1-7, wherein the non-cariogenic excipient is selected from the group consisting of indigestible dextrin, palatinose, trehalose, lactitol, reduced palatinose, maltitol, erythritol, sorbitol, xylitol, or mannitol. The dry koji extract according to any one of the above.

  (Item 9) The dried rice bran extract according to any one of items 1 to 8, wherein the dried rice bran extract is produced from rice bran.

  (Item 10) A food comprising the dried cocoon extract according to any one of items 1 to 9 and having a carious saccharide content of 0.5% by weight or less.

  (Item 11) The food according to item 10, which is used for removing tongue coating.

  (Item 12) The food according to item 10 or 11, which is a gum, candy or tablet.

  (Item 13) The food according to any one of Items 10 to 12, wherein the protease activity is 100 U / g or more.

  (Item 14) The food according to any one of items 10 to 13, wherein the tannase activity is 10 U / g or more.

  (Item 15) A composition for oral cavity, comprising the dried cocoon extract according to any one of items 1 to 9, wherein the carious saccharide content is 0.5% by weight or less.

  (Item 16) The composition for oral cavity of item 15 which is a dentifrice or a mouthwash.

  (Item 17) The composition for oral cavity according to item 15, which is a troche or gel.

  (Item 18) The composition for oral cavity of any one of the items 15-17 whose protease activity is 50 U / g or more.

  (Item 19) The composition for oral cavity of any one of the items 15-18 whose tannase activity is 5 U / g or more.

(Item 20) A method for producing a dried koji extract according to item 1,
Mixing the straw with water at 0 ° C. to 25 ° C. for 30 minutes to 3 hours;
Separating the extract from the mixture;
A method comprising the steps of mixing the extract and a non-cariogenic excipient to obtain an extract excipient mixture; and drying the extract excipient mixture to obtain a dry anther extract.

  (Item 21) The method according to item 20, wherein the non-cariogenic excipient is a sugar alcohol or a non-cariogenic saccharide.

  (Item 22) The method according to item 20 or 21, wherein the rice bran is a rice bran.

  (Item 23) The method according to any one of items 20 to 22, wherein the drying is freeze drying or spray drying.

  According to the food and oral cavity composition of the present invention, removal of colored stains on the teeth and prevention of deposition can be performed with one food or oral cavity composition. Since the food and oral cavity composition of the present invention have a low content of carious saccharides, the use of the food and oral cavity composition does not adversely affect or hardly affect oral dental caries. The food and oral composition of the present invention can also be effective in removing tongue coating and bad breath in addition to removal of colored stains on the teeth and prevention of deposition.

  The koji extract of the present invention is cheaper and easier to prepare than a plurality of combinations of purified enzymes.

  The food and oral composition using the koji extract of the present invention have higher palatability than the food and oral composition using the enzyme preparation. In addition, since ingredients with experience of eating are used as raw materials, it is safe, can be used continuously on a daily basis, and the effect is enhanced.

  Furthermore, the food and oral composition of the present invention do not require water or tools and can be used easily by anyone.

FIG. 1 shows an outline of a production flow of a dried koji extract from rice koji. FIG. 2 is a graph showing the effect of dry rice bran extract in vitro when hydroxyapatite (Hap) powder is used. FIG. 3 is a photograph showing the effect of dried rice bran extract in vitro when a hydroxyapatite (Hap) disk is used. FIG. 4 is a graph showing the results of the pellicle decomposition effect of Example 1A.

  Hereinafter, the present invention will be described in detail.

(1. Material of dried rice cake extract)
In the present specification, the term “koji extract” refers to an extract obtained by extracting koji with water and / or alcohol.

  In the present specification, “freezing” means that an object (eg, koji extract) solidifies at about 0 ° C. or less. The koji extract can be frozen by methods and equipment known in the art. The koji extract can be frozen, for example, by placing the koji extract in a freezer, or by pouring dry ice, liquid nitrogen, etc. into the koji extract. The final product temperature of the frozen koji extract is preferably about −5 ° C. or lower, more preferably about −10 ° C. or lower, more preferably about −15 ° C. or lower, even more preferably about −20 ° C. or lower, even more Preferably it is about −30 ° C. or less, and still more preferably about −40 ° C. or less. The koji extract is preferably completely solidified within about 3 hours from the start of the freezing treatment, more preferably within about 1 hour, and even more preferably within about 30 minutes.

  In the present specification, “concentration” refers to reducing water in an object (for example, koji extract). The koji extract preferably has a water content in the koji extract after the concentration treatment of about 90 wt% or less, more preferably about 80 wt% or less, based on the weight of the koji extract after the concentration treatment. More preferably about 70% by weight or less, even more preferably about 60% by weight or less, even more preferably about 50% by weight or less, even more preferably about 40% by weight. Most preferably, it is even less preferably about 30 wt%, even more preferably about 20 wt% or less, particularly preferably about 10 wt% or less, Concentrate to about 5 wt% or less. The koji extract can be concentrated using methods and equipment known in the art. During concentration, the koji extract is kept at a temperature of the koji extract of preferably about 50 ° C. or less, more preferably about 40 ° C. or less, and even more preferably about 30 ° C. or less.

  In the present specification, “drying” refers to substantially removing moisture in an object (eg, koji extract). The dried koji extract is preferably about 5 wt% or less, more preferably about 4 wt% or less, more preferably about 3 wt% in the koji extract after the drying treatment based on the weight of the koji extract after the drying treatment. % Or less, even more preferably about 2% by weight or less, even more preferably about 1% by weight or less. The koji extract can be dried using methods and equipment known in the art. During drying, the koji extract is kept at a temperature of the koji extract of preferably about 50 ° C. or less, more preferably about 40 ° C. or less, and even more preferably about 30 ° C. or less.

  In the present specification, “freeze-drying” means that an object (for example, koji extract) is frozen and moisture is directly sublimed in a frozen state under vacuum to substantially dehydrate. The cocoon extract after lyophilization is preferably about 5% by weight or less, more preferably about 4% by weight or less, more preferably about 4% by weight or less in the cocoon extract after lyophilization treatment based on the weight of the cocoon extract after lyophilization treatment. About 3 wt% or less, even more preferably about 2 wt% or less, even more preferably about 1 wt% or less. The koji extract can be lyophilized using methods and equipment known in the art. During freeze-drying, the koji extract is kept at a temperature of the koji extract of preferably about 50 ° C. or less, more preferably about 40 ° C. or less, and even more preferably about 30 ° C. or less.

  The koji extract is preferably treated without impairing the structure of the enzyme contained in the koji extract. In the present invention, the enzyme contained in the koji extract refers to protease, tannase and the like. In producing the dried koji extract of the present invention, it is desirable that the activities of these enzymes are not impaired as much as possible.

(1.1 麹)
In the present specification, the term “麹” refers to a product obtained by breeding Aspergillus on a grain. In this specification, the term “rice bran” refers to white rice grown with Aspergillus oryzae. On November 22, 1989 (Heisei 1), National Tax Agency Notification No. 8 “Making Sake Sake Quality Labeling Standards,” “Koji Koji is made by breeding Koji fungi on white rice. It means that the starch can be saccharified, and the sake with a specific name means that the percentage of koji rice used (the ratio of the weight of koji rice to the weight of white rice, the same shall apply hereinafter) is limited to 15% or more. ”.”

(1.1.1 Aspergillus oryzae)
In the present specification, the term “moji mold” refers to a mold conventionally used for producing koji. Aspergillus oryzae is preferably a mold that can be propagated on a cereal and used as a cocoon among Aspergillus, Monascus, Rhizopus and Mucor. The Aspergillus oryzae that can be used in the present invention is preferably an edible Aspergillus oryzae that can secrete a substance having proteolytic activity and a substance having tannin degrading activity. The Aspergillus oryzae that can be used in the present invention is preferably Aspergillus oryzae that has been conventionally used for preparing koji. Aspergillus is a natural resident fungus. Aspergillus oryzae has a strong property of degrading starch into glucose and protein into amino acids, and depending on the species, it effectively decomposes and absorbs fat. Yes. In the production of fermented food, conventionally, koji mold is propagated in rice, rice bran, wheat, soybean, etc., and subcultured. This is called “Tane Koji” and is used for making koji.

  The seed varieties according to the shape are classified into granular seed potatoes and powdery seed potatoes. The granular seed rice refers to dried rice with a large amount of gonococcal spores. The powdery seed meal refers to a product obtained by collecting only spores from the above.

  Examples of Aspergillus oryzae that can be used in the present invention include: oryzae, A.M. sojae, A.M. Tamarii, A.M. awamori, A.M. Kawachii, A.K. Saitoi, A.I. glaucus, A.M. niger, A.M. Japanics, Monascus purpureru, Monascus anka, Rhizopus and Mucor. Aspergillus used in the present invention may be any commercially available Aspergillus. Aspergillus oryzae used in the present invention is preferably A. oryzae or A.I. Kawachii, most preferably A. oryzae.

  These Aspergillus are sometimes referred to by different names depending on the color of the spores. For example, A.I. oryzae, A.M. sojae, and A.I. Tamarii is called yellow gonococcus because it forms yellowish green spores as the spore infestation progresses. Yellow koji molds are used in the production of miso, soy sauce and sake, and in recent years, some manufacturers use it for shochu. In addition, at the stage where it is used as koji in the production process of ordinary brewed foods, the koji appearance is white, and it is rare to proceed with spore growth until yellow or green can be clearly confirmed. Among the yellow koji molds, there is an albino individual called A. white. There is also oryzae. A. Some kawachii have a brown color as spore growth progresses, and are also called white mold. In addition, A. Awamori forms black-brown spores and is also referred to as black koji mold. Black koji mold is a bacterial species that has been conventionally used for the production of Awamori in Okinawa. In recent years, it is used not only for shochu but also for various foods due to its functionality. Black koji molds are characterized by producing citric acid, which is useful for preventing contamination of bacteria during charging in warm regions. Various seed potatoes are produced and sold by manufacturers specializing in seeds.

(1.1.2 Grain)
As a grain raw material used for the production of koji, any grain capable of breeding Aspergillus can be used. Examples of cereal raw materials used in the production of koji include rice, wheat (eg, wheat, barley, rye, etc.), beans (eg, soybeans, mung beans, empty beans, red beans, etc.), buckwheat and the like. It is preferable to use rice as the grain. The grain may be unmilled (for example, brown rice), may be milled, or may be bran or bran obtained by milling. For example, when rice is used as a raw material for grains, the degree of whitening of rice may be about 70 or more, about 80% or more, or about 90% or more, and the degree of whitening is about 91% or less, about 85% or less, or about 75% or less. And so on. For example, when barley is used as a grain raw material, the degree of whitening of barley can be about 85-88%.

(1.1.3 Method for producing straw)
The cocoon can be produced by methods known in the art. For example, a grain (eg, rice) is immersed in water, drained or steamed as it is, then cooled to a predetermined temperature (eg, about 30 ° C. to about 60 ° C.), and then koji mold is mixed. Aspergillus grows on the cereal by incubating the mixture at a predetermined temperature (eg, about 30 ° C. to about 60 ° C.).

  The incubation temperature is preferably about 20 ° C. or higher, more preferably about 25 ° C. or higher, particularly preferably about 30 ° C. or higher, and most preferably about 35 ° C. or higher. The incubation temperature is preferably about 70 ° C. or less, more preferably about 65 ° C. or less, particularly preferably about 60 ° C. or less, and most preferably about 55 ° C. or less. Of course, the incubation temperature can be adjusted according to the optimum culture conditions for Aspergillus oryzae.

  Furthermore, you may change the temperature to incubate by the stage which propagates Aspergillus oryzae on a grain, and the stage which secretes enzymes, such as protease, from the propagated Aspergillus.

  The incubation time can be adjusted appropriately depending on the condition of Aspergillus on the grain. For example, when the cereal is steamed rice, the incubation time is preferably about 3 hours or more, more preferably about 5 hours or more, still more preferably about 10 hours or more, and particularly preferably about 15 hours or more. Most preferably, it is about 20 hours or longer. For example, when the cereal is steamed rice, the incubation time is preferably about 72 hours or less, more preferably about 60 hours or less, even more preferably about 48 hours or less, and particularly preferably about 46 hours or less. Most preferably, it is about 44 hours or less.

  For example, when the cereal is steamed beans, the incubation time is preferably about 3 hours or more, more preferably about 5 hours or more, still more preferably about 10 hours or more, and particularly preferably about 15 hours or more. Most preferably, it is about 20 hours or longer. For example, when the cereal is steamed beans, the incubation time is preferably about 5 days or less, more preferably about 4 days or less, even more preferably about 3 days or less, and particularly preferably about 72 hours or less. Most preferably, it is about 60 hours or less.

  For example, when the grain is steamed wheat, the incubation time is preferably about 3 hours or more, more preferably about 5 hours or more, still more preferably about 10 hours or more, and particularly preferably about 15 hours or more. Most preferably, it is about 20 hours or longer. For example, when the cereal is steamed wheat, the incubation time is preferably about 5 days or less, more preferably about 4 days or less, even more preferably about 3 days or less, and particularly preferably about 72 hours or less. Most preferably, it is about 64 hours or less.

  Table 2 below summarizes the typical conventional manufacturing methods for straw. In the present invention, any scissors produced by a conventional manufacturing method can be used.

(1.2 Excipient)
In order to produce a dried koji extract, it is difficult to dry the koji extract as it is, and therefore it is preferable to use an excipient in order to promote drying and stabilize the quality after drying. The excipient is preferably a non-cariogenic substance, and more preferably a non-cariogenic carbohydrate. Excipients that can be used for the production of the dried koji extract are preferably non-cariogenic sugars or sugar alcohols. Examples of non-cariogenic saccharides that can be used include indigestible dextrin, palatinose and trehalose. Examples of sugar alcohols that can be used include reduced palatinose (also called palatinit), lactitol, maltitol, erythritol, sorbitol, xylitol, mannitol and the like. The excipient is preferably selected from the group consisting of indigestible dextrin, palatinose, trehalose, lactitol, reduced palatinose, maltitol, erythritol, sorbitol, xylitol and mannitol. These excipients are non-cariogenic substances, and even when eaten, they do not produce acid in the oral cavity and do not cause caries.

Indigestible dextrin is a water-soluble dietary fiber and is also called dextrin-rich dextrin. The indigestible dextrin that can be used in the present invention can be any indigestible dextrin available in the art. For example, the indigestible dextrin can be prepared by hydrolyzing a heat-treated starch with amylase, separating the indigestible component from the undegraded product, and desalting and decolorizing. The indigestible dextrin may be prepared, for example, according to the methods described in JP-A-2-145169, JP-A-2-154664, and the like. The indigestible dextrin may also be pine fiber, fiber sol or the like manufactured by Matsutani Chemical Industry Co., Ltd. (The molecular weight of indigestible dextrin is considered to be unstable, so it was deleted)
For example, the taste of lactitol is sweet but not off-taste.

  The taste of reduced palatinose is similar to sucrose and has no off-flavor (abnormal taste). Further, reduced palatinose has a feature that even if candy is produced using only reduced palatinose as an excipient, it becomes a non-sticky candy.

  The taste of palatinose is similar to sucrose and has no off-taste. In addition, palatinose has the following characteristics: it is non-cariogenic; it is gently digested and absorbed in the body to become energy, and does not change blood glucose level or insulin secretion abruptly; acid-resistant and low-sweetness with excellent taste quality It is a material.

  The taste of maltitol is slightly “blurred” compared to sucrose and has a slight taste that stimulates the throat. Maltitol is as non-hygroscopic as sucrose.

  Erythritol has a unique sweetness and develops sweetness quickly. Since erythritol has an extremely high endothermic effect when dissolved (-43 cal / g), it has a great cool feeling. Erythritol also has a feature that it is less susceptible to diarrhea than other sugar alcohols.

  The taste of sorbitol is "blurred" compared to sucrose and has a slightly irritating taste. Sorbitol is excellent in permeability, moisture retention and shelf life.

  Xylitol has a unique sweetness. Xylitol also has a great cool feeling due to its extremely high endothermic effect upon dissolution (-37 cal / g). Xylitol is evaluated to have a better taste than erythritol. Xylitol is also more effective in promoting saliva secretion than other sugar alcohols.

  It is preferable not to use sugars such as sucrose, glucose, and lactose that are usually used as excipients. This is because when these saccharides are eaten, acid is generated in the oral cavity and the pH in the oral cavity is lowered to cause caries.

(1.3 Other materials)
In the production of the dried cocoon extract, any material commonly used in pulverization can be used as long as it does not promote stain and caries. The dry cocoon extract of the present invention may contain other components useful for the stain removal action or the stain prevention action as necessary as long as they do not adversely affect the stain removal action, the stain prevention action and the caries prevention. Examples of other components useful for stain removal or stain prevention include organic acids.

(2. Method for producing dried koji extract)
The method for producing a dried koji extract of the present invention comprises a step of mixing the koji with water at 0 ° C. to 25 ° C. for 30 minutes to 3 hours; a step of separating an extract from the mixture; Mixing with an excipient (eg, sugar alcohol or non-cariogenic saccharide) to obtain an extract excipient mixture (eg, extract sugar-alcohol mixture or extract non-cariogenic saccharide mixture); and the extraction Drying the liquid excipient mixture (eg, extract liquid sugar alcohol mixture or extract non-cariogenic saccharide mixture) to obtain a dry koji extract may be included.

  The method for producing the dried koji extract of the present invention will be described in more detail below.

  By mixing cocoon and water, the enzyme in the cocoon is extracted into water. The koji used in the method of the present invention may be commercially available or may be cultured as described above. The koji can be selected from the group consisting of rice koji, wheat koji, and soybean koji. The rice bran is preferably rice bran. The water may be any of soft water, intermediate water and hard water. Hard water refers to water having a hardness of 20 ° or more, intermediate water refers to water having a hardness of 10 ° to less than 20 °, and soft water refers to water having a hardness of less than 10 °. The water is preferably soft water or intermediate water, more preferably soft water. The water can also be ion exchange water, ground water, well water, tap water, distilled water, sterilized water, and the like.

  The amount of water mixed with the soot is preferably about 1 or more times the weight of the soot, more preferably about 2 or more times, and more preferably about 2.5 or more times. The amount of water mixed with the soot is preferably no more than about 10 times the weight of the soot, more preferably no more than about 5 times, and even more preferably no more than about 4 times.

  The temperature of the mixture during extraction is preferably about 0 ° C or higher, more preferably about 5 ° C or higher, further preferably about 7 ° C or higher, and most preferably about 10 ° C or higher. The temperature of the mixture can be, for example, about 15 ° C. or higher, about 20 ° C. or higher, and the like. The temperature of the mixture is preferably about 50 ° C. or less, more preferably about 40 ° C. or less, particularly preferably about 30 ° C. or less, and most preferably about 25 ° C. or less. If the temperature of the mixture at the time of extraction is too high, the enzyme in the koji may be deactivated.

  The extraction time is preferably about 10 minutes or more, more preferably about 20 minutes or more, even more preferably about 30 minutes or more, even more preferably about 40 minutes or more, and particularly preferably about 50 minutes. More preferably, it is about 1 hour or more. The extraction time is preferably about 6 hours or less, more preferably about 5 hours or less, even more preferably about 4 hours or less, particularly preferably about 3 hours or less, and most preferably about 2-2. .5 hours. If the extraction time is about 1 hour, the extracted enzyme amount is almost saturated, so that the working efficiency is lowered even if the extraction time is increased. If the extraction time is too long, the soot may absorb water and the yield may decrease. Therefore, it is preferable that the extraction time is moderately short. In this extraction step, the extraction time is preferably adjusted so that the Brix sugar content of the extract obtained by extraction is about 5 to about 7. The Brix sugar content can be measured by a Brix meter. In order to improve extraction efficiency, it is preferable to stir during extraction. Stirring is preferably performed slowly and intermittently so that the wrinkles do not collapse. This extraction step can be performed with stirring in the extraction tank.

  After extraction, the extract is separated from the mixture. Separation can be performed according to methods known in the art. Examples of the separation method include filtration and centrifugation. Filtration with a mesh is preferred. In certain embodiments, it is preferred that the mixture is finally filtered through 60 mesh. In this step, a discharge pump, a separation net, or the like can be used. Filtration may be performed under normal pressure or reduced pressure filtration. The extract obtained at this time is also referred to as “boiled extract”. When rice bran is used as the koji, the obtained koji extract is a rice koji extract. When wheat straw is used as straw, the resulting straw extract is wheat straw extract. When soybean koji is used as koji, the koji extract obtained is a koji koji extract. The koji extract is preferably a rice koji extract. In certain embodiments, the yield of koji extract obtained will be greater than or equal to 65% by weight of the water used.

  The resulting extract and excipient (eg, sugar alcohol or non-cariogenic saccharide) are then mixed to obtain an extract excipient mixture (eg, extract sugar-alcohol mixture or extract non-cariogenic saccharide mixture) Is obtained. Here, the excipient is dissolved in the extract. The temperature of the mixture when dissolving the excipient is preferably about 0 ° C. or higher, more preferably about 5 ° C. or higher, more preferably about 7 ° C. or higher, and most preferably about 10 ° C. or higher. . The temperature of the mixture can be, for example, about 15 ° C. or higher, about 20 ° C. or higher, and the like. The temperature of the mixture is preferably about 50 ° C. or less, more preferably about 40 ° C. or less, particularly preferably about 30 ° C. or less, and most preferably about 25 ° C. or less. When the weight of the koji used for extraction is 100 parts by weight, the amount of the excipient to be mixed is preferably about 10 parts by weight or more, more preferably about 20 parts by weight or more, and still more preferably about It is 30 parts by weight or more, particularly preferably about 40 parts by weight or more, and most preferably about 50 parts by weight or more. When the weight of the koji used for extraction is 100 parts by weight, the amount of excipient mixed is preferably about 100 parts by weight or less, more preferably about 90 parts by weight or less, and still more preferably about 80 parts by weight or less, particularly preferably about 70 parts by weight or less, and most preferably about 60 parts by weight or less.

  The extract excipient mixture is then dried (preferably lyophilized or spray dried) to obtain a dried koji extract. Drying is a drying method in which the activity of the enzyme in the koji extract remains. Such drying methods are known to those skilled in the art. The drying is preferably freeze drying or spray drying, and more preferably spray drying. Spray drying has the advantage of higher productivity and lower cost than freeze drying. The obtained dried koji extract can be applied to a shifter or the like as necessary to obtain a powder with an appropriate particle size. The dried soot extract is preferably, for example, 50 mesh pass.

(3. Dry rice cake extract)
The dried koji extract of the present invention includes a dried product of a mixture containing koji extract and a non-cariogenic excipient, and has protease activity and tannase activity.

  In the present specification, the term “dried koji extract” refers to a dried koji extract. The dried koji extract is preferably powdered. Since the dried koji extract of the present invention is pulverized so that the enzyme activity remains sufficiently, the enzyme activity is high. Further, since the conventional powder is powdered using cariogenic saccharide as an excipient, the content of cariogenic saccharide is high. However, the dry cocoon extract of the present invention has a non-cariogenic excipient (preferably as an excipient). Uses sugar alcohol or non-cariogenic saccharides), so the content of cariogenic saccharides is low. Therefore, the dry koji extract of the present invention has low caries.

  The protease activity of the dried koji extract of the present invention is preferably about 50 U / g or more, more preferably about 100 U / g or more, even more preferably about 200 U / g or more, and particularly preferably about 300 U / g. / G or more, most preferably about 350 U / g or more. The protease activity of the dried koji extract of the present invention is preferably about 5000 U / g or less, more preferably about 3000 U / g or less, even more preferably about 2000 U / g or less, and particularly preferably about 1500 U / g. / G or less, most preferably about 1000 U / g or less.

  The acidic protease activity of the dried koji extract of the present invention is preferably about 700 U / g or more, more preferably about 1400 U / g or more, even more preferably about 2800 U / g or more, particularly preferably about 4200 U / g or more, most preferably about 4900 U / g or more. The acidic protease activity of the dried koji extract of the present invention is preferably about 70000 U / g or less, more preferably about 42000 U / g or less, even more preferably about 28000 U / g or less, particularly preferably about 21000 U / g or less, and most preferably about 14000 U / g or less.

  The tannase activity of the dried koji extract of the present invention is preferably about 50 U / g or more, more preferably about 200 U / g or more, even more preferably about 300 U / g or more, and particularly preferably about 400 U / g. / G or more, most preferably about 500 U / g or more. The tannase activity of the dried koji extract of the present invention is preferably about 50000 U / g or less, more preferably about 20000 U / g or less, still more preferably about 10,000 U / g or less, particularly preferably about 5000 U. / G or less, most preferably about 2000 U / g or less.

  The content of carious saccharides in the dried koji extract of the present invention is about 50% by weight or less, preferably about 40% by weight or less, more preferably about 30% by weight or less, and further preferably about 25% by weight. % Or less, and most preferably about 20% by weight or less.

  The glucose content of the dried koji extract of the present invention is about 30% by weight or less, preferably about 25% by weight or less, more preferably about 20% by weight or less, and further preferably about 15% by weight or less. And most preferably no more than about 10% by weight.

  The starch content of the dried koji extract of the present invention is about 30% by weight or less, preferably about 25% by weight or less, more preferably about 20% by weight or less, and further preferably about 15% by weight or less. And most preferably no more than about 10% by weight.

  In the production of the dried koji extract of the present invention, koji is extracted at a relatively low temperature for a short time with water or alcohol. Therefore, the amount of sugar extracted from the koji into the extract is small. Therefore, the sugar content in the obtained liquid smoke powder is also small. On the other hand, in the field of sake production, as “koji extract”, “as a medium for cultivating yeast etc. with a liquid obtained by adding water to rice koji and saccharifying it at around 55 ° C. It is also known as “soup soup”. However, the conventional koji extract used in the field of sake production is, for example, a saccharide extract that is added with 4 liters of warm water to one liter of koji and saccharifies at around 55 ° C. for a long time of usually about 6 hours or more. Contains a large amount of sugar (eg, glucose, starch). When a dried koji extract is produced using a koji extract obtained by such a conventional production method as a raw material, the sugar content in the dried koji extract becomes too high. If the sugar content in the dried koji extract is too high, it will cause caries in the oral cavity, which is not preferable.

  When mentioning the carious saccharide content of the dried cocoon extract, the “cariogenic saccharide” refers to a saccharide present in the dried cocoon extract and a carious saccharide. The carious saccharides contained in the koji extract are mainly monosaccharides and disaccharides such as glucose. Therefore, in the present specification, the content of carious saccharide refers to the content of carious monosaccharide and disaccharide. The “sugar” in this case does not include sugar alcohol. The amount of glucose in the dried koji extract can be measured by the mutarotase glucose oxidase method. The content of carious saccharides in the dried koji extract can be measured by quantifying reducing sugars. The quantification of reducing sugar can be measured by a known method.

  In the dried cocoon extract, the cocoon extract is included as a dry product of a mixture containing the cocoon extract and the non-cariogenic excipient.

  The amount of water in the dried koji extract is preferably small, but it is practically difficult to make it zero. Therefore, the lower limit of the amount of water in the dried koji extract is, for example, about 0.001 wt% or more, about 0.005 wt% or more, about 0.01 wt% or more, about 0.05 wt% or more, about 0 0.01% or more, about 0.05% or more, about 0.1% or more, and the like. The water content in the dried koji extract is preferably about 5% by weight or less, more preferably about 4% by weight or less, still more preferably about 3% by weight or less, and particularly preferably about 2% by weight or less. And most preferably no more than about 1% by weight.

  The total content of sugar alcohol and non-cariogenic sugar in the dried koji extract is preferably about 40% by weight or more, more preferably about 50% by weight or more, and further preferably about 60% by weight or more. Particularly preferred is about 70% by weight or more, and most preferred is about 80% by weight or more. The total content of sugar alcohol and non-cariogenic saccharide in the dried koji extract is preferably about 99% by weight or less, more preferably about 97% by weight or less, and further preferably about 95% by weight or less. Particularly preferably, it is about 90% by weight or less, and most preferably about 85% by weight or less.

(4. Food containing dried koji extract)
The food of the present invention contains the dried koji extract of the present invention, and has a carious saccharide content of 0.5% by weight or less.

  The food of the present invention contains a dried koji extract. Examples of food products of the present invention include gums, candy and tablets. The food of the present invention is preferably a gum, candy or tablet. Examples of candy include hard candy, soft candy and caramel.

  The food of the present invention is preferably in a form that allows sustained release of the enzyme contained in the dried koji extract. It is preferable that the food of the present invention stays in the oral cavity for about 1 minute or more in a normal ingestion manner. The food of the present invention is more preferably a food that stays for about 3 minutes or more, more preferably about 5 minutes or more, particularly preferably about 7 minutes or more, and most preferably about 10 minutes or more. Such a form of food that can stay for a long time can prolong the time during which the dried koji extract is present in the oral cavity, compared to a form of food that immediately passes through the oral cavity (for example, juice). Has the advantage. When the residence time is too short, it is difficult to obtain a stain removal effect or a stain prevention effect.

  The time that the gum is present in the oral cavity depends on the size and raw material of the gum, but also greatly depends on the eating method of the user. Therefore, it is preferable to chew gum for a sufficient amount of time in the oral cavity (for example, about 1 minute or more, about 3 minutes or more, about 5 minutes or more, about 7 minutes or more, about 10 minutes or more). Is preferably displayed.

  Gum refers to food containing a gum base. Examples of gum bases include chicle, vinyl acetate, ester gum, polyisobutylene and styrene butadiene rubber. Examples of gums include chewing gums such as board gums and bubble gums. The gum of the present invention preferably contains no or even a small amount of carious saccharides. Examples of raw materials used for gums include: gum base, gelatin, flavoring and coloring. The gum may also contain sugar alcohols or non-cariogenic sugars. The sugar alcohol or non-cariogenic saccharide is more preferably selected from indigestible dextrin, palatinose, trehalose, lactitol, reduced palatinose, palatinose, maltitol, erythritol, sorbitol, xylitol and mannitol. The formulation of the gum can be according to formulations known in the art. Gum is suitable as the food of the present invention like a tablet because an active ingredient can be retained in the oral cavity for a long time.

  The time it takes for the candy and tablet to start melting in the oral cavity and finish melting depends on the size and ingredients of the candy and tablet and the eating method of the eater. One skilled in the art can arbitrarily design and manufacture candy and tablets suitable to achieve the desired time from when the candy and tablet begin to melt to when they finish melting.

  In the present specification, “tablet” refers to a tablet material produced by compressing a tablet material into a certain shape. Such a tablet is also called a compressed tablet after its manufacturing method. In the present specification, the tablet may be a tablet for food use or a tablet for pharmaceutical use (that is, a tablet). Examples of tablets for food use include tablets as confectionery called tablet confectionery and tablets as health food (for example, supplements). In the present specification, a tablet having a cavity in the center is called a troche.

  Examples of raw materials used in tablets include: non-cariogenic saccharides such as indigestible dextrin, palatinose, trehalose; sugars such as reduced palatinose, lactitol, maltitol, erythritol, sorbitol, xylitol, mannitol Alcohol; calcium carbonate, calcium phosphate, calcium sulfate, powdered cellulose, emulsifier, flavoring and coloring. The tablet preferably contains an excipient in addition to the dried koji extract. Examples of excipients include the non-cariogenic saccharides and sugar alcohols described above. The excipient preferably comprises a sugar alcohol or a non-cariogenic saccharide, more preferably a sugar alcohol or a non-cariogenic sugar, and even more preferably an indigestible dextrin, palatinose, trehalose, lactitol, reduced palatinose, maltitol, erythritol. , Sorbitol, xylitol and mannitol. These excipients are characterized by excellent taste quality or excellent industrial aptitude.

  In the present specification, the term “candy” refers to a solid food produced by a method including a step of simmering sugar or sugar alcohol and water using sugar or sugar alcohol as a main raw material. Candy is classified into soft candy and hard candy. The hardness of the candy is affected by the candy material and the cooking temperature.

  Examples of soft candy include caramel, nougat and marshmallow.

  Examples of hard candy include drops, butterscotch, toffee, liters (eg, peanut brittle) and bekko candy. Illustrating the case where sugar and starch syrup are mainly used as the sugar, the typical boiling temperature of the drop is about 145 ° C. Illustrating the case where sugar and starch syrup are mainly used as sugar, a typical boiling temperature of butterscotch is about 145 ° C., including about 4% to about 6% by weight of butter and the like. Illustrating the case where sugar and starch syrup are mainly used as sugar, a typical boiling temperature of peanut brittle is about 146 ° C., and includes peanut, baking soda and the like. As an example of the case where sugar and starch syrup are mainly used as the sugar, the typical boiling temperature of the beet koji is about 150 ° C to 160 ° C. A typical sugar candy candy solid content ratio is sugar 40 to 60: candy 40 to 60, and includes dairy products, fats and oils, and the like.

  In the candy of the present invention, it is preferable that a cariogenic saccharide is not contained in the raw material or even if it is contained in a very small amount. Examples of ingredients used in candy include the following: flour, salt, agar, gelatin, peanuts, shortening, butter, acidulants, flavors and colorings. The sugar alcohol or non-cariogenic saccharide is more preferably selected from lactitol, reduced palatinose, maltitol, erythritol, sorbitol, xylitol and mannitol. The candy can be blended according to a blend known in the art. Since candy can retain active ingredients in the oral cavity for a long time, it is suitable as a food of the present invention like a tablet.

  The food of the present invention is preferably in the form of gum, candy, tablet, etc., since it is considered that teeth and saliva and enzymes (for example, protease and tannase) in the dried salmon extract come into contact with each other for a long time. .

  As long as the food of the present invention does not adversely affect the stain-removing action and the stain-preventing action and does not cause caries, the food may contain any ingredient that the food normally contains in addition to the dried koji extract. Examples of such components include excipients such as dietary fiber; sweeteners; fragrances; colorants; acidulants;

  In the food of the present invention, the dried koji extract may be uniformly dispersed or localized. For example, when the food is a candy, the dried koji extract may be used as a candy center or may be uniformly dispersed in the candy.

  The food of the present invention can take any shape. The food of the present invention may be in a homogeneous lump shape or may have a shape having a two-layer or three-layer structure.

  When the food of the present invention is a gum, candy or tablet, it is possible to control the duration of action of the dried koji extract by adjusting its size, dissolution rate and disintegration rate.

  The food of the present invention may also be in the form of sugar-coated tablets. In this case, it is preferable that the uncoated tablet contains an inhibitor of dried koji extract, and the sugar-coated layer contains the dried koji extract. This food can be prepared according to methods known to those skilled in the art of producing sugar-coated tablets. For example, the sugar-coating layer may be performed manually by a method called “octopus” or by a method called dragee.

  In the case of a sugar-coated tablet, the weight of the sugar-coated tablet is preferably about 0.05 g or more, more preferably about 0.1 g or more, further preferably about 0.2 g or more, and most preferably about 0.5 g or more. is there. In the case of a sugar-coated tablet, the weight of the sugar-coated tablet is preferably about 10 g or less, more preferably about 8 g or less, still more preferably about 5 g or less, and most preferably about 3 g or less. The weight of the uncoated tablet in the sugar-coated tablet is preferably about 0.005 g or more, more preferably about 0.01 g or more, and most preferably about 0.1 g or more. The weight of the uncoated tablet in the sugar-coated tablet is preferably about 1.5 g or less, more preferably about 1.0 g or less, and most preferably about 0.5 g or less. The weight of the sugar-coating layer in the sugar-coated tablet is preferably about 0.001 g or more, more preferably about 0.01 g or more, and most preferably about 0.05 g or more. The weight of the sugar-coating layer in the sugar-coated tablet is preferably about 1.0 g or less, more preferably about 0.5 g or less, and most preferably about 0.3 g or less.

  When the food of the present invention is a gum, the weight of the food of the present invention is preferably about 0.05 g or more, more preferably about 0.1 g or more, further preferably about 0.2 g or more, Most preferably, it is about 0.5 g or more. When the food of the present invention is a gum, candy or tablet, the weight of the food of the present invention is preferably about 10 g or less, more preferably about 8 g or less, even more preferably about 5 g or less, most preferably Is about 3 g or less.

  When the food of the present invention is a candy, the weight of the food of the present invention is preferably about 0.05 g or more, more preferably about 0.1 g or more, further preferably about 0.2 g or more, Most preferably, it is about 0.5 g or more. When the food of the present invention is a gum, candy or tablet, the weight of the food of the present invention is preferably about 10 g or less, more preferably about 8 g or less, even more preferably about 5 g or less, most preferably Is about 3 g or less.

  When the food of the present invention is a tablet, the weight of the food of the present invention is preferably about 0.05 g or more, more preferably about 0.1 g or more, still more preferably about 0.2 g or more, Most preferably, it is about 0.5 g or more. When the food of the present invention is a gum, candy or tablet, the weight of the food of the present invention is preferably about 10 g or less, more preferably about 8 g or less, even more preferably about 5 g or less, most preferably Is about 3 g or less.

  When the food of the present invention is a gum, candy or tablet, it may be licked one at a time, or a plurality (eg, 2 to 10) may be licked at a time. When licking a plurality at a time, the plurality may be licked in the mouth at once, or the plurality may be licked one by one.

  The amount of the dried koji extract contained in the food of the present invention can be any amount as long as a stain removal action or a stain prevention action is obtained by ingesting the food. Such an amount can be readily determined by one skilled in the art. The amount of the dried koji extract contained in the food of the present invention is preferably about 0.01% by weight or more, more preferably about 0.05% by weight or more, further preferably about 0.00%, based on the weight of the food. 1 wt% or more, even more preferably about 0.5 wt% or more, even more preferably about 1 wt% or more, even more preferably about 5 wt% or more. The amount of the dried koji extract in the food product may be, for example, about 10% by weight or more, about 15% by weight or more, about 20% by weight or more based on the weight of the food product. There is no particular upper limit on the amount of the dried koji extract in the food, but based on the weight of the food, for example, about 80% by weight or less, about 70% by weight or less, about 60% by weight or less, about 50% by weight or less, about It may be 40% by weight or less, about 30% by weight or less.

  The protease activity of the food of the present invention is preferably about 5 U / g or more, more preferably about 10 U / g or more, even more preferably about 20 U / g or more, and particularly preferably about 30 U / g or more. And most preferably about 50 U / g or more. The protease activity of the food of the present invention is preferably about 500 U / g or less, more preferably about 400 U / g or less, even more preferably about 300 U / g or less, particularly preferably about 200 U / g or less. And most preferably about 100 U / g or less.

  The acid protease activity of the food product of the present invention is preferably about 70 U / g or more, more preferably about 140 U / g or more, even more preferably about 280 U / g or more, particularly preferably about 420 U / g. More preferably, it is about 700 U / g or more. The acid protease activity of the food product of the present invention is preferably about 7000 U / g or less, more preferably about 5600 U / g or less, even more preferably about 4200 U / g or less, particularly preferably about 2800 U / g. Or less, most preferably about 1400 U / g or less.

  The tannase activity of the food of the present invention is preferably about 10 U / g or more, more preferably about 40 U / g or more, even more preferably about 60 U / g or more, and particularly preferably about 80 U / g or more. And most preferably about 100 U / g or more. The tannase activity of the food of the present invention is preferably about 10,000 U / g or less, more preferably about 4000 U / g or less, still more preferably about 2000 U / g or less, and particularly preferably about 1000 U / g or less. And most preferably about 400 U / g or less.

  The content of carious saccharide in the food of the present invention is about 0.5% by weight or less, preferably about 0.3% by weight or less, more preferably about 0.2% by weight or less, and still more preferably Is about 0.1 wt% or less, most preferably about 0.05 wt% or less.

  The food of the present invention can be used for both humans and non-human animals (eg, dogs, cats, mice, rats, rabbits, monkeys, horses, cows, etc.). The food of the present invention is preferably for human use. When the food of the present invention is used for animals, the animals are preferably pets, more preferably dogs or cats, and most preferably dogs. When the food of the present invention is for animals, the food of the present invention is a pet chewing gum. Pet chewing gum compositions and methods of manufacture are known to those skilled in the art of pet chewing gum.

  The food of the present invention can be prepared according to methods known in the art. The food of the present invention is preferably produced by a method that does not substantially impair the stain removing action or the stain preventing action of the dried koji extract. For example, when the dried candy extract is contained in the candy, the candy material is boiled according to a normal method, and then the boiled material is cooled, preferably about 60 ° C. or less, more preferably about 50 ° C. or less, Even more preferably, when the temperature reaches about 40 ° C. or lower, the dried koji extract is added to the candy material. For example, the candy of this invention can be manufactured according to the method as described in an international publication 2007/105661 pamphlet.

  The food of the present invention may contain other components useful for the stain removing action or the stain preventing action as necessary as long as they do not adversely affect the stain removing action, the stain preventing action and the caries prevention.

  The gum, candy or tablet of the present invention may also contain additives usually contained in the gum, candy and tablet, as necessary, as long as they do not adversely affect the stain removal action, stain prevention action and caries prevention. Examples of the additive include a binder, a pH adjuster, a fragrance, a sweetener, a colorant, an excipient, a disintegrant, a lubricant, a fluidizing agent, and a coating agent. These additives are well known in the art. If the gum, candy or tablet is for food use, such additives are described, for example, in the Food Additives Official Document. Such additives are described, for example, in the Japanese Pharmacopoeia when the gum, candy or tablet is for pharmaceutical use.

  The term “binder” as used herein refers to an additive used to provide a binding force to a mixture of ingredient powders to produce a stable tablet or granule. The tablet of the present invention does not necessarily contain a binder, but can be added when the tablet has insufficient hardness. The binder is preferably cellulose, gum arabic, gelatin or the like. The binder improves the hardness of the tablet by bonding between substances contained in the tablet. The hardness of a tablet obtained by tableting using only a specific substance is an indicator of the property of the substance as a binder. The higher the hardness, the stronger the bond strength.

(5. Method for producing food containing dried koji extract)
The food of the present invention can be produced using the techniques and equipment used in conventional food production as they are. For example, when the food is a tablet, the tablet material (for example, water-soluble saccharide and other ingredients) is mixed and directly compressed, or the tablet material is granulated wet or dry by mixing separately or mixed. A method of locking can be adopted.

  In one embodiment, the tablet of the present invention is preferably manufactured by mixing powdered tablet materials and directly tableting.

  In another embodiment, the tablet of the present invention is preferably manufactured by tableting after mixing the granular tablet material.

  The particle size of the dried koji extract is arbitrary, but is preferably about 50 μm or more in average diameter, more preferably about 75 μm or more, and most preferably about 90 μm or more. The particle size of the dried soot extract is preferably about 500 μm or less in average diameter, more preferably about 350 μm or less, and most preferably about 250 μm or less. The average diameter is a weight average diameter.

(6. Eating method of food containing dried koji extract)
The intake amount, intake frequency, and intake period of the food of the present invention are determined according to the condition of the intake person, the preference of the intake person, and the like.

  The intake of the gum of the present invention is preferably about 0.1 g or more, more preferably about 0.2 g or more, still more preferably about 0.5 g or more, and still more preferably about 0.1 g or more. 1 g or more. There is no particular upper limit on the amount of the gum of the present invention, but for example, about 1000 g or less, about 750 g or less, about 500 g or less, about 250 g or less, about 100 g or less, about 50 g or less, about 40 g or less, about 30 g per serving. Hereinafter, about 20 g or less, about 10 g or less, about 7.5 g or less, about 5 g or less, about 4 g or less, about 3 g or less, about 2 g or less, about 1 g or less.

  The intake of the candy of the present invention is preferably about 0.1 g or more, more preferably about 0.2 g or more, still more preferably about 0.5 g or more, and still more preferably about 0.1 g or more. 1 g or more. There is no particular upper limit on the intake of the candy of the present invention, but for example, about 1000 g or less, about 750 g or less, about 500 g or less, about 250 g or less, about 100 g or less, about 50 g or less, about 40 g or less, about 30 g per serving. Hereinafter, about 20 g or less, about 10 g or less, about 7.5 g or less, about 5 g or less, about 4 g or less, about 3 g or less, about 2 g or less, about 1 g or less.

  The intake of the tablet of the present invention is preferably about 0.1 g or more, more preferably about 0.2 g or more, still more preferably about 0.5 g or more, and still more preferably about 0.1 g or more. 1 g or more. There is no particular upper limit on the intake of the tablet of the present invention, but for example, about 1000 g or less, about 750 g or less, about 500 g or less, about 250 g or less, about 100 g or less, about 50 g or less, about 40 g or less, about 30 g per time. Hereinafter, about 20 g or less, about 10 g or less, about 7.5 g or less, about 5 g or less, about 4 g or less, about 3 g or less, about 2 g or less, about 1 g or less.

  The intake frequency of the tablet of the present invention can be arbitrarily set. For example, at least once a week, at least twice a week, at least 3 times a week, at least 4 times a week, at least 5 times a week, at least 6 times a week, at least 7 times a week It may be once a day or more, twice a day or more, three times a day or more. There is no upper limit on the intake frequency of the tablet of the present invention, for example, 3 times or less per day, 2 times or less per day, 1 time or less per day, 7 times or less per week, 6 times or less per week, 5 times per week Or less, 4 or less per week, 3 or less per week, 2 or less per week, 1 or less per week, or the like.

  The timing of intake of the food of the present invention may be before a meal, after a meal, or between meals, but is preferably after a meal. Pre-meal means from about immediately before meal to about 30 minutes before eating, post-meal means from immediately after meal to about 30 minutes after meal, and between meals is about 2 after eating. It means the time about two hours or more before the next meal after more than an hour.

  The intake period of the food of the present invention can be arbitrarily determined. The food of the present invention can be ingested preferably for about 1 day or more, more preferably for about 3 days or more, and most preferably for about 5 days or more. The food of the present invention can be ingested preferably about 1 month or less, more preferably about 2 weeks or less, and most preferably about 10 days or less. If necessary, the food of the present invention may be consumed almost permanently.

  The food of the present invention is preferably retained in the oral cavity without swallowing during ingestion. The time for allowing the food of the present invention to stay in the oral cavity is preferably about 10 seconds or longer, more preferably about 1 minute or longer, and further preferably about 3 minutes or longer. The time for retaining the tablet of the present invention in the oral cavity is preferably about 30 minutes or less, more preferably about 20 minutes or less, and still more preferably about 10 minutes or less. When the residence time is too short, it is difficult to obtain a stain removal effect or a stain prevention effect. The food of the present invention is preferably licked to the end without chewing.

(7. Composition for oral cavity containing dried koji extract)
The composition for oral cavity of the present invention contains the dry cocoon extract of the present invention, and the carious saccharide content is 0.5% by weight or less.

  The oral composition of the present invention is preferably a dentifrice or a mouthwash.

  The oral composition of the present invention is preferably a troche or gel.

  When the oral composition is used in a form such as a dentifrice, mouthwash, gel, etc. that acts at the same concentration without being diluted in the oral cavity, the oral composition of the present invention The amount of the dried koji extract is preferably about 0.01% by weight or more, more preferably about 0.05% by weight or more, and further preferably about 0.1% by weight, based on the weight of the oral composition. Or more, more preferably about 0.5% by weight or more, even more preferably about 1% by weight or more, even more preferably about 5% by weight or more; Based on the weight of the food, it may be, for example, about 10% by weight or more, about 15% by weight or more, about 20% by weight or more. There is no particular upper limit on the amount of the dried koji extract in the oral composition of the present invention when used in a form that acts as it is, but for example, about 80% by weight based on the weight of the food. Hereinafter, it may be about 70% by weight or less, about 60% by weight or less, about 50% by weight or less, about 40% by weight or less, about 30% by weight or less. When the composition for oral cavity is a composition intended to be used diluted in the oral cavity, the ingredients are blended in consideration of the dilution ratio. For example, in the case of an oral composition intended to be diluted about 20 times, it is blended at a concentration of 20 times.

  When the oral composition of the present invention is a troche, the weight of the oral composition of the present invention is preferably about 0.05 g or more, more preferably about 0.1 g or more, and still more preferably about 0.2 g or more, and most preferably about 0.5 g or more. When the oral composition of the present invention is a troche, the weight of the oral composition of the present invention is preferably about 10 g or less, more preferably about 8 g or less, and even more preferably about 5 g or less. Most preferably, it is about 3 g or less.

  When the composition for oral cavity of the present invention is a troche, one tablet may be licked at a time, or a plurality (for example, 2 to 10) may be licked at a time. When licking a plurality at a time, the plurality may be licked in the mouth at once, or the plurality may be licked one by one.

  The protease activity of the oral composition of the present invention is preferably about 2.5 U / g or more, more preferably about 5 U / g or more, still more preferably about 10 U / g or more, particularly preferably. About 15 U / g or more, most preferably about 25 U / g or more. The protease activity of the oral composition of the present invention is preferably about 250 U / g or less, more preferably about 200 U / g or less, even more preferably about 150 U / g or less, and particularly preferably about 100 U / g. / G or less, most preferably about 50 U / g or less.

  The acidic protease activity of the oral composition of the present invention is preferably about 35 U / g or more, more preferably about 70 U / g or more, even more preferably about 140 U / g or more, and particularly preferably about 210 U / g or more, most preferably about 350 U / g or more. The acidic protease activity of the oral composition of the present invention is preferably about 3500 U / g or less, more preferably about 2800 U / g or less, still more preferably about 2100 U / g or less, particularly preferably about 1400 U / g or less, most preferably about 700 U / g or less.

  The tannase activity of the oral composition of the present invention is preferably about 5 U / g or more, more preferably about 20 U / g or more, still more preferably about 30 U / g or more, particularly preferably about 40 U. / G or more, most preferably about 50 U / g or more. The tannase activity of the food of the present invention is preferably about 5000 U / g or less, more preferably about 2000 U / g or less, even more preferably about 1000 U / g or less, and particularly preferably about 500 U / g or less. And most preferably about 200 U / g or less.

  The content of carious saccharide in the oral composition of the present invention is about 0.5% by weight or less, preferably about 0.3% by weight or less, more preferably about 0.2% by weight or less. More preferably, it is about 0.1% by weight or less, and most preferably about 0.05% by weight or less.

  The oral composition of the present invention can be used for humans and non-human animals (eg, dogs, cats, mice, rats, rabbits, monkeys, horses, cows, etc.). When the oral composition of the present invention is used for animals, the animal is preferably a pet, more preferably a dog or cat, and most preferably a dog.

(8. Method for producing oral composition containing dried salmon extract)
The oral composition of the present invention can be produced according to a method known in the art. The composition for oral cavity of the present invention is preferably produced by a method which does not substantially impair the stain removing action or the stain preventing action of the dried koji extract. For example, when a dry wrinkle extract is contained in a dentifrice, mouthwash, gel, or troche, it can be produced by mixing the dry wrinkle extract with the respective materials of a dentifrice, a mouthwash, and a gel. .

(9. Method of using composition for oral cavity)
In certain embodiments, the oral compositions of the present invention can be used as follows. First, the composition for oral cavity of the present invention is applied to a desired tooth surface (for example, an initial caries portion or a healthy portion). This composition is preferably applied to the tooth surface using a device such as a roller or a brush. During and after application of this composition, it may be in contact with saliva, and measures may be taken to reduce contact with saliva so that the enzyme in the applied dry koji extract will not escape as much as possible. Good. The time taken to reduce the contact with saliva is preferably continued for about 5 minutes or more, more preferably about 10 minutes or more, most preferably about 15 minutes or more from the start of application of these compositions. preferable. There is no particular upper limit to the time for taking measures to reduce contact with saliva, for example, about 1 hour or less, about 45 minutes or less, about 30 minutes or less, about 25 minutes or less after starting to apply these compositions, Such as about 20 minutes or less. By taking measures to reduce contact with saliva, the stain removal action and / or the stain prevention action can be significantly promoted.

  The oral composition of the present invention is preferably retained in the oral cavity for a certain period of time when administered into the oral cavity. The time for retaining the oral composition of the present invention in the oral cavity is preferably about 1 minute or longer, more preferably about 2 minutes or longer. More preferably, it is about 3 minutes or more, and particularly preferably about 5 minutes or more. In one preferred embodiment it is about 10 minutes or more, and in a more preferred embodiment it is about 15 minutes or more. There is no particular upper limit to the time for the composition of the present invention to stay in the oral cavity, and it may be, for example, about 1 hour or less, about 50 minutes or less, about 40 minutes or less, about 30 minutes or less, about 20 minutes or less. If the residence time is too short, it is difficult to obtain a stain removal effect and a stain prevention effect.

  Examples of oral compositions other than food include dentifrices, mouthwashes (also referred to as mouthwashes), troches, gels, sprays, pastes, ointments and the like. Examples include tablets, pills, powders, solutions, suspensions, emulsions, granules, capsules and the like. It is also possible to use a form such as a wiping cloth in which a nonwoven fabric or the like is impregnated with these liquid agents, or a form such as a cotton swab.

  The oral composition of the present invention is usually sold in a container or packaged. This container may be a commonly used container such as plastic. This packaging may be a commonly used packaging such as paper, plastic, cellophane and the like. In this container or package, instructions on the intake amount, intake timing, intake method of the oral composition of the present invention (for example, in the case of gum, “it is preferable to continue chewing two tablets for about 20 minutes or more”), etc. Preferably it is described. Alternatively, an instruction sheet in which such an instruction is described may be inserted.

(10. Measuring method)
Various enzyme activities can be measured according to the following methods. Methods such as pellicle decomposition effect and pellicle deposition suppression effect are also described below.

  The activity of the enzyme contained in the dried koji extract, food and oral composition of the present invention can be measured by methods known in the art. For example, when the food and oral composition of the present invention are solids such as candy, the food and oral composition of the present invention are pulverized and dissolved in an appropriate buffer solution for enzyme activity measurement. Enzyme activity is measured. Care should be taken not to inactivate the enzyme by applying excessive heat when crushing the food and oral composition of the present invention. The temperature of the buffer solution during dissolution of the food and oral cavity composition of the present invention is preferably about 4 ° C. Methods for measuring the activity of various enzymes are known. If the same method is used for the enzyme before being added to the food and oral composition of the present invention and the enzyme in the food and oral composition of the present invention, any appropriate method can be used.

(10.1 Method for measuring protease activity)
The protease activity in the dried koji extract, food and oral composition of the present invention is preferably the activity of endoprotease measured according to the manufacturer's recommended method using PROTAZYME AK TABLETS from Megazyme. In this measurement method, the activities of all endoproteases that are all active against casein can be measured. Examples of such proteases are papain, proteinase K, trypsin, bovine pancreatic protease, subtilisin A, Bacillus subtilis protease and Aspergillus niger protease.

(10.2 Tannase activity measurement method)
The activity of tannase is described in the reference; http: // www. kikkoman. co. jp / bio / j / rinsyou / pdf / 65_tah. pdf and http: // www. kikkoman. co. jp / bio / j / skaku / items / skako01. It can be measured with reference to html.

  Regarding the activity of tannase, the activity of hydrolyzing an ester bond in 1 μmol of tannic acid per minute in an environment of 30 ° C. and pH 5.5 is defined as 1 unit (1 U).

(10.2.1 Reagent Preparation)
Prepare the reagents as follows:
Reagent A: citrate buffer Add 800 ml of distilled water to 10.5 g of citric acid and dissolve. Adjust to pH 5.5 with 1N NaOH and fill up to 1000 ml.
Reagent B: substrate solution (tannic acid solution)
Dissolve 175 mg of tannic acid in 50 ml of Reagent A. It does not dissolve completely, but becomes a suspension. Shake well before use without filtering.
Reagent C: aqueous ethanol solution Add 95 ml of distilled water to 900 ml of ethanol.
Sample: Enzyme solution Dissolve the sample in ice-cooled Reagent A immediately before use. For accurate measurement, it is preferable to adjust to 2.6 to 2.9 U / ml.

(10.2.2 Measurement)
(10.2.2.2.1) Prepare a 15 ml tube or a test tube with a lid;
(10.2.2.2) Reagent B 1.0 ml is placed in a tube;
(10.2.2.2.3) Cap the tube and incubate at 30 ° C. for 5 minutes;
(10.2.2.4) Add 250 μl of sample to the tube and stir well. Reagent A is added to the blank instead of the sample;
(10.2.5.) Cap the tube and incubate at 30 ° C. for 15 minutes;
(10.2.2.2.6) Add Reagent C 5.0 ml to the tube and stir;
(10.2.2.2.7) Place 250 μl of each solution in separate tubes;
(10.2.2.2.8) Add Reagent C 5.0 ml to the tube;
(10.2.2.9) Stir well and measure absorption at 310 nm.

(10.2.3 Calculation of enzyme activity)
Volume activity (U / ml)
= [(A ₀ −A _s ) × 20.3 × 1.0 × 1.04 × df] / (0.71 × 0.25 ml × 15 min)
A ₀ ; Absorbance of blank As _s ; Absorbance of sample df; Dilution factor
20.3; amount of tannic acid per 1.0 ml of Reagent B (μmol)
0.71; change in absorbance when hydrolysis of 20.3 μmol of tannic acid was completed under this assay condition 1.04; optimization factor according to the method of Iibuchi et al.

(10.3 Method for measuring enzyme activity: National Tax Agency prescribed analysis method)
(10.3.1) Acidic protease (10.3.1.1) Acidic protease 1-1. Reagent (1) McKilvain buffer (pH 3.0)
Solution A: M / 5 Disodium Phosphate Solution Dissolve 71.63 g of disodium phosphate (Na ₂ HPO ₄ · 12H ₂ O) in water to make 1 liter.
Solution B: the M / 10 citric acid solution of citric acid _{(C 6 H 8 O 7 ·} H 2 O) 21g to 1 liter dissolved in water.

  Mixing at a ratio of 4 ml of liquid A and 16 ml of liquid B, the pH becomes almost predetermined. If the pH does not reach 3.0 correctly, use solution A or solution B.

(2) Casein solution Take 2 g of casein, add 5 ml of 10-fold diluted lactic acid, add about 50 ml of water, and stir while heating on a wire mesh until completely dissolved in a cloudy state.

  Once boiled and cooled, 20 ml of pH 3.0 McKilvein buffer is added, and water is added to make the whole volume 100 ml.

(3) 2M / 5 trichloracetic acid solution (TCA solution)
Dissolve 65.4 g of trichloracetic acid in water to make 1 liter.

(4) Phenol reagent (Forin / Thiocult-Reagent)
Use a commercial product diluted 5 times.

(5) 2M / 5 sodium carbonate solution 42.4 g of sodium carbonate is dissolved in water to make 1 liter.

(6) Tyrosine standard solution Take 10.0 mg of L-tyrosine (special grade), add 1 ml of 1N hydrochloric acid to make a total volume of 100 ml, and use this as the stock solution.

  In the measurement, water is added to the stock solution to make a standard solution series of 20-100 μg / ml of tyrosine.

(7) Preparation of calibration curve Add 5 ml of sodium carbonate solution and 1 ml of phenol reagent to each of the above-mentioned test solutions, and perform color development at 40 ° C. for 30 minutes. Using a solution containing no tyrosine as a control, the absorbance at 660 nm is measured in a 10 mm-thick cell to prepare a calibration curve.

1-2. Preparation of enzyme solution 50 ml of sodium chloride solution is added to 10 g of solid koji, and leached while being shaken occasionally in a low temperature room (5 ° C. or lower) overnight or at room temperature (15 to 20 ° C.) for 3 hours. 10 ml of the filtrate is put into a dialysis membrane, dialyzed overnight against M / 100 acetate buffer at a low temperature, and then made up to 20 ml with water to make an enzyme solution.

1-3. Test procedure 1.0 ml of pH 3.0 McKilvein buffer is added to 1.5 ml of casein solution and preheated to 40 ° C. To this, 0.5 ml of enzyme solution is added and reacted at 40 ° C. for 60 minutes. Then, 3 ml of TCA solution is added to stop the reaction, and the precipitate is filtered off.

  5 ml of sodium carbonate solution and 1 ml of phenol reagent are added to 1 ml of the filtrate, color is developed at 40 ° C. for 30 minutes, and the absorbance at 660 nm is measured.

  Separately, an enzyme solution is added as a control immediately before the addition of the TCA solution, and the absorbance is measured by the same operation as described above.

  The absorbance difference E between the test solution and the control solution is determined. However, when E becomes 0.3 or more, the enzyme force and E deviate from the linear relationship, so the enzyme solution must be diluted so that E becomes 0.3 or less.

  From the obtained E, the amount of tyrosine produced (y μg) is determined by a calibration curve.

1-4. Indication of activity Acidic protease activity is defined as 1 unit of activity showing a color equivalent to 1 μg of tyrosine for 60 minutes at 40 ° C.

  The acid protease activity of 1 g of koji is determined by the following formula:

  (Note) The enzyme solution may not be dialyzed. In that case, the extraction rate is calculated as 50/10.

(10.3.2) Glucoamylase 3-1. Reagent (1) M / 5 acetate buffer (pH 5.0)
Liquid A: M / 5 acetic acid 11.55 ml of glacial acetic acid is made up to 1 liter with water.
Liquid B: M / 5 sodium acetate solution 27.21 g of sodium acetate (CH ₃ COONa · 3H ₂ O) is dissolved in water to make 1 liter.

  Liquid A and liquid B are prepared and mixed at a ratio of liquid A (5.9 ml) and liquid B (14.1 ml). If the pH does not reach 5.0 correctly, use solution A or solution B.

(2) Sodium chloride solution Dissolve 5 g of sodium chloride in water, add 50 ml of the above buffer solution to make 1 liter with water.
(Note) In the case of black koji-type solid koji (hereinafter referred to as “shochu”), 500 ml of buffer solution is added.

(3) M / 100 acetate buffer (pH 5.0)
After diluting 50 ml of M / 5 acetate buffer (pH 5.0) with water to 1 liter, adjust to pH 5.0.

(4) Starch solution Take 2 g of soluble starch, add an appropriate amount of hot water, stir well and boil for 1-2 minutes, then cool and add water to 100 ml.

(5) 1N sodium hydroxide solution 4 g of sodium hydroxide is dissolved in water to make 100 ml.

(6) 1N hydrochloric acid solution Add water to 8.8 ml of concentrated hydrochloric acid to make 100 ml.

  The concentration is adjusted so that neutralization is possible when equal amounts of 1N sodium hydroxide solution and 1N hydrochloric acid solution are mixed.

3-2. Preparation of enzyme solution 50 ml of sodium chloride solution is added to 10 g of solid koji, and leached while being shaken occasionally in a low temperature room (5 ° C. or lower) overnight or at room temperature (15 to 20 ° C.) for 3 hours. 10 ml of the filtrate is put into a dialysis membrane, dialyzed overnight against M / 100 acetate buffer at a low temperature, and then made up to 20 ml with water to make an enzyme solution.

3-3. Test procedure (1) Enzyme reaction Add 0.2 ml of M / 5 acetate buffer to 1 ml of starch solution and preheat at 40 ° C. for 5 minutes. To this, 0.1 ml of enzyme solution is added and reacted at 40 ° C. for 20 minutes, and 0.1 ml of 1N sodium hydroxide solution is added to stop the reaction. The mixture is then allowed to stand for 30 minutes and neutralized by adding 0.1 ml of 1N hydrochloric acid solution.

  Separately, 0.2 ml of M / 5 acetate buffer was added to 1 ml of starch solution, preheated at 40 ° C. for 5 minutes, 0.1 ml of 1N sodium hydroxide solution was added, and then 0.1 ml of enzyme solution was added. Operate as above.

(2) Determination of glucose The amount of glucose (μg) produced in the reaction solution is measured by 3-4.

3-4 Glucose 3-4-1 Reagent M / 10 Tris-phosphate buffer (pH 7.3):
12.11 g of trishydroxymethylaminomethane is taken, dissolved in about 900 ml of water, adjusted to pH 7.3 with phosphoric acid and made up to 1 l with water.
Dye solution:
Take 81 mg of 4-aminoantipyrine, 500 mg of phenol, and 100 mg of Triton X-100 [polyoxyethylene (10) octylphenyl ether], dissolve in M / 10 Tris-phosphate buffer (pH 7.3) to make 200 ml. .
Enzyme solution:
Take 800 units of glucose oxidase and 2000 units of peroxidase and dissolve in M / 10 Tris-phosphate buffer (pH 7.3) to make 200 ml.
Glucose quantification reagent:
Mix equal amounts of dye solution and enzyme solution. After preparation, if stored in a refrigerator, it is stable for about 3 weeks.
Glucose standard solution:
0.5 g of anhydrous crystalline glucose (special grade) is purified and dissolved with 1 g of benzoic acid to make 1 l. This solution contains 500 μg of glucose in 1 ml.

3-4-2 Test procedure Take 3 ml of glucose determination reagent in a test tube, add and mix 0.1 ml of a sample containing 10 to 100 μg of glucose, and then react at 40 ° C. for 40 minutes. After completion of the reaction, the mixture is allowed to cool to room temperature, and the absorbance (E ₁ ) at 505 nm is measured within 120 minutes.

Separately, a control test using water instead of the sample is performed, and the absorbance (E ₂ ) of the control is measured. Further, the same operation is performed using a glucose standard solution to measure the absorbance (E _S ), and glucose in the sample is obtained by the following formula.

(note)
1: One unit of glucose oxidase is a titer that oxidizes 1 μM β-type glucose per minute under conditions of pH 5.1 and 35 ° C.
2: One unit of peroxidase is a titer that produces 1 mg of purbrogallin from pyrogallol in 20 seconds under the conditions of pH 6.0 and 20 ° C.

3-5. Indication of activity The glucoamylase activity is defined as 1 unit of activity for producing 1 mg of glucose from soluble starch at 40 ° C. for 60 minutes. The glucoamylase activity of 1 g of koji is obtained by the following formula.

(note)
1: In order to eliminate the influence of α-amylase, the degradation rate of starch is 10% or less (produced glucose is about 2 mg or less).
2: An enzyme solution having an α-amylase activity of 200 units / ml or less is used.

(10.3.3) α-Amylase 3-1. Reagent (1) M / 5 acetate buffer (pH 5.0)
Liquid A: M / 5 acetic acid 11.55 ml of glacial acetic acid is made up to 1 liter with water.
Liquid B: M / 5 sodium acetate solution 27.21 g of sodium acetate (CH ₃ COONa · 3H ₂ O) is dissolved in water to make 1 liter.

  Liquid A and liquid B are prepared and mixed at a ratio of liquid A (5.9 ml) and liquid B (14.1 ml). If the pH does not reach 5.0 correctly, use solution A or solution B.

(2) Sodium chloride solution Dissolve 5 g of sodium chloride in water, add 50 ml of the above buffer solution to make 1 liter with water.

(3) Iodine solution 0.1 g of potassium iodide and 50 ml of 10% hydrochloric acid are added to 0.0317 g of iodine and dissolved in water to make 1 l.

(4) Starch solution Take 1 g of soluble starch, dissolve it according to 211-4-1, add 20 ml of the above buffer to adjust to pH 5.0, and add water to make 100 ml.

3-2. Preparation of enzyme solution 50 ml of sodium chloride solution is added to 10 g of solid koji, and leached while being shaken occasionally in a low temperature room (5 ° C. or lower) overnight or at room temperature (15 to 20 ° C.) for 3 hours. 10 ml of the filtrate is put into a dialysis membrane, dialyzed overnight against M / 100 acetate buffer at a low temperature, and then made up to 20 ml with water to make an enzyme solution.

3-3. Test procedure Take 2 ml of starch solution in a test tube and preheat at 40 ° C for 5 minutes. Add 0.1 ml of the enzyme solution to start the reaction, and pipette 0.1 ml of the reaction solution over time (0.5 minutes or every other minute) into a test tube containing 10 ml of iodine solution in advance. , Mix well, colorize at 670 nm through a 10 mm thick cell, keeping the resulting color at 25 ° C., and measure the transmittance T%.

  As a control solution for colorimetric use, 0.1 ml is taken out of 2 ml of water mixed with 0.1 ml of enzyme solution and added to 10 ml of iodine solution.

  A reaction time corresponding to 66% is found out of the series of T% values, and this is set as t minutes.

3-4. Indication of activity The α-amylase activity is expressed as the amount of 1% soluble starch (ml) which is degraded in 30 minutes at 40 ° C. The α-amylase activity of 1 g of koji is determined by the following formula.

(note)
1: The soluble starch used for this was prepared by preparing a starch solution according to 211-5-1, taking 0.1 ml from 2 ml mixed with 0.1 ml water, added to 10 ml iodine solution, and the resulting color. Must be colorimetric through a 10 mm thick cell at 670 nm and the transmission T% should be 18-20.

For starches that do not meet this condition, use the following conversion formula: When the reaction time is t minutes, the iodine coloration before the reaction is T ₀ %, and the iodine coloration after the reaction is T _t %

2: If the T% after the enzyme reaction for 5 minutes is 80 or more, dilute the enzyme solution appropriately.

(10.4 Evaluation of pellicle decomposition effect)
(Test with 10.4.1 Hydroxyapatite (Hap) Powder)
1) Saliva is collected by using a saliva collection tube (SARESTED Salivet series, Sarikids series, Sarisoft series, etc. Here, Sarivet 51.1534 is used).

  2) Centrifuge at 2500 rpm for 4 minutes to unite the saliva collected in the tube once.

  3) Weigh 10 mg of Hap powder into a 2 ml tube.

  4) Add 1 ml of saliva to a tube containing Hap powder and vortex (fully agitate). Add 1 ml of saline to the negative control and vortex in the same way.

  5) Warm the shaking incubator to 37 ° C and incubate overnight (16 hours) with 4 tubes in strong shaking.

  6) Remove from the incubator and centrifuge at 13,000 rpm for 5 minutes to remove the supernatant (saliva).

  7) Add 200 μl of physiological saline, and tap to disperse the powder in the physiological saline.

  8) Centrifuge at 13,000 rpm for 5 minutes to remove the supernatant.

  9) Repeat 7) and 8) (= cleaning step).

  10) Add a solution of the substance whose effect is to be confirmed, such as an enzyme solution, to the tube, and incubate with strong shaking for 20 minutes in a 37 ° C shaking incubator. Saline is added to the positive and negative controls. * A dry rice bran extract A aqueous solution and an aqueous solution of purified enzyme derived from the genus Aspergillus (eg, Amano Enzyme “Protease M Amano SD”) were used at a protein digestion power of 100 U / ml.

  11) Remove from the incubator and centrifuge at 13,000 rpm for 5 minutes to remove the supernatant (enzyme solution).

  12) Add 200 μl of physiological saline, and tap to disperse the powder in the physiological saline.

  13) Centrifuge at 13,000 rpm for 5 minutes to remove the supernatant (washing).

  14) Add 100 μl of physiological saline, and tap to disperse the powder in the physiological saline.

  15) After sonication (15 min), centrifuge at 13,000 rpm for 5 min to quantify protein in the supernatant.

  16) (Protein quantification) A commercially available reagent Bio-Rad Protein assay (Bio-Rad) is used. γ-globulin (2.0 mg / ml) is diluted with an equal volume mixture of physiological saline and 0.1 N NaOH, and adjusted to 0, 25, 50, 100, 150, 200 μg / ml.

  17) Put 10 μl each of the supernatant of 11) and the γ-globulin diluted solution of 12) into a 96-well plate, add 200 μl of Protein assay reagent diluted 5-fold with distilled water, and add 5 μm or more (60 minutes) Within)

  18) Measure the absorbance at 595 nm with a plate reader, create a calibration curve with γ-globulin, and 15) calculate the protein concentration in the supernatant.

  19) (Evaluation criteria) Multiply the volume of the supernatant by the protein concentration of 18) to obtain the amount of protein attached to the Hap powder. At this time, about 40 (μg / 10 mg Hap powder) protein adhesion is seen in the Hap powder (positive control) which is not subjected to the saliva treatment alone and the enzyme treatment.

The higher the amount of protein detected, the lower the pellicle removal effect, and the lower the effect, the higher the effect, and the results were encoded as follows:
×: No pellicle degradation effect (protein amount comparable to positive control)
Δ: Pellicle decomposition effect weak ○: Pellicle decomposition effect 効果: Pellicle decomposition effect strong (protein amount comparable to negative control).

(Test with 10.4.2 Hap disc)
1) Saliva is collected by using a saliva collection tube (SARESTED Salivet series, Sarikids series, Sarisoft series, etc. Here, Sarivet 51.1534 is used).

  2) Centrifuge at 2500 rpm for 4 minutes to unite the saliva collected in the tube once.

  3) Place the Hap discs one by one in a 2 ml tube.

  4) Add 1 ml of saliva to the tube containing the Hap disk, and add physiological saline to the negative control.

  5) Warm the shaking incubator to 37 ° C and incubate overnight (16 hours) with 4 tubes in strong shaking.

  6) Remove from the incubator and remove saliva (or saline).

  7) Put 200 μl of physiological saline into the tube and rinse lightly to remove the physiological saline. Repeat the same operation once more.

  8) Add a solution of an enzyme solution or the like whose effect is to be confirmed to the tube, and incubate with strong shaking for 20 minutes in a 37 ° C shaking incubator. Saline is added to the positive and negative controls. * A dry rice bran extract A aqueous solution and an aqueous solution of purified enzyme derived from the genus Aspergillus (eg, Amano Enzyme “Protease M Amano SD”) were both provided at a protein digestion power of 100 U / ml.

  9) Remove from the incubator and remove the enzyme solution.

  10) Add 200 μl of normal saline, lightly rinse and remove.

  11) The protein on the surface of the Hap disk is stained with CBB (Coomassie Brilliant Blue R-250 Staining Solution, manufactured by Bio-Rad).

  12) (Staining) Add 100 μl of CBB solution to the tube containing the Hap disk and leave it for 2 hours or more.

  13) Rinse excess CBB solution with saline.

  14) Put about 1 ml of decoloring solution (40% MeOH, 10% acetic acid) into a tube and lightly rinse. After about 5 minutes, remove the decolorizing solution, add a new decolorizing solution, and perform the same operation.

  15) Remove the disc from the tube and take a picture with a digital camera. At that time, the image correction color chart CasMatch (Bare Medic Co., Ltd.) is reflected.

  16) (Image processing) Correction is performed using cast matching in the photograph, and then the Lab value is confirmed (using Adobe Photoshop). The larger the L value (brightness), the brighter. a value (red to green index). b value (yellow to blue index). In the case of CBB staining, the b value is expected to change greatly.

  17) (Evaluation criteria) ΔE is obtained. It can be evaluated that the smaller the ΔE, the smaller the change from the white state = high pellicle removal effect.

ΔE = [(ΔL ^* ) ² + (Δa ^* ) ² + (Δb ^* ) ² ] ^1/2
Here, ΔL ^* = L value after CBB staining−initial L value Δa ^* and Δb ^* are similarly calculated. The smaller the ΔE value, the smaller the amount of protein attached to the surface to be tested, that is, the higher the pellicle decomposition effect, and the results were symbolized as follows.
×: No pellicle decomposition effect (coloring equivalent to positive control)
Δ: Pellicle decomposition effect is weak. ○: Pellicle decomposition effect is present. ◎: Pellicle decomposition effect is strong (coloring to the same extent as negative control).

(10.4.3 Test with bovine enamel slice (basically the same as Hap disc))
1) Saliva is collected by using a saliva collection tube (SARESTED Salivet series, Sarikids series, Sarisoft series, etc. Here, Sarivet 51.1534 is used).

  2) Centrifuge at 2500 rpm for 4 minutes to unite the saliva collected in the tube once.

  3) Two pieces of bovine enamel slices (polished with sandpaper of about # 800) are put into two 50 ml tubes.

  4) Put 4 ml of saliva into a tube containing bovine teeth and confirm that the section is immersed.

  5) Warm the shaking incubator to 37 ° C and incubate overnight (16 hours) with 4 tubes in strong shaking.

  6) The rest of the operation is the same as for the Hap disc. The amount of physiological saline or decoloring solution used for washing is adjusted and increased as appropriate.

(10.5 Evaluation of tea-modified pellicle decomposition effect)
(Test with 10.5.1 Hap powder)
1) Saliva is collected by using a saliva collection tube (SARESTED Salivet series, Sarikids series, Sarisoft series, etc. Here, Sarivet 51.1534 is used).

  2) Centrifuge at 2500 rpm for 4 minutes to unite the saliva collected in the tube once.

  3) Weigh 10 mg of Hap powder into a 2 ml tube.

  4) Add 1 ml of saliva to a tube containing Hap powder and vortex (fully agitate). Add 1 ml of saline to the negative control and vortex in the same way.

  5) Warm the shaking incubator to 37 ° C and incubate overnight (16 hours) with 4 tubes in strong shaking.

  6) Remove from the incubator and centrifuge at 13,000 rpm for 5 minutes to remove the supernatant.

  7) Add 200 μl of physiological saline, and tap to disperse the powder in the physiological saline.

  8) Centrifuge at 13,000 rpm for 5 minutes to remove the supernatant.

  9) Add 1 ml of black tea (100 ml of hot water for 1 bag of tea pack, extract to 5 minutes after extraction for 5 minutes) to the tube, and tap to disperse the powder. Add black tea to the negative control in the same way.

  10) Warm the shaking incubator to 37 ° C and incubate for 1 hour with 9 tubes and strong shaking.

  11) Remove from the incubator and centrifuge at 13,000 rpm for 5 minutes to remove the supernatant.

  12) Add 200 μl of physiological saline, and tap to disperse the powder in the physiological saline.

  13) Centrifuge at 13,000 rpm for 5 minutes to remove the supernatant.

  14) Add a solution of a substance whose effect is to be confirmed, such as an enzyme solution, to the tube, and incubate with strong shaking for 20 minutes in a 37 ° C shaking incubator. Saline is added to the positive and negative controls. * Dry rice bran extract A aqueous solution, purified enzyme derived from Aspergillus genus (eg Amano Enzyme “Protease M Amano SD”, Kikkoman “Tannase KT05”) aqueous solution of protein digestion power 100 U / ml, tannase activity 14.8 U / ml Served in.

  15) Remove from the incubator and centrifuge at 13,000 rpm for 5 minutes to remove the supernatant.

  16) Add 100 μl of physiological saline, and tap to disperse the powder in the physiological saline.

  17) After sonication (15 minutes), centrifuge at 13,000 rpm for 5 minutes, and quantitate protein in the supernatant.

  18) Since the supernatant is also colored and normal quantification may not be possible, prepare a sample blank. Measure absorbance at 595 nm in a 96-well plate with 10 μl of supernatant and 200 μl of distilled water.

  19) (Confirmation of black tea concentration) Add 50 μl of the supernatant to a 96-well plate, and add 125 μl of distilled water to make a total of 150 μl. Measure absorbance at 420 nm. Alternatively, 405, 450, and 430 nm are acceptable.

20) (Evaluation criteria) It was evaluated that the pellicle removal effect was lower as the amount of detected protein was larger, and the effect was higher as the amount was smaller. Further, the amount of black tea pigment was evaluated to be higher as the absorbance was higher, that is, the effect of decomposing the modified pellicle was lower, and as the absorbance was lower, the effect was lower, ie, the effect of decomposing the modified pellicle was higher. The two results were combined and symbolized as follows.
×: No tea-denatured pellicle decomposition effect (coloring and protein amount comparable to positive control)
Δ: Tea-modified pellicle decomposition effect weak ○: Tea-modified pellicle decomposition effect ◎: Tea-modified pellicle decomposition effect strong (coloring and protein amount comparable to negative control).

(Test with 10.5.2 Hap disc)
1) Saliva is collected by using a saliva collection tube (SARESTED Salivet series, Sarikids series, Sarisoft series, etc. Here, Sarivet 51.1534 is used).

  2) Centrifuge at 2500 rpm for 4 minutes to unite the saliva collected in the tube once.

  3) Place the Hap discs one by one in a 2 ml tube.

  4) Add 1 ml of saliva to the tube containing the Hap disk, and add physiological saline to the negative control.

  5) Warm the shaking incubator to 37 ° C and incubate overnight (16 hours) with 4 tubes in strong shaking.

  6) Remove from the incubator and remove saliva (or saline).

  7) Put 200 μl of physiological saline into the tube and rinse lightly to remove the physiological saline. Repeat the same operation once more.

  8) Add 1 ml of black tea (100 ml of hot water for 1 bag of tea pack, extract for 5 minutes and then cool to about 37 ° C.) to the tube.

  9) Warm the shaking incubator to 37 ° C, and incubate for 1 hour with the tube 8 and shaking vigorously.

  10) Remove from the incubator and remove the tea liquor.

  11) Add a solution of a substance whose effect is to be confirmed, such as an enzyme solution, to the tube and incubate with strong shaking for 20 minutes in a shaking incubator at 37 ° C. Saline is added to the positive and negative controls. * Dry rice bran extract A aqueous solution Aspergillus purified enzyme (eg; Amano Enzyme “Protease M Amano SD”, Kikkoman “Tannase KT05”) aqueous solution, protein digestion power 100 U / ml, tannase activity 14.8 U / ml Served in.

  12) Remove from the incubator and remove the enzyme solution.

  13) Add 200 μl of normal saline, lightly rinse and remove.

  14) Remove the disc from the tube and take a picture with a digital camera. At that time, CasMatch is reflected.

  15) (Image processing) Correction is made using CasMatch in the photograph, and then the Lab value is confirmed (using Adobe Photoshop). The larger the L value (brightness), the brighter. a value (red to green index). b value (yellow to blue index). In the case of black tea dyeing, the a value is expected to change greatly.

16) (Evaluation criteria) ΔE is obtained. It can be evaluated that the smaller the ΔE, the smaller the change from the white state = high pellicle removal effect.
ΔE = [(ΔL ^* ) ² + (Δa ^* ) ² + (Δb ^* ) ² ] ^1/2
Here, ΔL ^* = L value after the black tea dyeing enzyme reaction−initial value (before black tea dyeing) L value
Δa ^* and Δb ^* are similarly calculated.
The smaller the ΔE value, the smaller the amount of black tea-modified protein attached to the surface to be tested, that is, the higher the black tea-modified pellicle decomposition effect, and the results were symbolized as follows.
×: No tea-modified pellicle decomposition effect (coloring equivalent to positive control)
△: Tea-modified pellicle decomposition effect weak ○: Tea-modified pellicle decomposition effect ◎: Tea-modified pellicle decomposition effect strong (coloring comparable to negative control)
(10.5.3 Test on bovine enamel slice (basically the same as Hap disc))
1) Saliva is collected by using a saliva collection tube (SARESTED Salivet series, Sarikids series, Sarisoft series, etc. Here, Sarivet 51.1534 is used).

  2) Centrifuge at 2500 rpm for 4 minutes to unite the saliva collected in the tube once.

  3) Place two pieces of bovine enamel slices in a 50 ml tube.

  4) Put 4 ml of saliva into a tube containing bovine teeth and confirm that the section is immersed.

  5) Warm the shaking incubator to 37 ° C and incubate overnight (16 hours) with 4 tubes in strong shaking.

  6) The rest of the operation is the same as for the Hap disc. The amount of the enzyme solution and the amount of physiological saline used for washing are appropriately adjusted and increased.

(10.6 Evaluation of the effect of inhibiting the deposition of black tea on the pellicle)
(Test with 10.6.1 Hap powder)
1) Saliva is collected by using a saliva collection tube (SARESTED Salivet series, Sarikids series, Sarisoft series, etc. Here, Sarivet 51.1534 is used).

  2) Centrifuge at 2500 rpm for 4 minutes to unite the saliva collected in the tube once.

  3) Weigh 10 mg of Hap powder into a 2 ml tube.

  4) Add 1 ml of saliva to a tube containing Hap powder and vortex (fully agitate). Add 1 ml of saline to the negative control and vortex in the same way.

  5) Warm the shaking incubator to 37 ° C and incubate overnight (16 hours) with 4 tubes in strong shaking.

  6) Remove from the incubator and centrifuge at 13,000 rpm for 5 minutes to remove the supernatant.

  7) Add 200 μl of physiological saline, and tap to disperse the powder in the physiological saline.

  8) Centrifuge at 13,000 rpm for 5 minutes to remove the supernatant.

  9) Dissolve (disperse) substances, such as enzymes, in tea (100 ml of hot water for 1 pack of tea packs, extract for 5 minutes and then cool to about 37 ° C), add them to the tube, and shake at 37 ° C Incubate with strong shaking for 1 hour in incubator. Add tea to the positive and negative controls. * Dry rice bran extract A aqueous solution, purified enzyme derived from Aspergillus genus (eg Amano Enzyme “Protease M Amano SD”, Kikkoman “Tannase KT05”) aqueous solution of protein digestion power 100 U / ml, tannase activity 14.8 U / ml Served in.

  10) Remove from the incubator and centrifuge at 13,000 rpm for 5 minutes to remove the black tea enzyme solution.

  11) Add 200 μl of physiological saline, and tap to disperse the powder in the physiological saline.

  12) Centrifuge at 13,000 rpm for 5 minutes to remove the supernatant.

  13) Add 100 μl of physiological saline, and tap to disperse the powder in the physiological saline.

  14) After sonication (15 minutes), centrifuge at 13,000 rpm for 5 minutes, and quantitate protein in the supernatant.

  15) Since the supernatant is also colored and normal quantification may not be possible, prepare a sample blank. Measure absorbance at 595 nm in a 96-well plate with 10 μl of supernatant and 200 μl of distilled water.

  16) (Confirmation of black tea concentration) Add 50 μl of the supernatant to a 96-well plate and add 125 μl of distilled water to make a total of 150 μl. Measure absorbance at 420 nm. Alternatively, 405, 450, and 430 nm are acceptable.

17) It was evaluated that the greater the amount of protein detected, the lower the pellicle removal effect, and the smaller the amount, the higher the effect. Also, the amount of black tea pigment was higher as the absorbance was higher, that is, the amount of black tea deposited on the pellicle was lower, and the lower the absorbance was, the lower the black tea deposit was evaluated. The two results were combined and symbolized as follows:
×: No effect of suppressing black tea deposition on the pellicle (coloring and protein amount comparable to the positive control)
Δ: Black tea deposition inhibitory effect on pellicle is weak ○: Black tea deposition inhibitory effect on pellicle is good ◎: Black tea deposition inhibitory effect on pellicle is strong (coloring and protein amount comparable to negative control).

(Test with 10.6.2 Hap disc)
1) Saliva is collected by using a saliva collection tube (SARESTED Salivet series, Sarikids series, Sarisoft series, etc. Here, Sarivet 51.1534 is used).

  2) Centrifuge at 2500 rpm for 4 minutes to unite the saliva collected in the tube once.

  3) Place the Hap discs one by one in a 2 ml tube.

  4) Add 1 ml of saliva to the tube containing the Hap disk, and add physiological saline to the negative control.

  5) Warm the shaking incubator to 37 ° C and incubate overnight (16 hours) with 4 tubes in strong shaking.

  6) Remove from the incubator and remove saliva (or saline).

  7) Put 200 μl of physiological saline into the tube and rinse lightly to remove the physiological saline. Repeat the same operation once more.

  8) Dissolve (disperse) substances such as enzymes in tea (100 ml of hot water for 1 pack of tea pack, extract for 5 minutes, then cool to about 37 ° C), add to tube, and shake at 37 ° C Incubate with strong shaking for 1 hour in incubator. Add tea to the positive and negative controls. * Dry rice bran extract A aqueous solution, purified enzyme derived from Aspergillus genus (eg Amano Enzyme “Protease M Amano SD”, Kikkoman “Tannase KT05”) aqueous solution of protein digestion power 100 U / ml, tannase activity 14.8 U / ml Served in.

  9) Remove from the incubator and remove the tea enzyme solution.

  10) Put 200 μl of physiological saline into the tube and rinse lightly to remove the physiological saline. Repeat the same operation once more.

  11) Remove the disc from the tube and take a picture with a digital camera. At that time, CasMatch is reflected.

  12) (Image processing) Correction is performed using CasMatch in the photograph, and then the Lab value is confirmed (using Adobe Photoshop). The larger the L value (brightness), the brighter. a value (red to green index). b value (yellow to blue index). In the case of black tea dyeing, the a value is expected to change greatly.

13) (Evaluation criteria) ΔE is obtained. The smaller the ΔE, the smaller the change from the white state = it can be evaluated that the effect of suppressing the black tea deposition on the pellicle is high.
ΔE = [(ΔL ^* ) ² + (Δa ^* ) ² + (Δb ^* ) ² ] ^1/2
Here, ΔL ^* = L value after the black tea dyeing enzyme reaction−initial value (before black tea dyeing) L value
Δa ^* and Δb ^* are similarly calculated.
It was evaluated that the smaller the ΔE value, the smaller the amount of black tea-modified protein attached to the surface to be tested, that is, the higher the effect of suppressing black tea deposition on the pellicle, and the results were symbolized as follows.
×: No effect of suppressing black tea deposition on the pellicle (coloring equivalent to the positive control)
Δ: Black tea deposition inhibitory effect on pellicle is weak ○: Black tea deposition inhibitory effect on pellicle is good ◎: Black tea deposition inhibitory effect on pellicle is strong (same color as negative control).

(10.6.3 Test with bovine enamel slice)
1) Saliva is collected by using a saliva collection tube (SARESTED Salivet series, Sarikids series, Sarisoft series, etc. Here, Sarivet 51.1534 is used).

  2) Centrifuge at 2500 rpm for 4 minutes to unite the saliva collected in the tube once.

  3) Place two pieces of bovine enamel slices in a 50 ml tube.

  4) Put 4 ml of saliva into a tube containing bovine teeth and confirm that the section is immersed.

  5) Warm the shaking incubator to 37 ° C and incubate overnight (16 hours) with 4 tubes in strong shaking.

  6) The rest of the operation is the same as for the Hap disc. The amount of the enzyme solution and the amount of physiological saline used for washing are appropriately adjusted and increased.

(Production Example 1: Method for producing dried rice bran extract)
Commercially available rice bran (trade name: dried rice cake for sweet sake manufactured by Korsafes Co., Ltd.) is mixed with ion-exchanged water of 3 times the weight at room temperature (about 20 ° C) in the extraction tank. For 2 hours. During stirring, the mixture was slowly and intermittently stirred so as not to crush the wrinkles. The mixture was subjected to solid-liquid separation using a dispensing pump and a separation net, and finally a 60-mesh extract was obtained. It was Brix5 when the Brix sugar content of this extract was measured. The yield of this extract was about 65% by weight of the ion-exchanged water used. Next, this extract solution is put into a stirring tank, and an indigestible dextrin (Fibersol 2 manufactured by Matsutani Chemical Industry Co., Ltd.) having a weight 0.5 times the weight of the raw material koji is added, and at room temperature (about 20 ° C) The mixture was dissolved to obtain a mixed solution. The liquid mixture was dried by spray drying and applied to a shifter to obtain a dried soot extract of 50 mesh pass. The dried koji extract produced in Production Example 1 is referred to as A for convenience. The outline of the manufacturing flow of the dried rice bran extract from this rice bran is shown in FIG.

(Production Comparative Example 1: Method for producing dried rice bran extract)
A dried rice bran extract was obtained in the same manner as in Production Example 1, except that a commercially available rice bran (trade name dried rice bran for shochu made by Korsafes Co., Ltd.) was used. The dried soot extract produced in Production Comparative Example 1 is referred to as B for convenience.

(Production Comparative Example 2: Method for producing dried rice bran extract)
A dried rice bran extract was obtained in the same manner as in Production Example 1, except that a commercially available rice bran (trade name dried rice bran for shochu made by Korsafes Co., Ltd.) was used. The dried soot extract produced in Production Comparative Example 2 is referred to as C.

(Production Comparative Example 3: Method for producing dried rice bran extract)
A dried rice bran extract was obtained in the same manner as in Production Example 1, except that a commercially available rice bran (trade name dried rice bran for shochu made by Korsafes Co., Ltd.) was used. The dried soot extract produced in Production Comparative Example 3 is referred to as D for convenience.

(Production Comparative Example 4: Rice bran frozen pulverized powder)
The same commercially available rice bran as used in Production Example 1 (trade name, dried rice cake for sweet sake manufactured by Korsafes Co., Ltd.) was freeze-dried and then pulverized to obtain a rice bran freeze-ground powder.

(Component analysis value of dried rice bran extract produced in Production Example 1)
The components of the dried rice bran extract produced in Production Example 1 and Production Comparative Examples 1 to 4 were analyzed. The results are shown in Table 3 below.

(Measurement Example 1: Measurement of various enzyme activities of dried rice bran extract and other powders produced in Production Example 1)
Dry rice bran extract produced according to the method described in Production Example 1, dry rice bran extract produced according to the method described in Production Comparative Examples 1 to 3, frozen rice bran produced according to the method described in Production Comparative Example 4 Various enzyme activities of the pulverized powder and commercially available tannase (manufactured by Kikkoman Corporation, tannase KT05 (food additive)) were measured according to the method described in “10. Measuring Method” of the present specification. The results are shown in Table 4 and Table 5 below.

  As a result, it was confirmed that the dried rice bran extract of the present invention has very high tannase activity and acidic protease activity.

(Measurement Example 2: Effect of rice bran in vitro (Hap powder and Hap disc))
Dry rice bran extract A prepared in Production Example 1 (freeze-ground product of rice bran extract; acid protease 5547 U / g, α-amylase 606 U / g, glucoamylase 147 U / g, tannase 1430 U / g) was used. Then, a pellicle decomposition effect verification test was performed according to “10.4. Evaluation of pellicle decomposition effect” in this specification.

  Hydroxyapatite (Hap) powder and a Hap fired disc were shaken overnight (16 hours) in saliva, then immersed in the following three solutions and shaken at 37 ° C. for 20 minutes. The protein remaining in the Hap powder was physically dropped and the amount was quantified. The Hap disk was stained with CBB and then photographed with a digital camera, image analysis was performed, and whitening was evaluated.

<Test solution>
The following test solutions were used:
(1) physiological saline (PBS), (2) dried rice bran extract A solution (protease concentration 100 U / ml), (3) actinidine solution (protease concentration 100 U / ml). As a negative control, the amount of protein adhesion was measured after being immersed in physiological saline instead of saliva, and the amount of adhesion was subtracted from the amount of protein adhesion in each experiment. The results are shown in Table 6 below and FIG. As a result, the pellicle was most removed when it was treated with the dried rice bran extract A solution.

  The result about the effect of the dried rice bran extract A in vitro when using a Hap disk is shown in FIG. As a result, it was confirmed that the amount of protein adhered was the highest when PBS was used, and the amount of protein adhered was the smallest when dry rice bran extract A was used.

(Example 1A: Confirmation of pellicle decomposition effect)
Using hydroxyapatite powder, an experiment for confirming the pellicle decomposition effect was performed based on "10.4 Evaluation of pellicle decomposition effect" in this specification.

  The results are shown in Table 7A below and FIG. FIG. 4 shows the results of verification using hydroxyapatite powder.

  As a result, it was confirmed that the dried koji extract of the production example of the present invention has a significant pellicle decomposition effect. This pellicle degradation effect is superior to purified commercial proteases with the same protease activity.

(Example 1B: Confirmation of pellicle decomposition effect)
Using the bovine enamel slice, an experiment for confirming the pellicle decomposition effect is performed based on “10.4 Evaluation of pellicle decomposition effect” in this specification.

  The results are shown in Table 7B below.

  As a result, it is confirmed that the dried koji extract of the production example of the present invention has a significant pellicle decomposition effect.

(Example 2: Confirmation of tea-modified pellicle decomposition effect)
Using the hydroxyapatite powder and the bovine enamel slice, an experiment for confirming the pellicle decomposition effect is performed based on “10.5 Evaluation of tea-modified pellicle decomposition effect” in this specification.

  The results are shown in Table 8 below:

  As a result, it is confirmed that the dried koji extract of the present invention exhibits an excellent black tea deposition inhibiting effect. This tea-denatured pellicle degradation effect is superior to the combination of purified commercial protease and tannase, which have the same protease activity and tannase activity.

(Example 3: Confirmation of black tea deposition inhibitory effect)
Using the hydroxyapatite powder and the bovine enamel slice, an experiment for confirming the black tea deposition inhibiting effect is performed based on “10.5 Evaluation of black tea deposition inhibiting effect on pellicle” in the present specification.

  The results are shown in Table 9 below:

  As a result, it is confirmed that the dried koji extract of the present invention exhibits an excellent black tea deposition inhibiting effect. This black tea deposition inhibitory effect is superior to a combination of a commercially available purified protease and tannase having the same protease activity and tannase activity.

(Example 4: Production of gum containing dried rice bran extract)
After kneading 25 parts by weight of gum base, 72 parts by weight of palatinose, 2 parts by weight of reduced mizuame and 1 part by weight of dried rice bran extract A produced in Production Example 1, a molded board gum was prepared. The weight of the obtained board gum was 2.0 g. This board gum had a carious saccharide content of 0.15% by weight and a non-cariogenic saccharide content of 74.8% by weight.

  As a control, a plate gum using the dried rice bran extract produced in any of Production Comparative Examples 1 to 4 instead of the dried rice bran extract A in Production Example 1 was also prepared.

(Example 5: Production of tablet containing dried rice bran extract)
After mixing 99 parts by weight of reduced palatinose and 1 part by weight of the dried rice bran extract A produced in Production Example 1, using a circular tableting mold having a diameter of 13 mm, the suction depth was 12 mm and the center tableting pressure was 2 t. The mixture was tableted under conditions. The weight of one tableted product (that is, tablet) obtained was 1.0 g. This tablet had a carious saccharide content of 0.15% by weight and a non-cariogenic saccharide content of 99.8% by weight.

  As a control, a tablet using the dried rice bran extract produced in any of Production Comparative Examples 1 to 4 instead of the dried rice bran extract A in Production Example 1 was also prepared.

(Example 6: Confirmation of preference in dry rice bran extract)
Five panelists are allowed to taste the dry rice bran extract A1% blended gum, tablets, and rice bran frozen ground powder 1% blended gum and tablets produced in Example 4 or 5, and evaluate the taste. The evaluation criteria are set as follows and the average of 5 persons is shown. The results are shown in Table 10 below.

◎… Does not taste at all 麹 / starch ○… Slightly tastes 麹 / starch △… Slightly tastes 麹 / starch ×… Only tastes like 麹 / starch

(Example 7: Confirmation of superiority in use feeling of dried rice bran extract)
Evaluate in terms of usability by having 5 panelists taste the dry rice bran extract 1% blended gum, tablets and 1% rice bran frozen ground powder 1% blended gum and tablets produced in Example 4 or 5. Do. The evaluation criteria are set as follows and the average of 5 persons is shown. The results are shown in Table 11 below.

◎… No unpleasant roughness / stickiness ○… Slightly unpleasant roughness / stickiness △… Some of unpleasant roughness / stickiness ×… Severe roughness / stickiness is too bad to eat

(Example 8: Confirmation of human experience of whitening effect of tooth by dry rice bran extract)
Let 5 subjects ingest 2% 20 minutes of dry rice bran extract 1% mixed gum or dry rice bran extract non-blended gum produced in Example 4 or 5 after each meal in the morning, noon and night. Continue for 10 days. In addition, during this period, tea is ingested three times a day. The change of the color of one's own teeth when compared with before continuous intake after completion is evaluated according to the following criteria, and an average of 5 people is shown. The results are shown in Table 12 below.

◎… I feel that it is clearly whiter than before continuous intake ○… I feel a little whiter than before continuous intake △… I feel almost the same as before continuous intake ×… I feel that whiteness is lost than before continuous intake

(Example 9: Confirmation of tooth whitening effect by dry rice bran extract by human intraoral photograph)
Five subjects ingested 2% of dried rice bran extract 1% gum or non-dried rice bran extract blended gum prepared in Example 4 or 5 after each meal in the morning, noon and night. For 10 days. In addition, during this period, tea is ingested three times a day. Photographs in the oral cavity are taken before and after the continuous ingestion, and the color tone of the teeth is evaluated by Adobe Photoshop in the Lab color system. ΔE is calculated by the following equation.
ΔE = [(ΔL ^* ) ² + (Δa ^* ) ² + (Δb ^* ) ² ] ^1/2
At this time, ΔL ^* = (L value after continuous intake for 10 days) − (L value before continuous intake)
It calculates similarly about (DELTA) a ^* and (DELTA ⁾ b ^* . During the period, ΔE rises due to the continued intake of black tea, but if the test food has a whitening effect, the rise is expected to be small. The evaluation is classified into the following stages based on the value of ΔE. The results are shown in Table 13 below.

◎… ΔE <0.1
○ ... 0.1 ≦ ΔE <1
Δ ... 1 ≦ ΔE <10
× ... 10 ≦ ΔE

(Example 10: Measurement of tongue coating removal effect and halitosis suppression effect)
The following method was used for 5 healthy subjects except that the tongue coating removal effect of the dry rice bran extract 1% tablet or non-dried rice bran extract formulation tablet produced in Example 4 or 5 was required. Evaluate with:
(1) First, photograph the subject's tongue back and evaluate the tongue coating score by visual inspection:
(I) Photographing method The subject's tongue back is photographed using a digital camera (FinePix S602, Fuji Film) equipped with a ring flash (14RDX, YUZO). In this method, a photograph without a shadow is taken. A color chart for color tone correction (CasMatch, Dai Nippon Printing Co., Ltd.) is photographed next to the subject, and the brightness of the photograph is corrected using this.

(Ii) Evaluation method by visual inspection Evaluation of the amount of tongue coating by visual inspection is performed by Moriya et al. (Toshiki Moriya, Mitsuo Kishi, Fumie Aizawa et al. 2002) and adding the thickness of tongue coating to the method. That is, the photographed tongue back is divided into four areas, and the amount of tongue coating attached to each area is evaluated as (tongue coating attachment score) × (tongue coating thickness score), and the total of these is used as tongue coating score. . The evaluation is an average value of three evaluators. The tongue coating adhesion score and tongue coating thickness score are both 4 grades of 0-3.

1) Divide the back of the tongue into four areas;
2) The amount of tongue coating attached is evaluated from the area and thickness of tongue coating attached to each area. Tongue coating amount = (tongue coating attachment area score) x (tongue coating thickness score);
3) The total score of each area is the tongue coating score.

  The criteria for tongue coating area score are as follows:

  The criteria for tongue thickness ratings are as follows:

  (2) Have the subject lick the tablet so as not to chew and swallow. As a result, it takes about 7 minutes to lick.

  (3) Immediately after finishing licking, take a picture of the back of the tongue in the same manner as in (1), and evaluate the tongue coating score by visual inspection.

  (4) For each subject, compare the tongue coating scores of (1) and (3). The results for tongue coating removal effect are shown in Table 16 below.

Further, when evaluating the tongue coating removal effect, the bad breath removal effect is also measured by the following method. Attention was paid to H ₂ S and CH ₃ SH as volatile sulfide compounds (VSC). This is because H ₂ S and CH ₃ SH have a high contribution to bad breath.

Specifically, before taking a tablet, one tablet was licked so that it was not chewed and swallowed (thus, it took about 7 minutes to lick), 20 minutes later, and then another tablet After licking so as not to chew and swallow, 20 minutes later, the concentration of H ₂ S and CH ₃ SH is measured using Oral Chroma (Abilit). The subject is instructed to breathe nasally for 30 seconds while holding the 10 ml syringe so that the mouth does not leak after one deep breath. Thereafter, 10 ml of mouth air is collected, and 5 ml thereof is injected into oral chroma (Abilit). The concentration of H ₂ S and CH ₃ SH is calculated as the weight (ng / 10 ml) in 10 ml of mouth air using the dedicated analysis software of a personal computer connected to the oral chroma. This evaluation is performed on 5 healthy subjects except that tongue coating removal is required. The results for the bad breath suppression effect are shown in Table 17 below.

  As a result, it is confirmed that the dried rice bran extract of the present invention has not only a stain preventing effect and a removing effect, but also a tongue coating removing effect and a bad breath suppressing effect.

  As mentioned above, although this invention has been illustrated using preferable embodiment of this invention, this invention should not be limited and limited to this embodiment. It is understood that the scope of the present invention should be construed only by the claims. It is understood that those skilled in the art can implement an equivalent range based on the description of the present invention and the common general technical knowledge from the description of specific preferred embodiments of the present invention. Patents, patent applications, and documents cited herein should be incorporated by reference in their entirety, as if the contents themselves were specifically described herein. Understood.

  The food and oral composition of the present invention are useful for removing colored stains on teeth and preventing deposition. Since the food and oral cavity composition of the present invention have a very low content of carious saccharides, the use of the food and oral cavity composition has little or no adverse effect on oral dental caries.

  The dried koji extract of the present invention is cheaper than a plurality of combinations of purified enzymes and is easily prepared.

Claims (23)

A dried koji extract,
The dry cocoon extract comprises a dry product of a mixture comprising cocoon extract and a non-cariogenic excipient,
The dry koji extract has protease activity and tannase activity.   2. The dry cocoon extract according to claim 1, wherein the non-cariogenic excipient is selected from the group consisting of sugar alcohols and non-cariogenic saccharides.   The dry koji extract according to claim 2, wherein the total content of sugar alcohol and non-cariogenic sugar in the dry koji extract is 20 wt% to 95 wt%.   The dry koji extract according to any one of claims 1 to 3, wherein a moisture content of the dry koji extract is 0.001 wt% to 5 wt%.   The dry koji extract according to any one of claims 1 to 4, wherein a content of carious saccharide in the dry koji extract is 20% by weight or less.   The dry koji extract according to any one of claims 1 to 5, wherein the protease activity is 500 U / g or more.   The dry koji extract according to any one of claims 1 to 6, wherein the tannase activity is 50 U / g or more.   The non-cariogenic excipient is selected from the group consisting of indigestible dextrin, palatinose, trehalose, lactitol, reduced palatinose, maltitol, erythritol, sorbitol, xylitol or mannitol. The dried koji extract according to item.   The dry koji extract according to any one of claims 1 to 8, wherein the dry koji extract is produced from rice bran.   The foodstuff containing the dry koji extract of any one of Claims 1-9, and the content of a carious saccharide is 0.5 weight% or less.   The food according to claim 10, which is used for removing tongue coating.   The food according to claim 10 or 11, which is a gum, candy or tablet.   The foodstuff of any one of Claims 10-12 whose protease activity is 100 U / g or more.   The food according to any one of claims 10 to 13, wherein the tannase activity is 10 U / g or more.   The composition for oral cavity containing the dry candy extract of any one of Claims 1-9, and the content of a carious saccharide is 0.5 weight% or less.   The composition for oral cavity of Claim 15 which is a dentifrice or a mouthwash.   The composition for oral cavity according to claim 15, which is a troche or gel.   The composition for oral cavity of any one of Claims 15-17 whose protease activity is 50 U / g or more.   The composition for oral cavity of any one of Claims 15-18 whose tannase activity is 5 U / g or more. A method for producing a dried koji extract according to claim 1,
Mixing the straw with water at 0 ° C. to 25 ° C. for 30 minutes to 3 hours;
Separating the extract from the mixture;
A method comprising the steps of mixing the extract and a non-cariogenic excipient to obtain an extract excipient mixture; and drying the extract excipient mixture to obtain a dry anther extract.   21. The method of claim 20, wherein the non-cariogenic excipient is a sugar alcohol or a non-cariogenic saccharide.   The method according to claim 20 or 21, wherein the rice bran is rice bran.   The method according to any one of claims 20 to 22, wherein the drying is freeze-drying or spray-drying.