JP2007106683A - Fat accumulation inhibitor and lipolysis accelerator - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a fat accumulation inhibitor and lipolysis accelerator which have the action of inhibiting new accumulation of the body fat and reducing the body fat already accumulated and is suitable for use as an obesity preventive agent or an obesity ameliorating agent. <P>SOLUTION: The fat accumulation inhibitor and lipolysis accelerator contain a hydrolyzate of a wheat protein and is suitably used as an obesity preventing agent or an obesity ameliorating agent. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

  The present invention relates to a fat accumulation inhibitor and a lipolysis promoter comprising a wheat protein hydrolyzate. Specifically, it contains a hydrolyzate of wheat protein, has both fat accumulation inhibiting action and lipolysis promoting action, and is suitable as an obesity preventive or obesity improving agent, and fat accumulation inhibiting and lipolysis promoting agent, and said fat accumulation inhibiting And a food or drink containing a lipolysis promoter.

So far, slimming agents and lipid metabolism improving agents containing various substances as active ingredients have been developed.
For example, catechins contained in tea and the like are known to have a burning promoting effect on body fat and dietary fat, and a diet using a tea extract containing a high concentration of catechins and a concentrated liquid thereof. Beverages have been proposed (Patent Document 1). In addition, amino acids such as lysine, proline, alanine, and arginine are known to have a lipolytic action, and slimming agents containing these have been proposed (Patent Document 2). However, these ingredients have peculiar tastes and odors such as bitterness and sardine, and it has been difficult to ingest these ingredients until the above effects are fully exhibited.

  In addition, it has been found that a peptide containing a large amount of L-glutamine (hereinafter also referred to as glutamine peptide) exhibits a serum lipid lowering action for hyperlipidemia, and this glutamine peptide is used as a prophylactic agent for hyperlipidemia and / or Alternatively, use as a therapeutic agent has been proposed (Patent Document 3). In addition, it is known that an enzymatic degradation product of wheat red straw improves lipid metabolism, and it has been proposed to add this enzymatic degradation product to pharmaceuticals, health foods, and various foods and drinks (Patent Document 4). However, it has been clarified that these components have the effect of lowering blood lipid levels and lipid levels in the liver, but whether or not new fat accumulation in fat cells is suppressed or has already accumulated. There is no indication that it can reduce body fat. Therefore, it has been completely unknown whether these components have an action of suppressing accumulation and decomposition of body fat and are effective in preventing or improving obesity.

  Therefore, it suppresses new accumulation of body fat and reduces the accumulated body fat, has an excellent obesity-preventing action or obesity-improving action, has no side effects, is highly safe, and is easily consumed. There is still a great demand for fat accumulation inhibitors and lipolysis promoters, and foods and drinks containing these agents, which are possible and can be continuously ingested for a long period of time, and are suitable as obesity-preventing or obesity-improving agents. high.

  On the other hand, apart from these, there is an attempt to obtain a slimming effect by inhibiting the differentiation of preadipocytes into adipocytes, and it has cell differentiation inhibiting activity such as coffee bean extract and globin proteolysate The material is known. However, in recent years, the importance of adipocytes as secretory organs such as adipocytokines has been reviewed, and it is considered that intake of components having a strong cell differentiation inhibitory action is not preferable. Furthermore, the decrease in the number of fat cells resulting in a decrease in the place for storing triglycerides (TG) in blood is thought to lead to an increase in the blood concentration of TG.

Therefore, agents and foods and drinks for the prevention and improvement of obesity, while maintaining the number of fat cells, suppress the accumulation of new fat in fat cells and have already accumulated fat in fat cells. It is desirable to have the effect of reducing the amount.
JP 2002-326932 A JP 2003-119133 A WO03 / 074071 JP-A-8-81390

  The present invention provides an agent for inhibiting fat accumulation and promoting lipolysis, which has an action of suppressing new accumulation of body fat and reducing the accumulated body fat, and is suitable as an obesity preventing agent or an obesity improving agent. The challenge is to do.

  In addition, the present invention comprises the above-described fat accumulation inhibitor and lipolysis promoter, has an excellent obesity-preventing action or obesity-improving action, has no side effects, is highly safe, and can be easily ingested. It is also an issue to provide foods and drinks that can be easily taken for a long time.

  As a result of intensive studies to solve the above problems, the present inventors have found that a peptide mixture obtained by hydrolyzing wheat protein has an excellent fat accumulation-inhibiting action and lipolysis-promoting action. It came to be completed.

That is, the present invention relates to the following matters.
The fat accumulation inhibiting and lipolysis promoter according to the present invention is characterized by containing a hydrolyzate of wheat protein.

In the present invention, the wheat protein hydrolyzate preferably contains a peptide mixture having a molecular weight of 200 to 2,000 Da in an amount of 50 to 90% by mass.
Furthermore, the fat accumulation inhibitor and lipolysis promoter of the present invention do not have a cell differentiation inhibitory activity from preadipocytes to adipocytes.

The agent for inhibiting fat accumulation and promoting lipolysis of the present invention is suitably used as an obesity preventing agent or an obesity improving agent.
Moreover, the food / beverage products which concern on this invention are characterized by including the said fat accumulation suppression and a lipolysis promoter.

  According to the present invention, the new accumulation of body fat is suppressed and the already accumulated body fat is reduced, and an excellent obesity preventing action or obesity improving action is achieved, and there is no worry about side effects and the safety is high. In addition, it is possible to provide a fat accumulation inhibitor and a lipolysis promoter, an obesity preventive agent, an obesity improving agent, and a food and drink, which have a good flavor and can be taken easily and can be taken continuously for a long period of time.

  The fat accumulation inhibiting and lipolysis promoting agent of the present invention has an excellent fat accumulation inhibiting action and lipolysis promoting action, and can remarkably suppress new accumulation of body fat and significantly reduce accumulated fat. In addition, even if taken for a long time, there is no worry about side effects, and it is highly safe and has a good flavor. Furthermore, since the differentiation from preadipocytes to adipocytes is not inhibited, the function of adipocytes as a secretory organ is not impaired.

  Therefore, the fat accumulation inhibitor and lipolysis promoter, obesity preventive agent, obesity improving agent, and food and drink of the present invention can be ingested easily and continuously for a long time in daily life. In addition to the prevention and / or treatment of obesity, body fat loss, slimming, and body can be remarkably inhibited from newly accumulating body fat and can significantly degrade already accumulated body fat. It is also useful for preventing fat accumulation.

Hereinafter, preferred embodiments of the present invention will be specifically described.
The wheat protein used in the present invention contains gluten as a main component, but may contain other proteins known to be contained in wheat such as albumin, globulin, glutenin, gliadin, Furthermore, impurities such as starch and fiber may be inevitably contained. In the present invention, wheat protein that has been subjected to chemical treatment or biological treatment with enzymes or the like in advance to reduce molecular weight or that has increased affinity with protease can be used as wheat protein. .

  The wheat protein hydrolyzate used in the present invention can be obtained by hydrolyzing the above wheat protein, and usually contains a peptide mixture having a molecular weight distribution of about 10,000 Da or less as a main component. Non-peptide components such as water-soluble polysaccharides derived from impurities contained in the wheat protein used may be contained in an amount of 20% by mass or less. The wheat protein hydrolyzate preferably contains a peptide mixture having a molecular weight of 200 to 2,000 Da in an amount of 50 to 90% by mass, more preferably 55 to 85% by mass.

Here, the molecular weight is measured by size exclusion high performance liquid chromatography, and details of measurement conditions and the like are as described in Examples described later.
When the hydrolyzate of wheat protein contains a peptide mixture having a specific molecular weight within the above range, it is highly soluble in water and can be suitably used not only as a solid form but also as a liquid form. Moreover, since there is no taste and smell peculiar to an amino acid, and flavor is good, it is preferable.

  As a method for hydrolyzing wheat protein, a known hydrolysis method can be appropriately employed. Specifically, (1) a method of hydrolyzing using a protease, (2) a method of hydrolyzing using a protease and amylase in combination, (3) a method of hydrolyzing using an acid, and the like.

(1) Examples of the method for hydrolyzing wheat protein using protease include the method described in Japanese Patent No. 3088491.
Specifically, when a wheat protein is hydrolyzed using a protease, a hydrolyzate can be generated by the action of one or more kinds of proteases. And a method using a neutral protease is preferable in that a hydrolyzate of wheat protein which is a target product can be obtained in a high yield.

  When acidic protease and neutral protease are used, acidic protease and neutral protease may be applied to the hydrolysis treatment of wheat protein at the same time. However, after treatment with acidic protease, the acidic protease is deactivated and then neutralized. It is preferable to treat with a functional protease since the dispersibility of the wheat protein is improved in an acidic solution and the hydrolysis reaction efficiency is increased.

When hydrolyzing wheat protein using a protease, it is necessary to adjust appropriately depending on the type of protease to be used. Usually, the concentration of wheat protein in an aqueous medium is 1.0.
It is good to adjust to -20% by mass.

  As the acid protease, pepsin; acid protease derived from Aspergillus or aspartic protease derived from microorganisms such as penicillium-derived aspartic protease; Among them, it is preferable to use an acidic protease derived from pepsin or koji mold because a hydrolyzate of the target wheat protein can be obtained in a high yield.

  One type of acidic protease may be used alone, or a plurality of types may be used in combination as long as acidic proteases do not adversely affect each other. In addition, the acidic protease may be used in a free state or may be used in any case. In either case, the acidic protease is used at a rate of about 5,000 to 250,000 units per 100 g of dried wheat protein. It is good. In the present specification, the protease activity (unit) was determined by using an 1% Hammerstein casein solution manufactured by Merck & Co., USA, as a substrate, and modified by Anson-Hagiwara (edited by Shiro Akahori, “Enzyme Research Method,” Volume 2, The reaction was performed at 30 ° C. for 30 minutes, and the amount of enzyme required to release 1 μg of tyrosine equivalent per minute was defined as 1 unit.

When processing using acidic protease, it is usually good to carry out until the degree of hydrolysis of wheat protein reaches 30% to 60%.
For example, when pepsin or koji mold-derived acidic protease is used as the acidic protease, the pH of the aqueous medium containing wheat protein is adjusted to about 1.5 to 6.0, and the reaction is performed at a temperature of about 30 to 70 ° C. for about 2 to 25 hours. Then, the degree of hydrolysis becomes 30 to 60%. At that time, the acidic protease is deactivated to stop hydrolysis. The inactivation of the acidic protease can be performed by a method in which the pH of the solution is higher than the active pH of the acidic protease or the temperature is increased (usually about 70 to 90 ° C.).

In the present specification, the degree of hydrolysis of wheat protein means the ratio of protein mass dissolved in 0.75M trichloroacetic acid (TCA) in the total protein mass (dissolution rate in TCA), Is required. Hydrolysis degree (%) = [(0.75M TCA dissolved protein mass) / (total protein mass)] × 100
When further processing is performed using a neutral protease, thermolysin, trypsin, chymotrypsin, pronase, subtilisin, esperase, and the like can be used as the neutral protease. Among them, it is preferable to use thermolysin or subtilisin because a hydrolyzate of the target wheat protein can be obtained in high yield.

  Neutral proteases may be used alone or in combination as long as the neutral proteases do not adversely affect each other. In addition, the neutral protease may be used in a free state or may be used after being immobilized. In either case, the neutral protease is in a ratio of about 10,000 to 500,000 units per 100 g of dried wheat protein. It is good to use in. The treatment with the neutral protease is preferably performed until the degree of hydrolysis of the wheat protein reaches 60 to 90%.

  For example, when thermolysin is used as a neutral protease, the pH of an aqueous medium containing wheat protein is adjusted to about 7.0 to 8.0, and the reaction is performed at a temperature of about 30 to 70 ° C. for about 1 to 25 hours. Since the degree becomes 60 to 90%, thermolysin is preferably deactivated at that time. The neutral protease can be inactivated by a method in which the pH of the solution is lower than the active pH of the neutral protease or the temperature is increased (usually about 80 to 100 ° C.).

(2) As a method of hydrolyzing using protease and amylase in combination, for example, the method described in Japanese Patent No. 2985193 can be mentioned.
When wheat protein is hydrolyzed with protease and amylase in combination, amylase degrades starch and fiber present as impurities in wheat protein. It can be obtained with high purity and high yield.

The protease treatment may be performed according to the above method, and the protease treatment and amylase treatment may be performed simultaneously or separately. When performing separately, the amylase treatment may be performed after the protease treatment first, or the protease treatment may be performed after the amylase treatment. The amylase treatment can be carried out simultaneously with the protease treatment or separately, or can be carried out by appropriately selecting conditions such as optimum pH and temperature according to the type of protease used, the type of amylase, and the like.

  When hydrolyzing wheat protein using a protease and amylase in combination, it is necessary to adjust appropriately depending on the type of protease used and the type of amylase. Usually, the concentration of wheat protein in an aqueous medium is 1.0 to 20%. It is better to adjust to mass%.

As the protease, those described above can be used.
As the amylase, α-amylase, β-amylase, glucoamylase, isoamylase, oligosaccharide-producing amylase and the like can be used. Among these, it is preferable to use α-amylase because a hydrolyzate of the target wheat protein can be obtained in a high yield.

  For example, in the case where the protease treatment is performed using an acidic protease and a neutral protease as described above, and then the treatment is performed using α-amylase, the pH is set to about 4. after the protease treatment. The amylase treatment may be performed by adjusting to 5 to 7.0.

  One amylase may be used alone, or multiple amylases may be used in combination as long as they do not adversely affect the action of amylases or proteases. In addition, amylase may be used in a free state or immobilized, and in any case, amylase is used in the range of about 0.1 to 5.0 g per 100 g of dried wheat protein. Good. The treatment with amylase can be terminated at any time. For example, when α-amylase is used, the treatment can be deactivated by setting the temperature of the solution to about 60 to 100 ° C.

(3) As a method for hydrolyzing wheat protein using an acid, a conventional method can be employed.
For example, as the acid, mineral acids such as sulfuric acid, hydrochloric acid, nitric acid, phosphoric acid and sulfurous acid; organic acids such as oxalic acid, citric acid, acetic acid and formic acid can be used.

As a procedure for hydrolyzing wheat protein using such an acid, an acid is usually added to an aqueous medium containing wheat protein so that the acid normality is in the range of 0.01 to 2 N. Examples include a procedure of hydrolyzing at a temperature of 50 to 100 ° C. for 10 minutes to 6 hours. When it is desired to stop the hydrolysis, it can be stopped by adding a base and neutralizing.

When hydrolyzing with an acid, it is necessary to adjust appropriately according to the kind and normality of the acid. Usually, the wheat protein concentration in the aqueous medium should be 1.0 to 80% by mass. .

  The wheat protein hydrolyzate used in the present invention is preferably a water-soluble component, and can be obtained in a dissolved state in an aqueous medium by performing the hydrolysis treatment of wheat protein as described above. However, it is desirable to further purify by removing the insoluble matter in the aqueous medium by an appropriate method such as centrifugation, filtration or decantation after hydrolysis.

  The fat accumulation inhibitor and lipolysis accelerator of the present invention may be a solution in which the water-soluble component, which is a hydrolyzate of wheat protein obtained by the above treatment, is dissolved in an aqueous medium, and is further diluted with an appropriate medium. The solution may be a solution, but these solutions are treated by an appropriate method such as evaporation, freeze-drying, hot-air drying, vacuum drying, spray drying, etc. to remove the solvent, or after being bound or supported on an appropriate carrier, It can also be obtained as a solid by removing the solvent by the method as described above.

  This fat accumulation inhibitor and lipolysis promoter is made from wheat and is safe and good in flavor, so it can be ingested as it is, and it can be easily taken for a long time. .

  The fat accumulation inhibitor and lipolysis promoter are usually administered in the range of 0.1 to 40 g per day for an adult as the dry weight of the wheat protein hydrolyzate. The general daily dose is 1 to 10 g. However, since the hydrolyzate of wheat protein is highly safe using wheat as a raw material, the dose can be further increased. The daily dose may be taken once, but may be taken in several divided doses.

  It should be noted that the fat accumulation inhibitor and lipolysis promoter of the present invention include other fat accumulation inhibitors, lipolysis promoters, in addition to the additives described below, as long as the original fat accumulation inhibitory action and lipolysis promoter action are not inhibited In addition, fat metabolism improving substances and the like may be blended alone or in combination.

  Examples of the dosage form of the fat accumulation inhibiting and lipolysis promoter of the present invention include tablets, capsules, granules, powders, syrups, dry syrups, liquids, suspensions, inhalants, and other oral preparations; suppositories, etc. Enteral preparations; topical skin preparations such as ointments, creams, gels and patches; injections and the like. Of these, oral preparations are preferred.

  Such dosage forms of fat accumulation inhibitor and lipolysis accelerator include conventional excipients, disintegrants, binders, lubricants, surfactants, alcohol, water, water-soluble polymers, sweeteners, taste masking agents. Additives such as agents and acidulants can be blended according to the dosage form, and can be produced according to conventional methods. Liquid preparations such as liquids and suspensions may be dissolved or suspended in water or other suitable medium immediately before taking, and tablets and granules may be coated on the surface by a well-known method. May be.

  When the fat accumulation-suppressing and lipolysis promoter of the present invention contains the additive and other fat accumulation-inhibiting substances, lipolysis promoting substances, fat metabolism improving substances, etc., the content of the wheat protein hydrolyzate is: Although it varies depending on the dosage form, it is usually in the range of 0.0001 to 99% by mass, preferably 0.1 to 80% by mass. It is desirable that the daily dose can be controlled so that the dose of wheat protein hydrolyzate (dry mass) can be taken.

  The fat accumulation inhibitor and lipolysis promoter of the present invention have an action of suppressing new accumulation of body fat and reducing body fat that has already accumulated, and therefore are suitably used as an obesity preventing agent or an obesity improving agent. be able to. The fat accumulation inhibitor and lipolysis promoter does not inhibit differentiation from preadipocytes into adipocytes, and therefore when used as an obesity preventive or obesity-improving agent, the fat cell functions as a secretory organ. There is an advantage that a slimming effect can be obtained by preventing or improving obesity healthily without damaging it.

  Furthermore, a method for preventing or improving obesity using the obesity-preventing agent or obesity-improving agent is provided by ingesting the obesity-preventing agent or obesity-improving agent in a form capable of managing the above-mentioned daily dose. Is done.

  The food and drink containing the fat accumulation inhibitor and lipolysis promoter of the present invention include health foods, functional foods, foods for specified health use, etc. that promote health through obesity-preventing action or obesity-improving action. All the food-drinks which can mix | blend accumulation | storage suppression and a lipolysis promoter are mentioned.

Specifically, tablets, tablet confectionery, chewable tablets, powders, capsules, granules, liquid foods such as tube enteral nutrients, health foods or dietary supplements such as drinks; green tea, oolong tea, black tea, etc. Tea drinks, soft drinks, jelly drinks, sports drinks, milk drinks, carbonated drinks, fruit juice drinks, lactic acid bacteria drinks, fermented milk drinks, powdered drinks, cocoa drinks, purified water drinks; butter, jam, sprinkles, margarine spreads Kinds: mayonnaise, shortening, custard cream, dressings, breads, cooked rice, noodles, pasta, miso soup, tofu, milk, yogurt, soup or sauces, confectionery (eg biscuits and cookies, chocolate, candy, cake, ice cream) Cream, chewing gum, tablet) and the like.

  The food / beverage products of the present invention, in addition to the above-mentioned fat accumulation suppression and lipolysis promoter, other food / beverage material materials used in the production of the food / beverage products, various nutrients, various vitamins, minerals, dietary fiber, various additives ( For example, it can be produced according to a conventional method by blending a taste ingredient, a sweetener, an acidulant such as an organic acid, a stabilizer, and a flavor).

  In this food and drink, the content of the wheat protein hydrolyzate varies depending on the form of the food and drink, but is usually in the range of 0.0001 to 90 mass%, preferably 0.1 to 70 mass%, and accumulates fat. It is desirable that the dosage per day can be controlled so that the dosage of the hydrolyzate (dry mass) of wheat protein per day described above as the dosage of the inhibitor and lipolysis promoter can be ingested.

By ingesting the food or drink in a form capable of managing the above-mentioned daily dose, a method for preventing or improving obesity using the food or drink is provided.
The present invention will be described with reference to the following examples, but the present invention is not limited to the following examples.

In the following examples, the content of the peptide mixture in the hydrolyzate of wheat protein and the molecular weight of the peptide mixture were measured by the following methods.
[Content of peptide mixture in hydrolyzate of wheat protein]
50 mg of wheat protein hydrolyzate was dissolved in 1 mL of distilled water to obtain a sample solution, and 1.5 mol / L of trichloroacetic acid equivalent to the sample solution was added to the sample solution and sufficiently stirred. Subsequently, this was centrifuged at 10,000 rpm for 5 minutes, the supernatant was collected, diluted 50 times with distilled water, and used as a sample for measurement.

The content of the peptide mixture is determined using the DC-Protein assay kit (manufactured by BIO-RAD).
Then, the absorbance at 750 nm was measured by adding a reagent provided to the sample and calculated from a calibration curve using albumin.

[Molecular weight measurement of peptide mixture contained in wheat protein hydrolyzate]
10 mg of wheat protein hydrolyzate is dissolved in 1 mL of distilled water, subjected to size exclusion high performance liquid chromatography (column used; Asahipak GS-320HQ (Shodex)), and then 0.1% trifluoroacetic acid solution / 40% acetonitrile. The aqueous solution was used to elute at a flow rate of 1.0 mL / min. Ultraviolet absorption at 220 nm of the eluted liquid was measured, and the measured absorbance was measured as Gly-Gly (molecular weight 132.12 Da), Gly-Gly-Gly (molecular weight 189.17 Da), Glutathione (molecular weight 612 Da) and Bacitracin. The molecular weight of the peptide mixture contained in the hydrolyzate of wheat protein was determined by applying a calibration curve prepared from (molecular weight 1422 Da).

[Production example]
(1) A reaction kettle is charged with 300 g of wheat gluten (active gluten, manufactured by Weston Foods Limited), 1920 mL of water, and 7.0 g of citric acid, and the pH is adjusted to 4.0 using citric acid. (Amano, Amanoenzyme) 0.45 g (20,000 units) was added, and the dispersion was heated to 45 ° C. and reacted for 5 hours.

(2) Next, the dispersion was adjusted to pH 7.5 using sodium hydroxide, and the temperature of the dispersion was kept at 85 ° C. for 10 minutes to inactivate the acidic protease.
(3) Next, the temperature of the dispersion is adjusted to 65 ° C., 0.45 g (320,000 units) of neutral protease (thermoase, Yamato Kasei) is added, and the mixture is reacted for 5 hours. The neutral protease was inactivated by maintaining at 20 ° C. for 20 minutes.

(4) Thereafter, the dispersion was centrifuged (8,000 G, 30 minutes) to remove insoluble matters, and the supernatant was recovered to obtain a water-soluble component.
(5) The obtained water-soluble component was freeze-dried to obtain about 166 g of a solid wheat protein hydrolyzate.

  (6) The content of the peptide mixture in the solid wheat protein hydrolyzate obtained in (5) above was measured and found to be about 80% by mass. Moreover, when the molecular weight of the peptide mixture contained in the hydrolyzate of the wheat protein was measured, the molecular weight was distributed to about 10,000 Da or less, and 76% by mass of the peptide mixture was within a molecular weight range of 200 to 2,000 Da. there were. Therefore, the solid wheat protein hydrolyzate obtained in (5) contained about 61% by mass (= 80 × 0.76) of a peptide mixture having a molecular weight of 200 to 2,000 Da.

[Example 1]
Manufacture of tablets 84 g of the hydrolyzate of wheat protein obtained in the production example, 10 g of crystalline cellulose (Asahi Kasei) and 5 g of polyvinyl pyrrolidone (BASF) were added, and 30 mL of ethanol was added thereto, followed by a wet method according to a conventional method. Granules were produced. After the granules were dried, 1.2 g of magnesium stearate was added to obtain granules for tableting, and tableting was performed using a tableting machine to produce 100 tablets each having 1 g.

[Example 2]
Production of granule 100 g of wheat protein hydrolyzate obtained in the same manner as in the production example, 100 g of lactose (DMV) and 40 g of crystalline cellulose (Asahi Kasei) are mixed, and 130 ml of ethanol is added to the kneader. The kneading was conducted for 5 minutes. After kneading, the mixture was sieved with 10 mesh and dried at 50 ° C. in a dryer. After drying, the particles were sized to obtain 240 g of granules.

[Example 3]
Manufacture of syrup agent While boiling 400 g of purified water and stirring it, add 750 g of sucrose and 100 g of the hydrolyzate of wheat protein obtained in the same manner as in the production example, dissolve it, and wipe it with heat. Purified water was added to make a total volume of 1000 mL to produce a syrup.

[Example 4]
Manufacture of liquid food In 700 g of pure water at about 65 ° C., 40 g of sodium caseinate (DMV), 160 g of maltodextrin (Sanwa starch) and 50 g of wheat protein hydrolyzate obtained in the production example were added and dissolved, and then vitamins were added. Mix and each component mixture of trace minerals were added. The obtained mixed solution was put into a homomixer and roughly emulsified at about 8,000 rpm for 15 minutes. The obtained emulsified liquid was cooled to about 20 ° C., and after adding a fragrance, the final volume was increased. After filling this solution with 230 g of this solution in a pouch, the pouch was sealed with nitrogen substitution, and sterilized at 121 ° C. for 15 minutes to obtain a liquid food.

[Example 5]
Production of bread 160 g of flour (strong flour) and 2 g of dry yeast were mixed. Separately, 20 g of the wheat protein hydrolyzate obtained in the production example, 25 g of sugar, 3 g of salt and 6 g of skim milk powder were dissolved in 70 g of hot water, and 1 egg was added and mixed well. This was added to the above mixture of wheat flour and dry yeast, kneaded by hand, and then kneaded with about 40 g of butter to prepare 20 roll bread dough. Next, these bread dough was fermented, and then a beaten egg was applied on the surface and baked in an oven at 180 ° C. for about 15 minutes to prepare a roll. Appearance, taste and texture were good.

[Example 6]
Manufacture of retort rice After adding 2g of wheat protein hydrolyzate obtained in the production example to rice and adding 2g of cooked rice, and filling this into a retort pack according to conventional methods The mixture was sealed with nitrogen substitution and sterilized at 121 ° C. for 15 minutes to obtain retort rice. The obtained cooked rice of retort rice had good appearance, taste and texture.

[Example 7]
Production of sauce for pasta A serving portion of meat sauce for pasta (150 g) was placed in a pan, and 5 g of the wheat protein hydrolyzate obtained in Production Example was added thereto and heated. After filling this pouch with a pouch, the pouch was sealed with nitrogen substitution and sterilized at 121 ° C. for 15 minutes to obtain a meat sauce for pasta.

[Test Example 1]
Cell differentiation inhibitory activity confirmation test (1) Differentiation and culture into adipocytes 3T3-L1 cells (mouse, fibroblast-like cells) were seeded in a 24-well plate, and differentiation induction medium (0.5 mM 1-methyl-3- The cells were cultured for 2 days in 10% FCS-added DMEM medium containing isobutylxanthine, 0.25 μM dexamethasone, 10 μg / mL insulin, and induced to differentiate into adipocytes. Thereafter, the medium was changed to a maturation promotion medium (10% FCS-added DMEM medium containing 5 μg / mL insulin) and cultured for 7 days.

  At this time, the wheat protein hydrolyzate obtained in the production example was added to the differentiation induction medium and the maturation promotion medium so as to be 1 mg / mL, and GPDH (glycerol-3 as a cell differentiation marker) after completion of the culture. -Phosphate dehydrogenase) activity was measured using a GPDH measurement kit (Hokudo Co., Ltd.). As a control group, one cultured in a medium to which no hydrolyzate of wheat protein was added was used.

(2) Measurement of GPDH activity After completion of the culture, the wells were washed twice with PBS (−), and then 0.5 mL of the enzyme extract was added, and the cells were detached from the well bottom by pipetting.

  Next, the peeled cells were put together with the enzyme extract into a sampling tube, and the sampling tube was immersed in liquid nitrogen for about 10 minutes to freeze the contents. Thereafter, the sampling tube was immersed in a water bath set at 37 ° C. to melt the contents. This freeze-thaw was repeated 3 times in total to disrupt the cells, and the resulting disrupted solution was centrifuged at 12,800 G, 4 ° C. for 5 minutes, and the supernatant was collected. The collected supernatant was used as a specimen.

  200 μL of the reaction substrate solution was placed in a 96-well plate and heated at 25 ° C. for about 5 minutes. Next, 200 μL of the specimen was placed in the 96-well plate and stirred well, and the decrease in absorbance at 340 nm was measured over time (10 minutes every other minute) to determine the amount of change in absorbance per minute. .

  The obtained change in absorbance was applied to the following formula to determine GPDH activity.

  The results are shown in Table 1 as relative values when the control group is 100%.

As is apparent from Table 1, 3T3-L1 cells cultured in a medium supplemented with wheat protein hydrolyzate showed 115% GPDH activity at 1 mg / mL compared to the control group. Therefore, it was clarified that the hydrolyzate of wheat protein has no cell differentiation-inhibiting activity from preadipocytes to adipocytes.

[Test Example 2]
Fat accumulation inhibitory action confirmation test (1) Differentiation and culture into adipocytes 3T3-L1 cells were seeded in a 24-well plate, and differentiation induction medium (0.5 mM 1-methyl-3-isobutylxanthine, 0.25 μM dexamethasone, 10 μg / mL
10% FCS-added DMEM medium containing insulin) was cultured for 2 days to induce differentiation into adipocytes. Thereafter, maturation promotion medium (DME supplemented with 10% FCS containing 5 μg / mL insulin)
M medium) and cultured for 7 days.

  At this time, the wheat protein hydrolyzate obtained in Production Example was added to the differentiation induction medium and the maturation promotion medium so as to be 0.01, 0.1, and 1 mg / mL. The amount of fat accumulated was measured. As a control group, one cultured in a medium to which no hydrolyzate of wheat protein was added was used.

(2) Preparation of reagent for measuring fat amount The amount of fat accumulated in fat cells was measured by staining intracellular lipid droplets with an oil red O staining solution and measuring the absorbance of the staining solution.

The oil red O staining solution stock solution is prepared by dissolving 0.5 g of oil red O staining solution powder in 100 mL of 2-propanol, and centrifuging at 3,000 rpm for about 5 minutes, and the supernatant with a syringe filter of 0.5 to 1.0 μm. Filtered and prepared. The oil red O staining solution was used by adding 4 mL of water to 6 mL of the stock solution at the time of measurement and allowing to stand for 1 hour, followed by filtration with a 0.5 to 1.0 μm syringe filter.

(3) Measurement of fat amount As described in (1) above, after cell culture in a differentiation induction medium and a maturation promotion medium for a total of 9 days, the culture solution was removed, and 500 μL of 10% formalin solution was added, and 4 The cells were fixed on the wells at 1 ° C for 1 hour. After removal of formalin, the plate was washed with distilled water, 300 μL of oil red O staining solution was added to the well, and left at room temperature for 15 minutes to stain intracellular fat. After removing the staining solution, the cells were washed with distilled water. Dye was extracted from the cells by adding 500 μL of 2-propanol to the wells, and the absorbance of the extract at 540 nm was measured with a plate reader.

  The results are shown in Table 2 as relative values when the control group is 100%.

As is clear from Table 2, the fat cells cultured in the medium supplemented with wheat protein hydrolyzate were 94% at 0.01 mg / mL, 87% at 0.1 mg / mL, 1 mg / mL as compared with the control group. mL
Only 83% of the fat accumulated, and it was revealed that the hydrolyzate of wheat protein exhibits a fat accumulation-inhibiting action.

[Test Example 3]
Lipolysis-promoting action confirmation test 3T3-L1 cells were seeded in a 24-well plate, and differentiation induction medium (0.5 mM 1-methyl-3-isobutylxanthine, 0.25 μM dexamethasone, 10 μg / mL)
10% FCS-added DMEM medium containing insulin) was cultured for 2 days to induce differentiation into adipocytes. Thereafter, maturation promotion medium (DME supplemented with 10% FCS containing 5 μg / mL insulin)
M medium) and cultured for 7 days.

  The medium was then removed and washed with PBS. 500 μL of 10% FCS-added DMEM medium containing 1 mg / mL wheat protein hydrolyzate obtained in Production Example was added to the wells and cultured for 6 hours. The control group was cultured in a medium containing no wheat protein hydrolyzate.

  Next, the medium was collected, and the amount of free glycerol in the medium degraded from adipocytes was measured using a glycerol measurement kit (F-kit glycerol, manufactured by J.K. International).

  The results are shown in Table 3 as relative values when the control group is 100%.

As is apparent from Table 3, adipocytes cultured in a medium containing a wheat protein hydrolyzate produced 113% free glycerol at 1 mg / mL compared to the control group. Therefore, it became clear that the hydrolyzate of the wheat protein exhibits lipolysis promoting activity.

From the above, fat accumulation suppression and lipolysis promoters, anti-obesity agents, obesity-improving agents, and foods and drinks containing a hydrolyzate of wheat protein inhibit the differentiation of preadipocytes into adipocytes. Thus, it has been clarified that there is an effect of suppressing the accumulation of fat in the fat cells and promoting the decomposition of the fat existing in the fat cells.

Claims (6)

  A fat accumulation inhibitor and a lipolysis promoter, characterized by comprising a hydrolyzate of wheat protein.   The fat accumulation inhibitor and lipolysis promoter according to claim 1, wherein the wheat protein hydrolyzate contains a peptide mixture having a molecular weight of 200 to 2,000 Da in an amount of 50 to 90% by mass.   3. The agent for inhibiting fat accumulation and promoting lipolysis according to claim 1 or 2, wherein the agent does not have a cell differentiation inhibitory activity from preadipocytes to adipocytes.   The fat accumulation inhibitor and lipolysis promoter according to any one of claims 1 to 3, which is an obesity preventive agent.   The fat accumulation inhibitor and lipolysis promoter according to any one of claims 1 to 3, which is an obesity-improving agent. A food or drink comprising the fat accumulation inhibitor and lipolysis promoter according to any one of claims 1 to 5.