JP2021119787A - Peptide composition and production method therefor - Google Patents

Abstract

To provide a peptide composition that includes a functional peptide.SOLUTION: The present invention provides a peptide composition that includes one or more components selected from the group consisting of: 0.00001-0.2 mass% of WY relative to the total peptide content; 0.000001-0.1 mass% of WL relative to the total peptide content; 0.000001-0.1 mass% of WV relative to the total peptide content; and 0.000005-0.2 mass% of GTWY relative to the total peptide content.SELECTED DRAWING: Figure 3

Description

The present invention relates to a peptide composition and a method for producing the same.

Maintenance, improvement, and improvement of brain function are required by a wide range of generations, from young people to the elderly. Maintaining, improving, and improving memory and learning abilities are important not only for students and working adults studying for entrance exams and qualification exams, but also for their daily work and life. In addition, since deterioration of memory and cognitive function are related to quality of life in the elderly, it is required to prevent, maintain, improve, and improve the deterioration of brain function.

Furthermore, in old age, an increase in mental disorders caused by a decrease in brain function due to aging has become a social problem as the number of elderly people increases. It has been reported that not only dementia represented by Alzheimer's disease but also mental illnesses such as depression and delirium are caused by the decrease in brain function as diseases caused by the decrease in brain function (non-patent documents). 1).

As the elucidation of brain function progresses, the search for substances that enhance brain function is being actively conducted. In particular, since there is no concern about side effects, analysis is being conducted to see if food ingredients contain substances that improve brain function. For example, there is described an attempt to increase the plasma tryptophan concentration by administering whey protein for the purpose of increasing the tryptophan concentration in the brain to induce an increase in serotonin concentration and affect the brain function (Non-Patent Document 2). ..

Patent Document 1 describes a composition capable of contributing to an increase in tryptophan concentration in the brain in which the value obtained by dividing the tryptophan content by the neutral large amino acid content is larger than a specific value, and Patent Document 2 is derived from lysozyme. It is described that the effect can be expected by ingesting one beverage containing several tens of mg of this composition per day. Patent Document 3 describes that a composition in which the value obtained by dividing the tryptophan content by the neutral large amino acid content is larger than a specific value can be obtained by hydrolyzing α-lactoglobulin or the like preferably with an acidic protease. There is.

Japanese Patent Publication No. 2011-523547 Special Table 2015-524265 Japanese Patent No. 4642321

Toshihisa Tanaka, Masatoshi Takeda (2011) Japanese Clinic, Special Issue, pp. 52-56 Am.J.Clin.Nutr., 2005,81,1026-33

Now, the present inventors have found that a peptide composition composed of an enzymatically decomposed product of whey protein and the peptide in the peptide composition are effective for maintaining, improving, improving and the like of brain function. The present inventors have also found a method for efficiently producing a peptide composition having functions such as maintenance, improvement and improvement of brain function. The present invention is based on these findings.

That is, an object of the present invention is to provide a peptide composition containing a functional peptide and a method for producing the same.

According to the present invention, the following inventions are provided.
[1] WY of 0.00001 to 0.2% by mass based on the total peptide content, WL of 0.000001 to 0.1% by mass based on the total peptide content, 0. One or more components selected from the group consisting of 0.000001 to 0.1% by weight of WV and 0.000005 to 0.2% by weight of GTWY (SEQ ID NO: 1) relative to total peptide content (preferably). Is a peptide composition containing the above four peptide components).
[2] KPTPEGDLEI (SEQ ID NO: 2) and / or GYGGVSLP (SEQ ID NO: 3) is further included, and the KPTPEGDLEI content is 0.000001 to 0.05% by mass based on the total peptide content, and the GYGGVSLP content. The peptide composition according to the above [1], wherein is 0.000005 to 0.2% by mass.
[3] The above-mentioned [1] or [2], wherein the total content of WY, WL, WV, GTWY, KPTPEGDLEI and GYGGVSLP is 0.00002 to 0.6% by mass with respect to the total peptide content. Peptide composition.
[4] The peptide composition according to any one of the above [1] to [3], which is in the form of a solution.
[5] In the solution, the WY concentration is 0.15 ppm or more, the WL concentration is 0.15 ppm or more, the WV concentration is 0.1 ppm or more, and the GTWY concentration is 0.3 ppm or more. The peptide composition according to the above [4].
[6] The peptide composition according to the above [4] or [5], wherein the GTWY concentration in the solution is 3 ppm or more.
[7] The peptide composition according to any one of [4] to [6] above, wherein the KPTPEGDLEI concentration in the solution is 0.1 ppm or more and the GYGGVSLP concentration is 0.1 ppm or more.
[8] The peptide composition according to any one of [4] to [7] above, wherein the total of WY concentration, WL concentration, WV concentration, GTWY concentration, KPTPEGDLEI concentration and GYGGVSLP concentration in the solution is 1 ppm or more.
[9] The peptide composition according to any one of [4] to [8] above, wherein the total of WY concentration, WL concentration, WV concentration and GTWY concentration in the solution is 0.5 ppm or more.
[10] The peptide composition according to any one of claims [4] to [9] above, wherein the total of the WY concentration, the WL concentration and the WV concentration in the solution is 0.5 ppm or more.
[11] One or more selected from the group consisting of 160 to 70,000 μg WY, 100 to 60,000 μg WV, 100 to 60,000 μg WL and 50 to 40,000 μg GTWY in the peptide composition in the ingestion form. A peptide composition comprising an ingredient.
[12] The peptide composition according to the above [11], wherein the content of GTWY is 100 to 30,000 μg.
[13] The peptide composition according to the above [11] or [12], further comprising GYGGVSLP and / or KPTPEGDLEI, having a GYGGVSLP content of 50,000 to 50,000 μg and a KPTPEGDLEI content of 160 to 70,000 μg. ..
[14] The peptide composition according to any one of the above [1] to [13], which is in the form of food.
[15] The composition according to the above [14], wherein the food is a beverage.
[16] A food product containing the peptide composition according to any one of the above [1] to [13].
[17] The food according to the above [16], which is a beverage.
[18] The peptide composition according to any one of the above [1] to [15] or the food according to the above [16] or [17] for maintaining, improving and / or improving brain function.
[19] An effective amount of the peptide composition according to any one of the above [1] to [15] or the food according to the above [16] or [17] is ingested or administered to a human or non-human animal. How to maintain brain function, how to improve brain function, and how to improve brain function.
[20] Use of the peptide composition according to any one of the above [1] to [15] for the production of compositions and foods for maintaining, improving and / or improving brain function.
[21] Any of the above [1] to [15], which comprises a step of reacting whey protein with an enzyme preparation containing one or more kinds of proteolytic enzymes derived from Aspergillus meleus to obtain an enzymatic degradation product. The method for producing a peptide composition according to the above.
[22] The production according to the above [21], wherein the step of allowing the enzyme to act on the whey protein is carried out by holding the whey protein at an action temperature of 45 to 55 ° C. for a certain period of time and then holding the whey protein at an action temperature of 60 to 75 ° C. for a certain period of time. Method.
[23] The production method according to the above [21] or [22], further comprising a step of diluting an enzymatic degradation product of whey protein or a step of concentrating an enzymatic degradation product of whey protein.

According to the present invention, there are 6 kinds of peptides capable of exerting functions such as maintenance, improvement and improvement of brain function, and a peptide composition containing a part or all of them. Since the six peptides of the present invention and the peptide composition of the present invention can utilize enzymatically decomposed products of foods, they are advantageous in that they can be used as functional materials that are safe for mammals including humans.

(A) is a figure which showed the outline of the Y-shaped maze test. (B) is a diagram showing a schematic diagram of a Y-shaped maze test. It is a figure which showed the volatility (A) of the spontaneous exchange behavior of the mouse which ingested a whey decomposition product, and the total number of invasion into an arm (B). It is a figure which showed the volatility of volatility exchange behavior (A) of the mouse which ingested GTWY, and the total number of invades into an arm (B). It is a figure which showed the volatility of volatility exchange behavior (A) of the mouse which ingested GYGGVSLP and the total number of invasion to the arm (B). It is a figure which showed the spontaneous exchange behavior volatility (A) and the total number of entry into an arm (B) of the mouse which ingested KPTPEGDLEI. It is a figure which showed the volatility (A) of the volatility exchange behavior of the mouse which ingested WY, WL and WV and the total number of invasion to the arm (B).

Specific description of the invention

According to the first aspect of the present invention, a peptide composition comprising one, two, three or four components selected from the group consisting of WY, WL, WV and GTWY as the peptide (hereinafter, "" (Sometimes referred to as the "first peptide composition") is provided. In this peptide composition, the WY content is 0.00001 to 0.2% by mass (preferably 0.00002 to 0.15% by mass, more preferably 0) with respect to the total peptide content in the composition. .05 to 0.2% by mass), and the WL content is 0.000001 to 0.1% by mass (preferably 0.000002 to 0.05% by mass, more preferably 0.025 to 0.055% by mass). %), The WV content is 0.000001 to 0.1% by mass (preferably 0.000002 to 0.05% by mass, more preferably 0.015 to 0.045% by mass), and the GTWY content is contained. The amount is 0.000005 to 0.2% by mass (preferably 0.00001 to 0.15% by mass).

Here, the "total peptide content" means the content of a peptide having a molecular weight of less than 10 kDa among the compounds having amino nitrogen. Further, as will be described later, since the first peptide composition of the present invention can be preferably derived from a whey decomposition product, the peptide that is the basis for calculating the "total peptide content" is derived from a whey decomposition product. Can be.

According to a preferred embodiment of the first peptide composition of the present invention, a peptide composition containing at least WY, WL, WV and GTWY as peptides, which contains WY content, WL content, WV content and GTWY. Peptide compositions are provided in amounts as described above.

The first peptide composition of the present invention can exhibit functions such as maintenance, improvement and improvement of brain function by ingesting it as it is or by concentrating or diluting it in mammals including humans. Therefore, the first peptide composition of the present invention can be used as a raw material for functional foods and pharmaceuticals, and can also be used as functional foods and pharmaceuticals.

The first peptide composition of the present invention may contain components other than peptides, but the total peptide content with respect to the total solid content mass of the composition can be 0.005% or more, preferably 0.01. % Or more, more preferably 0.05% or more. The upper limit can be 100%, and in this case, all the solids in the composition are composed of peptides. Here, the total peptide content can be measured by a conventional method such as amino acid analysis and ultraviolet absorbance measurement.

In the first peptide composition of the present invention, the total content of WY, WL, WV and GTWY with respect to the total peptide content can be 0.00005 to 0.4% by mass, preferably 0.00002 to 0.00002 to 0.4% by mass. It is 0.35% by mass, more preferably 0.2 to 0.35% by mass. In this case, the first peptide composition of the present invention is preferably a peptide composition containing at least WY, WL, WV and GTWY, and more preferably 0.000001 to 0.000001 to the total peptide content. 0.2% by mass WY, 0.000001-0.1% by mass WL with respect to total peptide content, 0.000001-0.05% by mass WV with respect to total peptide content and total peptide content It is a peptide composition containing 0.000005 to 0.2% by mass of GTWY.

The first peptide composition of the present invention may further comprise KPTPEGDLEI and / or GYGGVSLP in addition to WY, WL, WV and GTWY. The KPTPEGDLEI content relative to the total peptide content can be 0.000001 to 0.05% by weight (preferably 0.0000015 to 0.04% by weight), and the GYGGVSLP content relative to the total peptide content is 0.000001. It can be ~ 0.2% by mass (preferably 0.000005 to 0.1% by mass).

That is, according to a preferred embodiment of the first peptide composition of the present invention, it is a peptide composition containing at least WY, WL, WV, GTWY, KPTPEGDLEI and GYGGVSLP as peptides, and has a WY content and a WL content. , WV content, GTWY content, KPTPEGDLEI content and GYGGVSLP content are as described above.

In the first peptide composition of the present invention, the total content of WY, WL, WV, GTWY, KPTPEGDLEI and GYGGVSLP with respect to the total peptide content can be 0.00002 to 0.6% by mass, preferably 0.00002 to 0.6% by mass. It is 0.00004 to 0.5% by mass, more preferably 0.3 to 0.5% by mass. In this case, the first peptide composition of the present invention is preferably a peptide composition containing at least WY, WL, WV, GTWY, KPTPEGDLEI and GYGGVSLP, and more preferably with respect to the total peptide content. 0.00001-0.2% by weight WY, 0.000001-0.1% by weight WL with respect to total peptide content, 0.000001-0.05% by weight WV with respect to total peptide content, GTWY with a total peptide content of 0.000005 to 0.2% by weight, KPTPEGDLEI with a total peptide content of 0.000001 to 0.05% by weight, 0.000005 to 0 with respect to the total peptide content A peptide composition comprising .2% by weight of GYGGVSLP.

In the present invention, the content of WY, WL, WV, GTWY, KPTPEGDLEI and GYGGVSLP can be measured by liquid chromatography tandem mass spectrometry (LC / MS / MS). Those skilled in the art can easily set the conditions, but for example, the conditions of the examples described later can be used. At that time, it goes without saying that a standard peptide having a purity for LC / MS / MS measurement can be used, but for example, an AQUA peptide manufactured by SIGMA-ALDRICH can be used.

The first peptide composition of the present invention can be derived from an enzymatic degradation product of protein, preferably derived from an enzymatic degradation product of whey protein (sometimes referred to simply as "whey degradation product" in the present specification). be able to. The first peptide composition of the present invention contains any of the whey decomposition product itself, the concentrate of the whey decomposition product, and the dilution of the whey decomposition product. The method for producing a decomposition product of whey protein will be described later.

The first peptide composition of the present invention is not particularly limited in its form as long as the peptide is contained in a predetermined content ratio, and can be in the form of solid, semi-solid, powder or solution, for example. The solution is preferably an aqueous solution.

When the first peptide composition of the present invention is in the form of a solution, the concentration of the peptide in the composition can be as follows. That is, the concentration of WY can be 0.15 ppm or more (for example, 0.15 to 100 ppm), preferably 1.5 ppm or more (for example, 1.5 to 100 ppm), and more preferably 15 ppm or more (for example, 15). ~ 100 ppm). The concentration of WL can be 0.15 ppm or more (for example, 0.15 to 40 ppm), preferably 1.5 ppm or more (for example, 1.5 to 40 ppm), and more preferably 15 ppm or more (for example, 15 to 40 ppm). Is. The concentration of WV can be 0.1 ppm or more (for example, 0.1 to 30 ppm), preferably 0.5 ppm or more (for example, 0.5 to 30 ppm), and more preferably 1 ppm or more (for example, 1 to 30 ppm). Is. The concentration of GTWY can be 0.3 ppm or more (for example, 0.3 to 150 ppm), preferably 3 ppm or more (for example, 3 to 150 ppm), and more preferably 30 ppm or more (for example, 30 to 150 ppm).

When the first peptide composition of the present invention is in the form of a solution, it can contain KPTPEGDLEI and / or GYGGVSLP, and the concentration of KPTPEGDLEI in the composition is 0.1 ppm or more (for example, 0.1 to 30 ppm). It is preferably 1.0 ppm or more (for example, 1.0 to 30 ppm), more preferably 10 ppm or more (for example, 10 to 30 ppm), and the GYGGVSLP concentration is 0.1 ppm or more (for example, 0.1 to 100 ppm). ), It is preferably 1.0 ppm or more (for example, 1.0 to 100 ppm), and more preferably 10 ppm or more (for example, 10 to 100 ppm).

When the first peptide composition of the present invention is in the form of a solution, the total of WY concentration, WL concentration, WV concentration, GTWY concentration, KPTPEGDLEI concentration and GYGGVSLP concentration in the composition is 1.0 ppm or more (for example, 1). It can be 0 to 500 ppm), preferably 2 ppm or more (for example, 2 to 500 ppm), more preferably 10 ppm or more (for example, 10 to 500 ppm), and further preferably 100 ppm or more (for example, 100 to 500 ppm). .. In this case, the first peptide composition of the present invention is preferably a peptide composition in solution form comprising at least WY, WL, WV, GTWY, KPTPEGDLEI and GYGGVSLP, more preferably 0.15 ppm. A peptide composition in solution form comprising the above WY, 0.15 ppm or more WL, 0.1 ppm or more WV, 0.3 ppm or more GTWY, 0.1 ppm or more KPTPEGDLEI, and 0.1 ppm or more GYGGVSLP. be.

When the first peptide composition of the present invention is in the form of a solution, the total of WY concentration, WL concentration, WV concentration, and GTWY concentration in the composition is 0.5 ppm or more (for example, 0.5 to 300 ppm). It can be preferably 2 ppm or more (for example, 2 to 300 ppm), more preferably 10 ppm or more (for example, 10 to 300 ppm), still more preferably 20 ppm or more (for example, 20 to 300 ppm), and most preferably 100 ppm or more (for example, 100 ppm or more). For example, 100 to 300 ppm). In this case, the first peptide composition of the present invention is preferably a peptide composition in solution form containing at least WY, WL, WV and GTWY, and more preferably 0.15 ppm or more of WY. A peptide composition in solution form comprising 0.15 ppm or more of WL, 0.1 ppm or more of WV and 0.3 ppm or more of GTWY.

When the first peptide composition of the present invention is in the form of a solution, the total WY concentration, WL concentration and WV concentration in the composition may be 0.5 ppm or more (for example, 0.5 to 200 ppm). It can be preferably 2 ppm or more (for example, 2 to 200 ppm), more preferably 5 ppm or more (for example, 5 to 200 ppm), further preferably 50 ppm or more (for example, 50 to 200 ppm), and most preferably 100 ppm or more (for example, 100 to 100 ppm). 200 ppm). In this case, the first peptide composition of the present invention is preferably a peptide composition in solution form containing at least WY, WL and WV, more preferably 0.15 ppm or more of WY, 0. A peptide composition in solution form comprising 15 ppm or more of WL and 0.1 ppm or more of WV.

The method for producing the first peptide composition of the present invention is not particularly limited as long as it contains a predetermined peptide in a predetermined content ratio, but the first peptide is formed by allowing an enzyme preparation containing a proteolytic enzyme to act on the raw material protein. The composition can be produced.

The enzyme treatment of the raw material protein can be carried out as follows. First, as the raw material protein, whole milk, milk powder, casein or whey can be used, and whey is preferable. Here, "whey" is also referred to as whey, milky lotion or whey, and means an aqueous solution obtained by removing milk fat, casein and the like from milk. Whey is composed of β-lactoglobulin, α-lactalbumin, serum albumin, immunoglobulin and the like. The animals and plants from which the whey used in the present invention is derived may be used, but it is preferable to use whey derived from milk.

When whey protein is used as a raw material, the concentration of whey protein subjected to the enzymatic reaction is not limited as long as the protein can be dissolved, but from the viewpoint of suppressing gelation and aggregation and saving the trouble of concentration, 1 to 30 w / It is preferably v%, more preferably 1 to 20 w / v%, and even more preferably 5 to 15 w / v%.

The raw material protein may be dissolved in any aqueous solvent as long as the enzymatic reaction proceeds, but it is preferably dissolved in water or a food additive grade buffer solution in consideration of its use as a food. It is preferable to use a buffer solution so that the pH of the reaction solution does not change due to the amino acids generated in the enzymatic reaction. The type of buffer solution is arbitrary and may be determined in consideration of subsequent use and the amount of flavor, taste and minerals, but the pH of the reaction solution is 4 to 9, preferably 5 to 8, more preferably 7 to 8. The composition is preferable so that it can be maintained at. Most preferably, it is a potassium phosphate buffer solution. The concentration of the buffer solution is arbitrary as long as the buffering effect can be obtained, but it can be 0.01 to 0.5 M, preferably 0.05 to 0, in consideration of the flavor, taste, and amount of minerals. It is 2M, more preferably about 0.1M.

The enzyme can be any enzyme preparation containing a proteolytic enzyme, but an enzyme preparation containing a neutral protease is preferable. As the enzyme preparation, for example, those derived from microorganisms such as Bacillus subtilis, Aspergillus oryzae, and Alpergillus meleus can be used. Among them, the enzyme preparation derived from Aspergillus oryzae and the enzyme preparation derived from Alpergillus meleus are used. Preferably, more preferably, it is an enzyme preparation derived from Alpergillus meleus.

In the present invention, a commercially available enzyme preparation can be used, and for example, the enzyme preparation can be obtained from Amano Enzyme, Shin Nihon Kagaku Kogyo, DSM, Danisco, Novozyme, HBI, and the like. The amount of the enzyme preparation added is arbitrary, but considering an appropriate hydrolysis reaction rate and cost, for example, 0.01 to 5 w / v%, preferably 0.05 to 4 w / v%, more preferably 0. It can be 1 to 0.5 w / v%.

The enzyme reaction temperature and the enzyme reaction time can be set so that the raw material protein is sufficiently hydrolyzed and the quality of the enzymatic decomposition product is maintained. That is, the enzyme reaction temperature can be, for example, 30 to 70 ° C, preferably 40 to 70 ° C, and more preferably 45 to 65 ° C. The enzyme reaction time can be 1 to 12 hours, preferably 2 to 10 hours, and more preferably 4 to 5 hours.

The enzymatic reaction is preferably carried out while raising the temperature. For example, the reaction can be carried out while raising the temperature from 30 ° C. to 75 ° C. over 4 to 10 hours. Preferably, the reaction can be carried out while raising the temperature from 35 ° C. to 75 ° C. for 5 to 8 hours, and more preferably, the reaction can be carried out from 35 ° C. to 75 ° C. for 6 to 8 hours. The temperature rise speed is arbitrary, but the holding time between 45 ° C and 55 ° C is lengthened (5 to 7 hours), then the temperature is raised quickly to 60 ° C, and then the temperature rise is lengthened between 60 ° C and 75 ° C. It is preferable to hold (for example, 1 to 3 hours). The most preferable method is to add the enzyme at 50 ° C., hold it for 5 to 7 hours, raise the temperature at an arbitrary rate, and hold it at a target temperature of 60 to 65 ° C. or 65 to 75 ° C. for 1 to 3 hours.

In the reaction, it is preferable to stir the reaction solution from the viewpoint of reaction efficiency. The liquid stirring speed should be high so that the substrate is in good contact with the enzyme, but if it is too fast, the reaction liquid may scatter. Therefore, for example, it can be set to 100 to 500 rpm, preferably 200 to 400 rpm. More preferably, it is about 250 rpm.

Once the desired peptide is obtained, the reaction solution is preferably subjected to an enzymatic reaction in a stop step. In the enzyme reaction termination step, a method of raising the temperature of the reaction solution, adding a chelating agent to change the chemical structure of the enzyme, or removing the enzyme by membrane treatment can be adopted. The preferred method is deactivation treatment at high temperature. The method can be carried out by holding at 80-90 ° C. for 5-30 minutes, preferably at 80-90 ° C. for 20-30 minutes. Further, when the temperature becomes high in the concentration step described later, the concentration step can also be performed.

The reaction solution (peptide composition) that has undergone the enzyme reaction step and the enzyme reaction termination step may be further subjected to a sterilization step. Examples of the sterilization step include a membrane treatment step and a high-temperature sterilization step described later. The heat sterilization step can also serve as an enzyme reaction termination step, which is advantageous in terms of simplification of the manufacturing process.

The reaction solution (peptide composition) that has undergone the enzyme reaction step and the enzyme reaction termination step may be further subjected to a purification step. Examples of the purification step include a membrane treatment step, and a preferable membrane treatment is ultrafiltration. The fractional molecular weight of the ultrafiltration is preferably 3 to 100 kDa, more preferably 5 to 50 kDa, and most preferably 10 kDa. Carrying out the purification step is advantageous in that the flavor of the peptide composition can be improved as compared to the case where it is not carried out. In addition, the purification step can also serve as an enzyme reaction termination step and a sterilization step, which is advantageous in terms of simplification of the manufacturing step.

The reaction solution (peptide composition) that has undergone the enzyme reaction step and the enzyme reaction termination step may be further subjected to a concentration step from the viewpoint of storage and transportation. Any method can be selected for the concentration step, but a method of concentration under reduced pressure, freeze-drying and spray drying (spray drying), and concentration by membrane treatment (for example, a method using a reverse osmosis membrane) is preferable, and more preferably freezing. Drying and spray drying. Spray drying is particularly preferable from the viewpoint of performing concentration in a large amount and efficiently. Further, the reaction solution (peptide composition) that has undergone the enzyme reaction step and the enzyme reaction termination step may be diluted with a solvent to be provided as a food or drink (particularly a beverage). The solvent used is not limited, but an aqueous solvent is preferable, and examples thereof include water, a sugar-containing aqueous solution, a sugar- and acidity-contributing substance-containing aqueous solution, and an arbitrary drinking liquid.

According to a preferred embodiment of the first aspect of the present invention, whey protein (more preferably Fontera WPC472) is dissolved in 0.1 M potassium phosphate buffer (pH 7.0) to a concentration of 10% and Amano. The present invention is obtained by adding Enzyme Protease P "Amano" 3SD to 0.2%, holding at 50 ° C. for 5 hours while stirring at 250 rpm, and then raising the temperature and holding at 60 ° C. for 3 hours. The first peptide composition of the above can be produced.

According to the second aspect of the present invention, an ingestion form comprising a specific amount of 1, 2, 3 or 4 peptides selected from the group consisting of WY, WL, WV and GTWY as peptides. Peptide composition (hereinafter sometimes referred to as "second peptide composition") is provided. In the second peptide composition, the content of the peptide in the composition can be set as follows.
WY: 160 to 70,000 μg (preferably 4000 to 60000 μg)
WV: 100 to 60,000 μg (preferably 160 to 55,000 μg)
WL: 100 to 60,000 μg (preferably 160 to 55,000 μg)
GTWY: 50-40,000 μg (preferably 100-30000 μg)

The second peptide composition of the present invention may further comprise GYGGVSLP and / or KPTPEGDLEI in addition to one or more peptides selected from the group consisting of WY, WV, WL and GTWY. The content of GYGGVSLP can be 50,000 to 50,000 μg (preferably 160 to 40,000 μg), and the content of KPTPEGDLEI can be 160 to 70,000 μg (preferably 270 to 60,000 μg).

The method for producing the second peptide composition of the present invention is not particularly limited as long as it contains a predetermined peptide in a predetermined content, but can be produced in the same manner as the first peptide composition. That is, an enzyme preparation containing a proteolytic enzyme is allowed to act on a raw material protein such as whole milk, milk powder, casein or whey, and in some cases, it is concentrated or diluted so that a predetermined peptide is contained in a predetermined content. Can be manufactured. For details of the enzymatic reaction, the description of the method for producing the first peptide composition can be referred to.

Since the second peptide composition of the present invention is ingested, it can be ingested as it is by mammals including humans, and can exhibit functions such as maintenance, improvement and improvement of brain function. Therefore, the second peptide composition of the present invention can be used as a functional food or a pharmaceutical product.

As shown in Examples below, enzymatic degradation products of whey protein and WY, WL, WV, GTWY, KPTPEGDLEI and GYGGVSLP (hereinafter sometimes referred to as "six peptides of the present invention") maintain and improve brain function. And has an improving effect. Therefore, the first peptide composition and the second peptide composition of the present invention and some or all of the six peptides of the present invention are active ingredients of the composition for maintaining, improving and / or improving brain function. It can be used as a method for maintaining brain function, a method for improving brain function, and a method for improving brain function. In addition, a part or all of the first peptide composition, the second peptide composition, and the six peptides of the present invention are used as active ingredients of a brain function maintaining agent, a brain function improving agent, and a brain function improving agent. Can be used. In the present invention, "brain function" is used in the sense of including memory learning function and cognitive function.

That is, according to the present invention, a human or non-human animal is allowed to ingest an effective amount of the first peptide composition, the second peptide composition of the present invention, or a part or all of the six peptides of the present invention. Alternatively, a method for maintaining brain function, a method for improving brain function, and a method for improving brain function are provided, which comprises administering. When these methods of the present invention are carried out, they can be carried out by administering an effective amount of the composition of the present invention or the peptide of the present invention to mammals including humans in need thereof.

According to the present invention, the first peptide composition of the present invention, the second peptide composition or the present invention for the production of compositions and foods for maintaining, improving and / or improving brain function. Use of some or all of the six peptides is provided. According to the present invention, the first peptide composition of the present invention, the second peptide composition, or the present invention for producing a brain function maintaining agent, a brain function improving agent, and a brain function improving agent. Use of some or all of the six peptides is provided. According to the present invention, further, the first peptide composition of the present invention, the second peptide composition and a part of the six peptides of the present invention for use in maintaining, improving and / or improving brain function or Everything is provided.

In the present invention, "maintenance of memory learning function" includes, for example, suppressing deterioration of memory ability and learning ability caused by aging in the elderly and the like. Further, "improvement of memory learning function" includes, for example, transient improvement of memory learning function, promotion of medium- to long-term memory retention, and promotion of brain development. Further, "improvement of memory learning function" includes, for example, recovery of a function that has once deteriorated due to aging or other factors or a function that has a sign of deterioration.

In the present invention, "maintenance of cognitive function" includes, for example, prevention of intellectual functions such as understanding, judgment, and logic, that is, deterioration of cognitive ability. In addition, "improvement of cognitive function" includes, for example, enhancing cognitive ability from the current level. Further, "improvement of cognitive function" includes, for example, recovery of a symptom that has once deteriorated cognitive ability or a sign of deterioration.

Without being bound by the following theory, it is included in these compositions that the first peptide composition and the second peptide composition of the present invention have a maintenance, improvement and ameliorating action on brain function. As a result of the peptide reinforcing the mechanism of the brain by promoting the production of neurotransmitters, it is considered that the peptide has led to the maintenance, improvement and improvement of brain function.

The use of the first peptide composition and the second peptide composition of the present invention and the six peptides of the present invention in the present invention may be used in humans and non-human animals, and may be used therapeutically and non-therapeutically. Both are intended for use. As used herein, "non-therapeutic" means does not include the act of surgery, treatment or diagnosis of a human (ie, medical practice for a human), specifically under the direction of a physician or physician. It means that a person does not include a method of performing surgery, treatment or diagnosis on a human.

In the present invention, improvement of brain function of a part or all of the first peptide composition and the second peptide composition of the present invention and the six kinds of peptides of the present invention is effective for the treatment, prevention or improvement thereof. A method of treating a disease or symptom that can be used as an active ingredient of a composition for treating, preventing and / or ameliorating a disease or symptom, and for which improvement of brain function is effective in treating, preventing or ameliorating the disease or symptom. , Can be used for preventive and ameliorating methods. In addition, the first peptide composition, the second peptide composition, and some or all of the six peptides of the present invention are therapeutic agents for diseases and symptoms for which improvement of brain function is effective for the treatment, prevention or improvement thereof. , Can be used as an active ingredient in prophylactics and improvers.

That is, according to the present invention, a human or non-human animal is allowed to ingest an effective amount of the first peptide composition, the second peptide composition of the present invention, or a part or all of the six peptides of the present invention. Provided are methods, preventive and ameliorating methods of treating, preventing or ameliorating diseases and conditions for which improvement of brain function is effective in treating, preventing or ameliorating brain function, including or including administration. When the therapeutic method, preventive method and improvement method of the present invention are carried out, it is carried out by administering an effective amount of the composition of the present invention or the peptide of the present invention to mammals including humans in need of treatment, prevention or improvement. can do.

According to the present invention, also according to the present invention, there is a invention for producing a therapeutic composition, a preventive composition and an improving composition of a disease and a condition for which improvement of brain function is effective for the treatment, prevention or improvement thereof. The use of one peptide composition, a second peptide composition or some or all of the six peptides of the invention is provided. These compositions are preferably pharmaceutical compositions. According to the present invention, the first peptide composition of the present invention for producing a therapeutic agent, a prophylactic agent and an ameliorating agent for a disease and a symptom for which improvement of brain function is effective for the treatment, prevention or amelioration thereof. The use of a second peptide composition or some or all of the six peptides of the invention is provided. According to the present invention, further, the first peptide composition of the present invention, the second peptide for use in the treatment, prevention and amelioration of diseases and symptoms for which improvement of brain function is effective for the treatment, prevention or amelioration thereof. The compositions and some or all of the six peptides of the invention are provided.

Diseases and symptoms for which improvement of brain function is effective for its treatment, prevention or improvement include diseases and symptoms for which improvement of memory learning function and cognitive function is effective for its treatment, prevention or improvement, and more specifically. Memory disorders (symptoms of being unable to remember or remember new things), aphasia (time / place / person's agnosia), cognitive dysfunction (decrease in computing ability / judgment) Aphasia, agnosia, apraxia, executive dysfunction).

The composition for maintaining, improving and / or improving the brain function of the present invention and the maintaining agent, improving agent and / or improving agent of the brain function of the present invention, and the improvement of the brain function of the present invention are the treatment, prevention or improvement thereof. Therapeutic compositions, prophylactic compositions and ameliorating compositions for diseases and symptoms that are effective in treating, preventing or ameliorating the improvement of brain function of the present invention. And the improving agent can be provided in the form of pharmaceuticals, non-pharmaceutical products, foods and drinks, feeds, etc., and can be carried out in accordance with the following description. Further, the method for maintaining the brain function, the method for improving the brain function, the method for improving the brain function, and the treatment method and the prevention method for the present invention can be carried out according to the following description.

The first peptide composition and the second peptide composition of the present invention, which are the active ingredients of the present invention, and some or all of the six peptides of the present invention are orally or parenterally administered to humans and non-human animals. It can be administered orally, preferably. Examples of oral preparations include granules, powders, tablets (including sugar-coated tablets), pills, capsules, syrups, emulsions, and suspensions. Parenteral preparations include injections (eg, subcutaneous injections, intravenous injections, intramuscular injections, intraperitoneal injections), infusions, and external preparations (eg, nasal preparations, transdermal preparations, ointments). ), Suppositories (eg, rectal suppositories, vaginal suppositories). These formulations can be formulated using a pharmaceutically acceptable carrier by a method usually used in the art. Pharmaceutically acceptable carriers include excipients, binders, diluents, additives, fragrances, buffers, thickeners, colorants, stabilizers, emulsifiers, dispersants, suspending agents, preservatives, etc. For example, magnesium carbonate, magnesium stearate, talc, sugar, lactose, pectin, dextrin, starch, gelatin, tragant, methyl cellulose, sodium carboxymethyl cellulose, low melting point wax, and cacao butter can be used as carriers.

The method for producing the oral preparation is not particularly limited, and any method can be used. Active ingredients include excipients (eg lactose, sucrose, starch, mannitol), disintegrants (eg calcium carbonate, carboxymethyl cellulose calcium), binders (eg pregelatinized starch, gum arabic, carboxymethyl cellulose, polyvinylpyrrolidone, etc. Hydroxypropyl cellulose), stabilizers and / or pharmaceutical additives such as lubricants (eg, starch, magnesium stearate, polyethylene glycol 6000) are compounded and compression molded, then taste masking, intestines, if necessary. It can be produced by coating by a known method for the purpose of solubility or persistence. As the coating agent, for example, ethyl cellulose, hydroxymethyl cellulose, polyoxyethylene glycol, cellulose acetate phthalate, hydroxypropyl methyl cellulose phthalate, Eudragit (Rohm Co., Ltd.) and the like can be used. Capsules may be produced by adding the active ingredient as it is or by blending the active ingredient with a pharmaceutical additive such as a stabilizer, a dispersant, a colorant and / or a preservative. Further, the capsule may be a soft capsule or a hard capsule, and the material of the capsule is not particularly limited, and for example, gelatin, vegetable fiber, starch or the like can be used, and adhesion prevention measures may be taken.

Of the parenteral agents, the injectable agent contains the active ingredient as a dispersant (for example, Tween80 (Atlas Powder)), a preservative (for example, methylparaben, propylparaben, benzyl alcohol, chlorobutanol, phenol), and an isotonic agent (for example). , Sodium chloride, glycerin, sorbitol, glucose, converted sugar, etc., along with aqueous solvents (eg distilled water, physiological saline, Ringer's solution) or oily solvents (eg olive oil, sesame oil, cottonseed oil, corn oil, etc.) It can be produced by dissolving, suspending or emulsifying in vegetable oil, propylene glycol, etc. At this time, if desired, additives such as a solubilizing agent (for example, sodium salicylate and sodium acetate), a stabilizer (for example, human serum albumin), and a soothing agent (for example, benzalkonium chloride and procaine hydrochloride) are added. May be good.

When a part or all of the first peptide composition and the second peptide composition which are the active ingredients of the present invention and the six kinds of peptides of the present invention are provided as foods, these shall be provided as they are as foods. Or these can be contained in foods and provided. The food provided in this manner is a food containing an effective amount of the first peptide composition, the second peptide composition, and a part or all of the six peptides of the present invention. As used herein, the term "containing an effective amount" of a part or all of the first peptide composition, the second peptide composition, and the six peptides of the present invention means an amount normally eaten in individual foods. The content is such that a part or all of the first peptide composition, the second peptide composition, and the six kinds of peptides of the present invention are ingested in the range as described later. In addition, "food" means health foods, functional foods, nutritional supplements, supplements, health functional foods (for example, specified health foods, nutritional functional foods, functional foods), special purpose foods (for example, infant foods). , Food for pregnant women, food for the sick).

Since some or all of the first peptide composition and the second peptide composition and some or all of the six peptides of the present invention have a function of maintaining, improving and / or improving brain function, daily foods and supplements are taken. It can be provided by containing it in a food to be ingested as. In this case, the first peptide composition, the second peptide composition, and some or all of the six peptides of the present invention may be provided in a predetermined unit packaging form in an amount to be ingested per meal. can. Examples of the unit packaging form per meal include a form in which a fixed amount is specified for packs, packages, cans, bottles, and the like. In order to better exert the various actions of the first peptide composition, the second peptide composition, and the six peptides of the present invention, the amount of the six peptides of the present invention to be ingested at one time will be described later. The intake per meal can be determined according to the above. The food product of the present invention may be provided with an explanation regarding the amount of intake displayed on the package or together with a document or the like containing the explanation.

The food product of the present invention may be labeled as having a maintenance, improvement and / or ameliorating effect on brain function. In this case, the food product of the present invention may be labeled in part or in whole as follows in order to make the labeling easy for consumers to understand. Needless to say, in the present invention, "maintenance, improvement and / or improvement of brain function" is used to include the following indications.
・ Improve memory ・ Enhance memory ・ Support memory ・ Improve cognitive function ・ Enhance cognitive function ・ Support cognitive function ・ Help maintain memory ・ Prevent inadvertentness ・ Improve memory retention Suppress the decline in memory associated with aging

The form of the "food" is not particularly limited, and may be, for example, a beverage form, a semi-liquid or a gel form. For example, a dairy product obtained by reacting a dairy material such as milk, yogurt, or cheese with the above-mentioned enzyme preparation to produce a part or all of the six peptides of the present invention is also within the scope of the food of the present invention. As a supplement, it is encapsulated in tablets or capsules produced by adding excipients, binders, etc. to a dry powder of some or all of the six peptides of the present invention, kneading them, and then tableting them. Examples thereof include capsules that have been prepared.

Further, it goes without saying that the food of the present invention can contain additives that are permissible for foods used in the production of ordinary foods, but these are from the viewpoint of preventing deterioration and oxidation of the peptide composition of the present invention. Ingredients expected to have the above effects may be added to the food product of the present invention. Examples of such a component include a flavor preparation having a peptide deterioration-preventing action and an antioxidant action, and a composition derived from a natural product can be preferably used.

The foods provided in the present invention include carbohydrate-containing foods and drinks such as rice, noodles, breads and pastas; Western confectionery such as cookies and cakes, Japanese confectionery such as buns and sheep syrup, candy, gums, yogurt and the like. Cold confectionery such as pudding and various confectioneries such as ice confectionery; whiskey, bourbon, spirits, liqueur, wine, fruit liquor, Japanese liquor, Chinese liquor, shochu, beer, non-alcoholic beer with alcohol content of 1% or less, sparkling liquor, and other miscellaneous liquors , Alcoholic beverages such as liquor high; beverages with fruit juice, beverages with vegetable juice, beverages with fruit juice and vegetable juice, soft drinks, carbonated beverages, milk, soy milk, milk beverages, drink-type yogurt, drink-type jelly, coffee, Non-alcoholic beverages such as cocoa, tea beverages, nutritional drinks, sports drinks, mineral water, near water; processed products using eggs, processed seafood and livestock meat (including guts such as liver) (including delicacies) Etc. can be exemplified, but the present invention is not limited to these. The mineral water includes both effervescent and non-effervescent mineral water.

The tea beverages include fermented teas, semi-fermented teas and non-fermented teas, for example, tea, green tea, barley tea, brown rice tea, roasted tea, tamaro tea, roasted tea, oolong tea, corn tea, puer tea, and louis bostee. , Rose tea, Kiku tea, herb tea (eg, mint tea, jasmine tea).

Fruits used in beverages containing fruit juice and beverages containing fruit juice and vegetable juice include, for example, apples, oranges, grapes, bananas, pears, peaches, mangoes, acai, blueberries and ume. Examples of vegetables used in beverages containing vegetable juice and beverages containing fruit juice and vegetable juice include tomatoes, carrots, celery, pumpkins, cucumbers and watermelons.

Since the drug and food of the present invention utilize an enzymatic decomposition product derived from milk that has been ingested by humans for many years as a food, the toxicity is low and the mammals (for example, human, mouse, rat, rabbit, dog) that require it are low in toxicity. , Cats, cows, horses, pigs, monkeys, etc.) can be safely used. The intake or dose of the six peptides of the present invention can be determined depending on the sex, age and body weight of the recipient, symptoms, administration time, dosage form, administration route, drugs to be combined, and the like. Examples of the intake and dose of the six peptides of the present invention per adult (50 kg) are as follows, but depending on the subject, they may be ingested or administered in several divided doses.
WY: 160-4200 μg (preferably 500-3600 μg)
WV: 100 to 3600 μg (preferably 160 to 3300 μg)
WL: 100 to 3600 μg (preferably 160 to 3300 μg)
GTWY: 50 to 2400 μg (preferably 100 to 1800 μg)
GYGGVSLP: 50-3000 μg (preferably 160-2400 μg)
KPTPEGDLEI: 160-4200 μg (preferably 270-3600 μg)

Since the first peptide composition and the second peptide composition of the present invention contain a part or all of the six kinds of peptides of the present invention, according to the dose and dose of each peptide at one time. The composition and intake or dose of the first peptide composition and the second peptide composition can be determined. For example, a first peptide composition in a unit-intake form or a unit-administration form or a second peptide in a unit-intake form or a unit-administration form so as to have a single intake and dose of each peptide as described above. The composition can be prepared.

Example 1: Examination of conditions for producing whey degradation products (1) Selection of raw material proteins Various whey proteins are enzymatically treated with an enzyme preparation containing a proteolytic enzyme (Protease P "Amano" 3SD or Samoaase C100 manufactured by Amano Enzyme). Then, the brain function active peptide produced was examined.

Specifically, the final concentration of each proteolytic enzyme preparation is 0.125 w / v% in a solution prepared by dissolving each whey protein in 0.05 M potassium phosphate buffer so as to be 5 w / v%. The enzyme preparation was added as described above. The obtained solution was reacted at pH 7.0 and 50 ° C. for 4 hours and then held at 90 ° C. for 20 minutes to inactivate the enzyme to obtain an enzyme-treated sample. The sample obtained by the same treatment except that the enzyme preparation was added was used as a control sample.

The results of visual observation of the obtained sample are shown in Table 1 below.

Further, the obtained sample was centrifuged, the supernatant was ultrafiltered (10 kDa), the obtained analytical sample was appropriately diluted, and the peptide was quantified by LC / MS / MS under the following analytical conditions. .. The concentration conversion was performed by the calibration curve method with the minimum concentration of 0.001 ppm.

From the results in Tables 1 and 2, some whey proteins (separated whey protein WP18855) aggregated due to heat treatment such as enzymatic reaction or enzyme inactivation (Table 1), but precipitation occurred in other areas. Peptide was also obtained, and the yield was not significantly different due to the difference in whey protein (peptide yield data omitted).

Therefore, it was found that almost all whey proteins can be used as raw materials in the production of the whey decomposition product of the present invention. Among them, it was found that WPC472 manufactured by Fonterra, which had less precipitation than the two enzyme preparations examined, was comprehensively suitable.

(2) Examination of enzyme preparation In 0.05M potassium phosphate buffer containing 5w / v% of whey protein (WPC472 manufactured by Fontera), the final concentration of proteolytic enzyme preparation is 0 for various enzyme preparations shown in Table 3. It was added so as to have a concentration of .125 w / v%. After reacting at pH 7.0 and 50 ° C. for 4 hours, the mixture was held at 90 ° C. for 20 minutes to inactivate the enzyme to obtain an enzyme-treated sample. The sample obtained by the same treatment except that the enzyme preparation was added was used as a control sample.

The obtained sample was ultrafiltered, and the obtained analytical sample was appropriately diluted, and the peptide was quantified by LC / MS / MS. At this time, the commercially available whey hydrolyzate was also dissolved in water so as to have a concentration of 10 w / v%, and the same treatment as in the above sample was performed to quantify the peptide. The analysis conditions were in accordance with the conditions shown in Example 1. The results are shown in Table 4 below. The numerical values are expressed as a ratio when the WY concentration in the reaction solution obtained by the enzyme preparation No. 35 (proteolytic enzyme preparation derived from Aspergillus meleus) is set to 100 (displayed value = measured concentration (ppm) / enzyme preparation). WY concentration (ppm) x 100) in the reaction solution obtained by No. 35.

From the results in Table 4, the use of enzyme preparations containing proteolytic enzymes derived from Aspergillus oryzae, Bacillus subtilis or Aspergillus meleus tends to increase the yield of brain function active peptides, especially those derived from Aspergillus meleus. It was found that the use of an enzyme preparation containing the proteolytic enzyme of GTWY and the like increased the yield. It was also found that the whey decomposition product of the present invention has an extremely high concentration of the six peptides of the present invention as compared with the commercially available whey hydrolyzate.

(3) Examination of enzyme reaction conditions The present inventors put protease P "Amano" 3SD into a buffer solution containing 5 to 20 w / v% of various whey proteins at a concentration of 0.05 to 0.2 M and proteolytic enzyme. Add the mixture so that the final concentration of the preparation is 0.1 to 0.5 w / v%, and react the obtained solution at pH 7.0 and 50 ° C. for 1 to 8 hours, and then at 80 ° C. for 30 minutes. It was confirmed that the desired peptide was produced under the condition of inactivating the enzyme (data not shown). It was also confirmed that the reaction proceeded appropriately when the mixture was stirred at 250 rpm (data not shown). Further, the final concentration of the proteolytic enzyme preparation of protease P "Amano" 3SD is 0.2 w / v% in 0.1 M potassium phosphate buffer containing 10 w / v% of whey protein (WPC472 manufactured by Fontera). It was also confirmed that it is preferable to maintain the obtained solution at pH 7.0 and 50 ° C. for 5 to 8 hours and then at 80 ° C. for 30 minutes to inactivate the enzyme (data not shown). ..

Therefore, the enzyme reaction conditions were further investigated for the purpose of further increasing the yield of the specific peptide contained in the whey decomposition product. Specifically, the final concentration of the proteolytic enzyme preparation of protease P "Amano" 3SD is 0.2 w / v in 0.1 M potassium phosphate buffer containing 10 w / v% of whey protein (WPC472 manufactured by Fontera). It was added so as to be%, and the reaction conditions were examined.

The ratio of the peptide concentration of the whey decomposition product obtained by reacting at 50 ° C. for 6 to 8 hours to the WY concentration of 100 when reacting at 50 ° C. for 6 hours (displayed value = measured concentration (displayed value = measured concentration). Table 5 below shows the WY concentration (ppm) × 100) when the reaction was carried out at ppm) / 50 ° C. for 6 hours.

From the results in Table 5, it was found that the peptide concentration tended to increase up to the reaction time of 7 hours, but decreased at 8 hours.

Next, the result when the temperature was changed and maintained after the 6th hour of the reaction was the ratio when the WY concentration when the reaction was carried out at 50 ° C. for 5 hours was 100 (displayed value = measured concentration (ppm)). The WY concentration (ppm) x 100) when reacted at / 50 ° C. for 5 hours is shown in Table 6 below.

From the results in Table 6, it was found that the amount of the obtained peptide was increased by reacting at 50 ° C. for a certain period of time, then reacting at 60 ° C. for a certain period of time, and then inactivating the enzyme at 80 ° C. .. In particular, for WY, it was found that the peptide concentration was increased by reacting at 50 ° C. for 5 hours and then at 60 ° C. for 1 to 3 hours. Furthermore, it was found that when the reaction temperature was raised from 50 ° C. to 60 ° C., the total amount of peptides was maintained even when the total reaction time was 8 hours.

Example 2: Production of peptide degradation product Protease P "Amano" 3SD is added to 0.1 M potassium phosphate buffer (pH 7.0) containing 10 w / v% of whey protein (WPC472 manufactured by Fontera) to complete the proteolytic enzyme preparation. The mixture was added so as to have a concentration of 0.2 w / v%, and the reaction was maintained at 50 ° C. for 5 hours. Then, it was maintained at 80 ° C. for 30 minutes to inactivate the enzyme.

The obtained whey decomposition product-containing reaction solution and the whey decomposition product were freeze-dried at room temperature on the outside of the flask containing the reaction solution using a freeze-dryer FDU-2100 manufactured by EYELA. The peptide concentrations in the powdered whey decomposition product powder are shown in Tables 7 and 8 below.

In this test, 500 g of whey was used as a raw material, and about 2 g of a powdered peptide decomposition product was obtained. It was considered that most of the peptide decomposition products were peptides.

The content ratio of the peptide in the powdered whey decomposition product is considered to be equal to the whey decomposition product-containing aqueous solution (peptide content ratio in the reaction solution) shown in Table 7.

Example 3: Effect of whey decomposition product intake on memory learning function / cognitive function (1) Evaluation method of memory learning function / cognitive function (Y-shaped maze test)
The mouse originally has the property of selecting a route different from the route selected immediately before. Therefore, when the mouse is put into a Y-shaped maze having three arms having the same width, length, etc., it usually enters an arm different from the arm that entered immediately before. The Y-maze test is a test that utilizes the properties of mice to evaluate short-term memory.

In this test, a Y-shaped maze in which three arms with a length of 25 cm, a wall height of 20 cm, and a floor width of 5 cm are connected at an angle of 120 degrees is used in the Y-shaped maze test. Used as a device for.

The order of the moved arms was recorded when the mouse was placed in the tip of one of the arms in the Y-maze and allowed to search freely for 8 minutes. Here, the case where different arms are selected three times in a row and entered is called a voluntary alternation action. The total number of troops entering the arm and the number of voluntary alternation behaviors during the time were counted, and the volatility of volatility of alternation behaviors (%) was calculated using the following formula (1).

自発的交替行動変動率が高いほど、短期記憶が保持されていることを示す。

The higher the volatility of spontaneous alternation behavior, the more short-term memory is retained.

(2) Effect of ingestion of whey decomposition product The whey decomposition product powder obtained in Example 2 was applied to a 6-week-old male CD-1 mouse (obtained from SLC Japan) at a body weight of 0.3 to 3 mg / kg. It was orally administered intragastrically, and 40 minutes later, 0.80 mg / kg body weight of scopolamine hydrochloride (manufactured by SIGMA) was intraperitoneally administered to induce memory impairment. Twenty minutes after intraperitoneal administration of scopolamine, a Y-maze test was performed (Fig. 1). An experiment was conducted with 10 animals in each group, and the average and standard errors were calculated. The result is shown in FIG.

From the results of FIG. 2, no change in the total number of invasions due to whey decomposition product administration was observed, that is, whey decomposition product administration did not affect the amount of activity (Fig. 2B), but whey decomposition product administration was intake-dependent. It was found that the voluntary shift behavior fluctuation rate was increased (Fig. 2A). This means that whey degradation products retain short-term memory.

Example 4: Effect of Ingestion of Synthetic Peptide on Memory Learning Function / Cognitive Function Six synthetic peptides: GTWY, GYGGVSLP, KPTPEGDLEI, WL, WV, in 6-week-old male CD-1 mice (obtained from SLC Japan). WY was orally administered intragastrically at an amount of 0.1 to 3 μmol / kg body weight, and 40 minutes later, 0.80 mg / kg of scopolamine hydrochloride was intraperitoneally administered to induce memory impairment. Twenty minutes after intraperitoneal administration of scopolamine, the Y-maze test described in Example 3 (1) was performed. An experiment was conducted with 10 animals in each group, and the average and standard errors were calculated. The results are shown in FIGS. 3 to 7.

From the results shown in FIGS. 3 to 6, the mice fed with GTWY had a higher rate of voluntary alternation behavior fluctuation than the non-ingestion group in the intake range of 0.1 to 1 μmol / kg body weight, and maintained short-term memory. It turned out that it was done. Similarly, it was found that in GYGGVSLP and KPTPEGDLEI, the voluntary alternation behavior fluctuation rate was higher in the intake range of 0.1 to 1 μmol / kg body weight than in the non-intake group, and short-term memory was retained. In particular, the increase in the volatility of spontaneous alternation behavior by GTWY, GYGGVSLP, and KPTPEGDLEI tended to depend on the peptide intake.

From the results of FIG. 7, in the mice fed with WL, WV, and WY, the voluntary alternation behavior fluctuation rate was dependent on the intake in the intake range of 0.3 to 3 μmol / kg body weight as compared with the non-ingestion group. Was increased, and it was found that short-term memory was retained.

Example 5: Evaluation of Beverages to which Whey Decomposed Products Are Added It is known that acesulfame potassium and sucralose, which are high-intensity sweeteners, have a lower sweetness than sucrose and the like. Therefore, the effect of whey decomposition products on the post-sweetness caused by high-intensity sweeteners was examined.

(1) Preparation of sample beverage Acesulfame potassium (manufactured by MC Food Specialties Inc.) or sucralose (manufactured by Saneigen FFI) was added to ion-exchanged water as a high-sweetness sweetener so as to have the content shown in Table 9. A sample beverage was prepared by adding the whey decomposition product powder obtained in Example 2 to an aqueous solution so as to have the content shown in Table 9.

(2) Sensory evaluation Each sample beverage was subjected to sensory evaluation. Specifically, whether or not the post-sweetness caused by the high-sweetness sweetener can be improved was evaluated according to the following evaluation criteria.
4: The sweetness is extremely refreshing 3: The sweetness is refreshing 2: The sweetness is slightly refreshing 1: The same sweetness as the additive-free sensory evaluation was performed by 6 trained panelists. Each panelist evaluated the score in 0.5 point increments, calculated the average value of the evaluation scores of the 6 panelists, and judged that 2.5 or more was effective.

(3) Evaluation results Table 9 shows the results of sensory evaluation.

From the results in Table 9, in both the acesulfame K aqueous solution and the sucralose aqueous solution, the sweetness trailing effect was most improved when the whey decomposition product concentration was 1 w / v%. However, since this sample beverage does not contain ingredients other than high-sweetness sweeteners and whey decomposition products, bitterness and harshness derived from whey decomposition products can be felt at this concentration, so the overall taste is Sample beverages with concentrations lower than 1 w / v% were superior.

Example 7: Preparation of Whey Decomposed Product with Improved Flavor In this example, improvement of the flavor of the peptide composition by ultrafiltration membrane treatment was examined.

(1) Preparation of sample beverage A sample beverage (sample water 1) in which 1 g of the whey decomposition product powder obtained in Example 2 was dissolved in 500 mL of distilled water and sample water 1 were subjected to an ultrafiltration membrane (fractional molecular weight of 10 kDa, A sample beverage (sample water 2) obtained by filtration treatment with Merck Millipore) was prepared and used for sensory evaluation.

(2) Sensory evaluation Each sample beverage was subjected to sensory evaluation at room temperature. Specifically, five evaluation items: clarity, astringency, bitterness, offensive odor, and whether or not the degree of preference can be improved were evaluated on a five-point scale. Regarding the clarity, the case with the lowest clarity was set to 1, and the case with the highest clarity was set to 5. Regarding the astringency, the case where the astringency was the weakest was set to 1, and the case where the astringency was the strongest was set to 5. Regarding the bitterness, the case where the bitterness was the weakest was set to 1, and the case where the bitterness was the strongest was set to 5. Regarding the offensive odor, the case where the offensive odor was the weakest was set to 1, and the case where the offensive odor was the strongest was set to 5. Regarding the degree of preference, the case with the lowest degree of preference was set to 1, and the case with the highest degree of preference was set to 5.

Sensory evaluation was performed by 4 trained panelists. Each panelist was evaluated with a score in increments of 1, and the average value of the evaluation scores of the four panelists was calculated.

(3) Evaluation results Table 10 shows the results of the sensory evaluation.

From the results in Table 10, it was confirmed that the clarity, astringency, bitterness, offensive odor and palatability of the whey decomposition product solution could be improved by applying the filtration membrane treatment. That is, it was shown that a peptide composition having an improved flavor can be prepared by filtration membrane treatment.

Example 8: Preparation of whey decomposition product-containing beverage with improved storage stability In this example, the storage stability of the whey decomposition product-containing beverage was examined.

(1) Preparation and storage of sample beverage A flavor composition in which the whey decomposition product powder obtained in Example 2 is added to a commercially available sports drink beverage so as to have a concentration of 2 mg / mL, and further has an action of preventing peptide deterioration. As a product, a sample beverage was prepared by adding and mixing various natural product-derived compositions so as to have the contents shown in Table 11. Each of the prepared sample beverages was filled in a 195 mL PET bottle, stored for 1 week, and then used for evaluation. The storage conditions were the temperature and illuminance conditions (preservation condition A and storage condition B) shown in Table 11. The irradiation in the table means that the illuminance during storage was 3000 lux (LX).

(2) Evaluation of Peptide Content The peptide concentration in the sample beverage prepared and stored in (1) above was measured according to the method described in Example 1. Table 12 shows the measurement results for the sample under storage condition A, and Table 13 shows the measurement results for the sample under storage condition B. The peptide concentration is the ratio when the value of sample beverage 1 (without flavor composition added, stored in a dark place) is 100 (displayed value = measured concentration (ppm) / each peptide concentration (ppm) of sample beverage 1). It is shown as × 100).

From the results in Table 12, it was confirmed that the beverages containing the flavor composition (sample beverages 3, 4, 5, 6, 7) suppressed the depletion of the peptide by light. In addition, from the results in Table 13, it was confirmed that the beverages containing the flavor composition (sample beverages 3, 4, 5, 6, 7) suppress the thermal depletion of the peptides. That is, it was shown that a whey decomposition product-containing beverage having improved storage stability can be prepared by adding a flavor composition.

Claims (23)

WY of 0.00001 to 0.2% by mass with respect to total peptide content, WL of 0.000001 to 0.1% by mass with respect to total peptide content, 0.000001 to 0 with respect to total peptide content A peptide comprising one or more components selected from the group consisting of 0.000005 to 0.2% by weight GTWY (SEQ ID NO: 1) with respect to 1% by weight WV and total peptide content. Composition. It further comprises KPTPEGDLEI (SEQ ID NO: 2) and / or GYGGVSLP (SEQ ID NO: 3), and the KPTPEGDLEI content is 0.000001 to 0.05% by weight based on the total peptide content. The peptide composition according to claim 1, wherein the GYGGVSLP content is 0.000005 to 0.2% by mass. The peptide composition according to claim 1 or 2, wherein the total content of WY, WL, WV, GTWY, KPTPEGDLEI and GYGGVSLP is 0.00002 to 0.6% by mass based on the total peptide content. The peptide composition according to any one of claims 1 to 3, which is in the form of a solution. 4. The concentration of WY in the solution is 0.15 ppm or more, the concentration of WL is 0.15 ppm or more, the concentration of WV is 0.1 ppm or more, and the concentration of GTWY is 0.3 ppm or more. The peptide composition according to. The peptide composition according to claim 4 or 5, wherein the GTWY concentration in the solution is 3 ppm or more. The peptide composition according to any one of claims 4 to 6, wherein the KPTPEGDLEI concentration in the solution is 0.1 ppm or more and the GYGGVSLP concentration is 0.1 ppm or more. The peptide composition according to any one of claims 4 to 7, wherein the total of the WY concentration, the WL concentration, the WV concentration, the GTWY concentration, the KPTPEGDLEI concentration and the GYGGVSLP concentration in the solution is 1 ppm or more. The peptide composition according to any one of claims 4 to 8, wherein the total of the WY concentration, the WL concentration, the WV concentration and the GTWY concentration in the solution is 0.5 ppm or more. The peptide composition according to any one of claims 4 to 9, wherein the total of the WY concentration, the WL concentration and the WV concentration in the solution is 0.5 ppm or more. Peptide composition in ingestion form comprising one or more components selected from the group consisting of 160-70000 μg WY, 100-6000 μg WV, 100-6000 μg WL and 50-40,000 μg GTWY. Peptide composition consisting of. The peptide composition according to claim 11, wherein the content of GTWY is 100 to 30,000 μg. The peptide composition according to claim 11 or 12, further comprising GYGGVSLP and / or KPTPEGDLEI, wherein the content of GYGGVSLP is 50,000 to 50,000 μg and the content of KPTPEGDLEI is 160 to 70,000 μg. The peptide composition according to any one of claims 1 to 13, which is in the form of a food. The composition according to claim 14, wherein the food is a beverage. A food product comprising the peptide composition according to any one of claims 1 to 13. The food product according to claim 16, which is a beverage. The peptide composition according to any one of claims 1 to 15 or the food according to claim 16 or 17, for maintaining, improving and / or improving brain function. An effective amount of the peptide composition according to any one of claims 1 to 15 or the food according to claim 16 or 17 is ingested or administered to a human or non-human animal. How to maintain brain function, how to improve brain function and how to improve brain function. Use of the peptide composition according to any one of claims 1 to 15 for the production of compositions and foods for maintaining, improving and / or improving brain function. The peptide composition according to any one of claims 1 to 15, comprising a step of reacting a whey protein with an enzyme preparation containing one or more kinds of proteolytic enzymes derived from Aspergillus meleus to obtain an enzymatically decomposed product. How to make things. The production method according to claim 21, wherein the step of allowing the enzyme to act on the whey protein is carried out by holding the whey protein at an action temperature of 45 to 55 ° C. for a certain period of time and then holding the whey protein at an action temperature of 60 to 75 ° C. for a certain period of time. The production method according to claim 21 or 22, further comprising a step of diluting an enzymatic degradation product of whey protein or a step of concentrating an enzymatic degradation product of whey protein.

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