JP2008056645A - Anti-oxidant peptide obtained by reaction of protein in enzymatically treated royal jelly with polypeptide and method for producing the same - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an anti-oxidant peptide obtained by reacting a polypeptide with a protein in a novel royal jelly which can impart to cells anti-oxidant properties and anti-aging properties as well as effectiveness inherent in a water-solubilized royal jelly (having a lowered molecular weight and suspended but apparently dissolved in water) and to provide a method for producing the same. <P>SOLUTION: What are provided are an anti-oxidant peptide obtained by reacting a polypeptide with a protein in an enzymatically treated royal jelly and having a molecular weight falling within the range of 100 to 1,000 and a method for producing the same. <P>COPYRIGHT: (C)2008,JPO&INPIT

Description

  The present invention relates to an antioxidant peptide obtained by a reaction between a protein and a polypeptide in an enzyme-treated royal jelly and a method for producing the same.

  Royal jelly is a milky white, viscous and sour substance secreted from the pharyngeal glands of worker bees, and is well known to be given to all bee larvae for the first few days, but only to larvae that become queen bees thereafter. Among these, nutrients such as vitamins, minerals, amino acids, acetylcholine, 10-hydroxydecenoic acid, sterols, and hormones are included in a well-balanced manner, and are used in fields such as health foods, pharmaceuticals, and cosmetics. In Patent Document 1, this royal jelly is made water-soluble, thereby succeeding in expanding its utility. However, this is not satisfactory in terms of antioxidant properties and anti-aging properties.

Japanese Patent No. 2623044

The first object of the present invention is to impart antioxidant and anti-aging properties to cells in addition to the effects of water-solubilized royal jelly (which appears to be dissolved in a state of low molecular weight floating in water). The object is to provide an antioxidant peptide obtained by reacting a protein in a novel royal jelly with a polypeptide.
The second object of the present invention is to provide a method for producing an antioxidant peptide obtained by reacting a protein in a novel royal jelly with a polypeptide.

The first of the present invention relates to an antioxidant peptide obtained by reacting a protein in a royal jelly treated with an enzyme and a polypeptide and having a molecular weight distribution in the range of 100 to 1,000.
The second of the present invention is an antioxidant peptide obtained by reacting a protein in a royal jelly treated with an enzyme and a polypeptide, the molecular weight distribution of which is from 100 to less than 500-70% to 90%
30 to 10% of those less than 500 to 1000 (where% is a percentage of peak area)
It is related with the antioxidant peptide of Claim 1.
3rd of this invention is related with the antioxidant peptide of Claim 1 or 2 obtained by making 0.7 weight part or more of polypeptides react with 100 weight part of royal jelly processed with an enzyme.
The fourth aspect of the present invention is that the enzyme-treated protein in the royal jelly is reacted with the polypeptide in a buffer solution of pH 4-6 at a temperature of 60 ° C. or lower. The present invention relates to a method for producing an antioxidant peptide.

The enzyme-treated royal jelly in the present invention can be obtained by the method described in Patent Document 1. That is, the enzyme-treated royal jelly is “preparing an aqueous suspension of royal jelly, and simultaneously adding two or more proteases having different action sites to the substrate to this, and maintaining the temperature at room temperature or higher, After solubilizing insoluble substances by reaction, the enzyme reaction is stopped by heating to 60 ° C. or higher at pH 5.0 to 6.0, and then the precipitate is removed to produce in the form of a clear royal jelly solution. be able to.
In order to adjust the pH in this way, an alkali such as NaOH or KOH, or an acid such as citric acid, gluconic acid or malic acid or an organic acid such as a dibasic amino acid (arginine, lysine, etc.) is used. Since the aqueous suspension of royal jelly is often very acidic, alkali is often used for pH adjustment. When an alkali metal hydroxide such as NaOH or KOH is used, alkali metal always remains, so use of KOH is preferable in consideration of a sodium restrictor such as kidney disease.
Without such enzyme-treated royal jelly, no reaction occurs with the polypeptide. In addition, what is substantially reacted with the polypeptide is a protein in the royal jelly treated with an enzyme, and other components are not reacted and are present in the reaction product as they are.

  The polypeptide in the present invention is preferably a biological enzyme polypeptide. The polypeptide is preferably one having 10 to 100 amino acids in the molecular structure and having a molecular weight of 500 to 1500. Biological enzymes include metabolic enzymes and digestive enzymes, but biological enzyme polypeptides belong to metabolic enzymes. Specific examples thereof include SOD (Super Oxide Dismutase Enzyme), DNA bricosylase, AP endonuclease, DNA helicase, DNA polymerase, DNA ligase, E2-type protein, P53 protein, P21 protein, telomerase, glycerologheptid α D-glycoheptite), D-glycero-D-idheptit (β-D-glycoheptit), L-glycero-L-idheptit (β-L-glycoheptit), D-glycero-D-galaheptit (α -D-Mannoheptit), L-glycero-L-galaheptit (α-L-Mannoheptit), D-glycero-D-Mannoheptit (β-D-Mannoheptit), D-Glycero-D- Glucoheptite (β-D-Altolohe Chit), L-glycero-L-glucoheptite (β-L-altoheptit), D-glycero-L-glucoheptit (β-D-galaheptit), L-glycero-D-glucoheptit (β -L-galaheptit), glyceroid heptit (β-idheptit), glyceroalloheptit (β-alloheptit), D-glycero-D-altoheptit (α-D-alloheptit) and the like.

  The reaction between the protein and the polypeptide in the enzyme-treated royal jelly is usually a first step of mixing the pH 5.0 to 6.0 while adjusting the pH to 5.0 to 6.0 and a temperature of 60 ° C. or lower. The second step of proceeding the reaction while adjusting to 0 and a temperature of 60 ° C. or lower, heating to 60 to 80 ° C. while adjusting to pH 5.0 to 6.0, and then cooling to 20 ° C. or lower, insoluble matter It consists of the 3rd process to remove. These steps are usually completed within 1 hour. The pH is adjusted with a phosphate buffer solution having a pH of 4 to 6, for example.

  Since the reaction product obtained by the reaction step is in the form of an aqueous solution, it may be used as it is for the intended purpose. However, the reaction product may be dried as it is or the antioxidant peptide in the reaction product. Only can be decomposed and dried to form a solid. As a drying means, it can carry out using carriers, such as powder and sugar, or freeze drying, spray drying, etc.

  The quantitative relationship between the protein and the polypeptide in the enzyme-treated royal jelly is 0.7 parts by weight or more, preferably 0.7 to 2.0 parts by weight of the polypeptide with respect to 100 parts by weight of the enzyme-treated royal jelly. Particularly preferred is 1.2 to 1.4 parts by weight. When the amount is less than 0.7 parts by weight, the antioxidant effect is insufficient. Even if it is 2.0 parts by weight or more, there is no change in the active efficacy.

  In the reaction product of the present invention or the production method of the present invention, a third component such as a flavonoid or a coenzyme can be present if necessary. Flavonoids include anthocyanins, flavans, and isoflavonoids. Common names include polyphenol, catechin, astaxanthin and the like. Examples of coenzymes include vitamin coenzymes and quinone coenzymes. Specific examples include pantothenic acid, vitamin B12, and coenzyme Q.

The royal jelly used as a raw material for producing the enzyme-treated royal jelly is not limited to a raw one, and any one such as a frozen one or a freeze-dried one can be used as necessary.
First, a royal jelly suspension is prepared by adding water or warm water to the royal jelly. In this case, pretreatment with alcohol (eg, ethanol) etc. (filtering and purification) or preparing a suspension using an alcohol solution will reduce the recovery rate of useful components in royal jelly, especially proteins insoluble in alcohol. Therefore, it is not preferable to perform such pretreatment or solubilization treatment with an alcohol solution.

  The royal jelly suspension with adjusted pH is subjected to protein degradation by adding protease (proteolytic enzyme). There are no particular restrictions on the protease used, such as digestive enzymes derived from mammals such as acidic proteases, neutral proteases, alkaline proteases from microorganisms and plants, pepsin, pancreatin, etc., which are usually used in food processing Can be widely used, and two or more types having different action sites for the substrate can be selected and used. Specific examples of the protease (proteolytic enzyme) include carboxypeptidase, aminopeptidase, dipeptidase, pepsin, trypsin, chymotrypsin, rennin, pancreatin, papain, and chymopapain.

  The enzyme reaction (enzyme treatment) can be arbitrarily set by changing the kind and amount of enzyme, reaction temperature and reaction time, but the usual treatment is 20 to 500 units / g of enzyme per 1 kg of royal jelly. 0.5 to 2 g is added and treated for 2 to 24 hours. Although reaction temperature can be performed at 20-60 degreeC, it is desirable to set it as 40-50 degreeC from the point of browning prevention or decomposition efficiency.

When enzyme treatment is performed with two or more types of proteases, if the preferred pH conditions and temperature conditions of the respective enzymes are relatively close to each other, a plurality of types of enzymes can be added and treated at the same time. It is convenient because the operation is simple and a relatively high decomposition rate can be obtained. The degradation rate here refers to the rate at which proteins are degraded and the molecular weight is reduced. On the other hand, when the enzyme treatment is performed by combining enzymes having different preferable pH conditions and temperature conditions for each enzyme, sequential reactions are performed. In this case, the treatment conditions can be set to preferable conditions for each enzyme, and an enzyme-treated royal jelly solution in which various kinds of proteases are combined can be obtained. Enzyme treatment measures the degradation rate of protein over time, and is terminated when the degradation rate reaches the maximum. Although the final decomposition rate varies depending on the enzyme used, the reaction is terminated when the decomposition rate is 75% or more, preferably 85% or more, and the enzyme is deactivated by heating at a temperature of 60 to 100 ° C. for 5 to 30 minutes. Let After stopping the enzyme reaction, impurities or insoluble components are removed by centrifugation or membrane filtration to obtain a transparent royal jelly solution.
By combining two or more kinds of enzymes in this way, the rate of protein degradation in royal jelly is dramatically increased compared to single enzyme treatment, and the stability when this royal jelly is added to an acidic beverage is increased. The property is extremely excellent. When royal jelly is treated with protease, a generally preferred flavor and umami are also added. It is important to choose a good enzyme combination in terms of taste.
By the way, in Example 1, royal jelly was followed by pepsin treatment followed by pancreatin treatment. In Example 2, pepsin treatment was followed by neutral protease treatment. In Example 3, pepsin treatment was followed. Although processing is performed, this order is not particularly limited. By these enzyme treatments, the molecular weight of the protein is subdivided and peptide formation is promoted.

The enzyme inactivation treatment by heating the royal jelly suspension after the enzyme treatment is carried out at 60 ° C. or higher at pH 5.0 to 6.0, and the enzyme reaction is stopped, followed by precipitation by membrane filtration or centrifugation. Remove the material to make a clear royal jelly solution. This royal jelly is an enzyme-treated royal jelly used in the present invention.
By stopping the enzymatic reaction within this range, the recovery rate of crude protein and total lipid can be increased, and in particular, the recovery rate of 10-hydroxydecenoic acid, which is an active ingredient in royal jelly, can be significantly increased. The dissolution stability of useful components can be increased.

An antioxidant peptide having a molecular weight distribution in the range of 100 to 1000 obtained by the reaction of the protein and polypeptide in the enzyme-treated royal jelly of the present invention,
70-90% having a molecular weight of less than 100-500, preferably 75-90%,
30 to 10% having a molecular weight of less than 500 to 1000, preferably 25 to 10%,
(However,% is the percentage of peak area)
Those having a molecular weight of 1000 or more are hardly contained.
On the other hand, the protein in the enzyme-treated royal jelly (in Comparative Example 1 described later)
24% * with a molecular weight of 10,000 or more
20% with a molecular weight of 3000 to less than 10,000 *
Molecular weight 1000 to less than 3000 Trace amount Molecular weight 500 to less than 1000 Trace amount Molecular weight Less than 100 to 500 56% *
It is. * Indicates that these contents vary somewhat depending on the amount of trace components having a molecular weight of 1000 to less than 3000.
The molecular weight distribution was determined by size exclusion chromatography using a TSKgel G2600PWxl column.

前記表１から明らかなように、本発明方法により得られたローヤルゼリー／ポリペプチド反応生成物溶液は、特許文献１の製造方法により得られたローヤルゼリー溶液と比較して、抗酸化ペプチドの量が２〜３倍量であり、デセン酸は４〜８倍量であるのに対して、ナトリウム量が１／１０程度と少ない。 The contents of decenoic acid, antioxidant peptide, and sodium in the royal jelly / polypeptide reaction product solution obtained by the method of the present invention and the royal jelly solution obtained by the method of Patent Document 1 are generally as shown in the following table. .

As is apparent from Table 1, the amount of antioxidant peptide in the royal jelly / polypeptide reaction product solution obtained by the method of the present invention is 2 in comparison with the royal jelly solution obtained by the production method of Patent Document 1. The amount of sodium is about 1/10, whereas the amount of decenoic acid is 4 to 8 times.

The present invention is not a simple mixture of an enzyme-treated royal jelly and a polypeptide, but a reaction product of both, and is an anti-oxidation peptide mainly having a tetramer (tetramer) structure, and thus has not reacted. Peptide activity (enhances absorption in the small intestine) and enzyme activity (enhances degradation and metabolic capacity) are improved 4 to 10 times compared to a simple mixture, an enzyme-treated royal jelly alone, or a polypeptide alone (for example, FIG. 1).
The antioxidant peptide obtained by the reaction of the protein and the polypeptide in the royal jelly treated with the enzyme of the present invention exhibits an anti-aging action and an antioxidant action, promotes the proliferation of human blood cell monocytes, It has cholesterol-reducing action, neutral fat-reducing action, and visceral fat mass-reducing action.
There are two types of cell death, one of which is necrosis (necrosis) and the other is apoptosis. The former is death with fever and inflammation because the cells swell and collapse, while the latter is death without fever and inflammation while the cells shrink and are enveloped in the cell membrane. The reaction product of the enzyme-treated royal jelly and the polypeptide of the present invention causes apoptosis of cancer cells, but has no effect on normal cells. Rather, it promotes proliferation and growth for normal cells.
The antioxidant peptide obtained by the reaction of the protein and polypeptide in the enzyme-treated royal jelly of the present invention enhances immunity.
The antioxidant peptide obtained by the reaction between the protein and the polypeptide in the enzyme-treated royal jelly of the present invention is useful as a food additive intended for anti-aging or anti-cancer action, skin care cosmetics and the like.

  Hereinafter, the present invention will be described with reference to examples, but the present invention is not limited thereto.

Example 1
(1) Production of Royal Jelly Treated with Enzyme The pH of the 10 wt% royal jelly aqueous suspension was adjusted to 4 using a 20 wt% potassium hydroxide solution. 1 g of pepsin was added to this solution, and enzyme treatment was performed at 45 ° C. for 6 hours (pepsin treatment solution). Next, the pH of this treatment solution was adjusted to 8 using a 20 wt% potassium hydroxide solution, 1 g of pancreatin was added, and enzyme treatment was performed at 45 ° C. for 6 hours. The enzyme-treated solution is adjusted to pH 5.5 with a 6 wt% or more decenoic acid solution, heated at 80 ° C. for 10 minutes to deactivate the enzyme, and further filtered to remove foreign matters and insoluble residues. A clear royal jelly solution was obtained. This royal jelly solution was concentrated under reduced pressure to the weight of raw royal jelly.
(2) Reaction of protein and polypeptide in enzyme-treated royal jelly A raw material obtained by mixing 9 parts by weight of royal jelly concentrate containing 6 wt% decenoic acid obtained in the step (1) and 1 part by weight of SOD polypeptide The pH was adjusted to 4 with 1 kg of 10 wt% aqueous suspension using 20 wt% potassium hydroxide solution. 1 g of pepsin was added to this solution, and enzyme treatment was performed at 45 ° C. for 6 hours (pepsin treatment solution). Next, the pH of this treatment solution was adjusted to 8 using a 20 wt% potassium hydroxide solution, 1 g of pancreatin was added, and enzyme treatment was performed at 45 ° C. for 6 hours. The solution after completion of the enzyme treatment was adjusted to pH 5.5 using a 20 wt% potassium hydroxide solution or a 10 wt% citric acid solution and heated at 80 ° C. for 10 minutes to deactivate the enzyme (the purpose of the enzyme is terminated) Then, it is important to inactivate), and further filtration was performed to remove foreign substances and insoluble residues to obtain a solution of a transparent royal jelly polypeptide reaction product (including antioxidant peptide). This royal jelly polypeptide reaction product (including antioxidant peptide) solution was concentrated under reduced pressure to the weight of the raw royal jelly used to obtain a product.

Example 2
The pH of the pepsin treatment solution 1 kg was adjusted to 7 using a 20 wt% potassium hydroxide solution. 1 g of neutral protease was added to this solution, and enzyme treatment was performed at 45 ° C. for 6 hours. The solution after completion of the enzyme treatment is adjusted to pH 5.5 using 20 wt% potassium hydroxide solution or 10 wt% citric acid solution, heated at 80 ° C. for 10 minutes to deactivate the enzyme, and further filtered to remove foreign matter. In addition, a transparent royal jelly polypeptide reaction product (containing antioxidant peptide) solution was obtained. This royal jelly polypeptide reaction product (including antioxidant peptide) solution was concentrated under reduced pressure to the weight of the raw royal jelly used to obtain a product.

Example 3
20 wt% water of 1 kg of a 10 wt% royal jelly water suspension of a raw material obtained by mixing 9 parts by weight of royal jelly containing 9 wt% of decenoic acid obtained by the method of Example 1 (1) and 1 part by weight of SOD polypeptide The pH was adjusted to 4 using a potassium oxide solution. 1 g of acidic protease was added to this solution, and enzyme treatment was performed at 45 ° C. for 6 hours (acid protease treatment solution). 1 g of pepsin was added to this treatment solution, and enzyme treatment was performed at 45 ° C. for 6 hours. The solution after completion of the enzyme treatment is adjusted to pH 5.5 using 20 wt% potassium hydroxide solution or 10 wt% citric acid solution, heated at 80 ° C. for 10 minutes to deactivate the enzyme, and further filtered to remove foreign matter. In addition, a transparent royal jelly polypeptide reaction product (containing antioxidant peptide) solution was obtained. This royal jelly polypeptide reaction product (including antioxidant peptide) solution was concentrated under reduced pressure to the weight of the raw royal jelly used to obtain a product.

Comparative Example 1
The pH was adjusted to 4 using 20 wt% potassium hydroxide solution of 1 kg of 10 wt% royal jelly water suspension. 1 g of pepsin was added to this solution, and enzyme treatment was performed at 45 ° C. for 6 hours (pepsin treatment solution). Next, the pH of this treatment solution was adjusted to 8 using a 20 wt% potassium hydroxide solution, 1 g of pancreatin was added, and enzyme treatment was performed at 45 ° C. for 6 hours. The solution after completion of the enzyme treatment is adjusted to pH 5.5 using 20 wt% potassium hydroxide solution or 10 wt% citric acid solution, heated at 80 ° C. for 10 minutes to deactivate the enzyme, and further filtered. Foreign substances and insoluble residues were removed to obtain a transparent royal jelly solution.

２）試験溶液の調製
それぞれ検体約０．０１ｇを採取し、水、アセトニトリル及びトリフルオロ酢酸の混液（５４．９：４５：０．１）１０ｍｌを加えた。室温で一晩放置し充分に溶解させた後、孔径０．４５μｍのメンブランフィルターでろ過し、得られた液を試験溶液とした。
３）分子量分布の測定
標準溶液及び試験溶液について、サイズ排除カラム（ＴＳＫｇｅｌ Ｇ２６００ＰＷｘｌカラム）を用いる高速液体クロマトグラフ装置に注入した。得られた結果を４８０IIデータステーションＧＰＣプログラム（システムインスツルメンツ株式会社）を用いて解析した。なお、各ピークの分子量の推定を分子量標準品の溶出時間及び分子量をもとに作成した検量線を用いて行った。
（３）試験結果
検体の分子量分布測定結果を表２に示した。

(1) The molecular weight distribution of the antioxidant peptide of Example 1 (Sample 1) and the protein in the enzyme-treated royal jelly (Sample 2) of Comparative Example 1 were measured.
(2) Test method 1) Preparation of standard solution The molecular weight standard products shown in Table 1 below were dissolved in a mixed solution of water, acetonitrile and trifluoroacetic acid (54.9: 45: 0.1) to 0.05 to 0. A 1 W / V% solution was prepared.

2) Preparation of test solution About 0.01 g of each specimen was collected, and 10 ml of a mixed solution of water, acetonitrile and trifluoroacetic acid (54.9: 45: 0.1) was added. The mixture was allowed to stand at room temperature overnight for sufficient dissolution, and then filtered through a membrane filter having a pore size of 0.45 μm, and the resulting solution was used as a test solution.
3) Measurement of molecular weight distribution The standard solution and the test solution were injected into a high-performance liquid chromatograph using a size exclusion column (TSKgel G2600PWxl column). The obtained results were analyzed using a 480II data station GPC program (System Instruments Inc.). The molecular weight of each peak was estimated using a calibration curve prepared based on the elution time and molecular weight of a molecular weight standard product.
(3) Test results Table 2 shows the molecular weight distribution measurement results of the specimens.

Test example 1
Antioxidant SOD activity was measured by the cytochrome C method for comparison between the royal jelly polypeptide reaction product solution shown in Example 1 and the royal jelly solution obtained by the production method of Comparative Example 1. In this measurement method, O ⁻² (active oxygen) generated in the xanthine oxidase system is detected by cytochrome C reduction (absorption increase at 550 nm), and O ⁻² (active oxygen) is eliminated by the added SOD sample. The amount of SOD in the sample is measured from the inhibition rate of cytochrome C reduction. According to the known method of McCord and Fridovich (November 6, 1989, published by Nippon Medical Center Co., Ltd., Yoshihiko Oyanagi “SOD and “Refer to“ Method for Measuring Active Oxygen ”in“ Active Oxygen Regulator—Pharmacological Action and Clinical Application ”).

Test method Sample A: Royal jelly polypeptide reaction product solution sample B shown in Example 1 Royal jelly solution sample C obtained by the production method described in Example 1 (1) C: Raw royal jelly (untreated Royal jelly)
In each of the three containers, 0.1 ml of 0.3 mM cytochrome C solution and 0.2 ml of 1.5 mM xanthine solution were added to 2.2 ml of 50 mM phosphate buffer (pH 7.8) containing 0.1 mM ethylenediaminetetraacetic acid (EDTA). In addition, sample A is added to one of the containers, sample B is added to the other container, and 0.1 ml of sample C is added to the other container to start the reaction. Incubate at 25 ° C for 5 minutes. Add 0.1 ml of xanthine oxidase solution to each container and carry out the reaction. The change in absorbance at 550 nm at 25 ° C. is measured with a spectrometer for about 2 minutes.
The amount of SOD that inhibits the reduction rate of cytochrome C by the above enzyme reaction by 50% was defined as 1 unit.
The antioxidative SOD activities of the obtained samples A, B, and C are as shown in FIG. 1. The antioxidative SOD activity of the present invention is 4 to 10 times that of raw royal jelly and enzyme-treated royal jelly. Is shown.

Test example 2
Table 4 shows a component comparison of the royal jelly polypeptide reaction product solution A shown in Example 1 and the royal jelly solution B obtained by the production method of Example 1 of ordinary Patent Document 1.

Test example 3
Using the royal jelly polypeptide reaction product solution shown in Example 1, it was confirmed that it had an apoptotic action as shown in FIG. There are two types of cell death, one of which is necrosis (necrosis) and the other is apoptosis. The former is death with fever and inflammation because the cells swell and collapse, while the latter is death without fever and inflammation while the cells shrink and are enveloped in the cell membrane. The reaction product of the enzyme-treated royal jelly and the polypeptide of the present invention causes apoptosis of cancer cells, but has no effect on normal cells. Rather, it promotes proliferation and growth for normal cells. When human blood cell monocytes are cultured in a petri dish and the royal jelly polypeptide reaction product solution shown in Example 1 is added, the royal jelly solution obtained in Example 1 of Patent Document 1 is added and Compared. As shown in FIG. 2, when the number of cells was examined, the royal jelly polypeptide reaction product solution shown in Example 1 was increased about 7 times 5 days after the addition, and the royal jelly solution obtained in Example 1 of Patent Document 1 was increased. Is added to only about 3.5 even after 4 days. From this result, it can be seen that the royal jelly polypeptide reaction product solution shown in Example 1 has an action of promoting cell proliferation.
Next, the case of FIG. 3 in which murine liver cells are cultured in the same manner will be described. Those who added the royal jelly polypeptide reaction product solution shown in Example 1 had doubled the number of cells after one day, and about 90% of the cells remained alive even after 20 days. On the other hand, the person who added the royal jelly solution obtained in Example 1 of Patent Document 1 increased 1.8 times after 1 day, but after that, the cell suddenly died and after 20 days it decreased to the same level as before the start of culture. I have. From this result, it was found that the royal jelly polypeptide reaction product solution shown in Example 1 did not have a strong growth promoting effect on liver cells, but had the effect of allowing the increased number of cells to live longer. This is thought to prevent the liver cells from dying by the mechanism of apoptosis.

Test example 4
As shown in FIG. 4 (A), the royal jelly polypeptide reaction product solution shown in Example 1 was tested in the same manner as in Test Example 3, and the rat bad cholesterol (LDL: low density) that ate a high fat diet. Although the polyprotein was decreased, as shown in FIG. 4B, it was found that it had no effect on good cholesterol (HDL: high density polyprotein). In the same experimental system as FIG. 4, it was found that the neutral fat of the mouse is lowered (FIG. 5), and further, there is an effect of reducing the fat mass around the internal organs (FIG. 6). Moreover, as shown in FIG. 1 by the same experimental system as FIGS.

(A) When the royal jelly polypeptide reaction product of Example 1 is added, (B) When the enzyme-treated royal jelly solution obtained by the method of (1) of Example 1 is added, and (C) Raw royal jelly The duration of each antioxidant effect in the case where is added is shown. The cell proliferation effect when human blood cell single cells are cultured in a petri dish and the royal jelly polypeptide reaction product of Example 1 is added thereto is shown. The cell growth effect is shown when murine liver cells are cultured in a petri dish and the royal jelly polypeptide reaction product of Example 1 is added thereto. (A) The result of having investigated the density | concentration of bad cholesterol (LDL) in the case where the royal jelly polypeptide reaction product of Example 1 is given to the rat eating a high fat diet and in the case where casein is given. (B) It is the result of having investigated the density | concentration of good cholesterol (HDL) similarly. The royal jelly polypeptide reaction product of Example 1 shows that it has a neutral fat lowering ability as compared with casein. The royal jelly polypeptide reaction product of Example 1 shows that it has the ability to reduce visceral fat mass compared to casein.

Claims (4)

  An antioxidant peptide obtained by a reaction between a protein and a polypeptide in royal jelly treated with an enzyme and having a molecular weight distribution in the range of 100 to 1,000. 70-90% of an antioxidant peptide obtained by reacting a protein in a royal jelly treated with an enzyme and a polypeptide, the molecular weight distribution of which is less than 100-500
30 to 10% of those less than 500 to 1000 (where% is a percentage of peak area)
The antioxidant peptide according to claim 1.   The antioxidant peptide according to claim 1 or 2, obtained by reacting 0.7 parts by weight or more of a polypeptide with 100 parts by weight of royal jelly treated with an enzyme. 4. The method for producing an antioxidant peptide according to claim 1, wherein the enzyme and the protein in the royal jelly are reacted in a buffer aqueous solution having a pH of 4 to 6 under a temperature condition of 60 ° C. or lower. .

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