JP2006061038A - Enzyme-treated royal jelly-containing health drink - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an enzyme-treated royal jelly(RJ)-containing health drink having high quality and easily controlling the generation of insoluble substances. <P>SOLUTION: This enzyme-treated royal jelly-containing health drink is obtained by making water contain RJ (enzyme-treated RJ) subjected to protein decomposition enzyme treatment using an endo-type neutral protease derived from Bacillus subtilis, and gellan gum as a polysaccharide thickener. It is preferable that the content of the enzyme-treated RJ in the health drink is 0.6-1.2 mass%, and the content of the gellan gum in the health drink is 0.01-0.09 mass%, more preferably 0.02-0.05 mass%. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

  The present invention relates to an enzyme-treated royal jelly-containing drink useful as health drinks, pharmaceuticals, quasi drugs, and the like for the purpose of prevention and treatment of hypertension.

Royal jelly is a natural product that has been conventionally used as a healthy food and drink or herbal medicine with high nutritional value. Royal jelly has blood pressure lowering action, antitumor action, wound healing promotion, serum cholesterol lowering action, blood flow increasing action, sexual central aging prevention action, antibacterial action, growth promoting action, side effect reducing action of radiation and chemotherapeutic agents, chronic diseases It has various physiological actions such as an action on urine and has been reported in many literatures. For example, Non-Patent Document 1 discloses an enzyme-treated royal jelly obtained by treating a protein contained in royal jelly with a proteolytic enzyme.
Hiroe Maruyama, 6 others, “Isolation and identification of angiotensin I-converting enzyme inhibitory peptide contained in protease-treated royal jelly”, Shokuhin, 2003, 50, 310-315.

  However, when royal jelly is used in various beverages such as health drinks, pharmaceuticals, and quasi-drugs, proteins and peptides contained in the royal jelly are aggregated or precipitated by heat sterilization during the production of the beverage. There is a possibility of doing. The above-mentioned conventional royal jelly generally has relatively good solubility in water because the protein is decomposed. However, aggregation and precipitation of proteins and peptides can be avoided as the amount of added royal jelly increases. Absent. In particular, proteins and peptides containing a high amount of hydrophobic amino acids are prone to aggregation and precipitation, and there is a problem that the quality of beverages is likely to be reduced.

  Therefore, as a result of intensive studies, the present inventors have developed a high-quality drink agent in which aggregation and precipitation of proteins, peptides and the like are suppressed, and have completed the present invention. An object of the present invention is to provide a high-quality enzyme-treated royal jelly-containing drink that can easily suppress the formation of insoluble matter.

  In order to achieve the above object, the enzyme-treated royal jelly-containing drink according to claim 1 is an enzyme-treated royal jelly that has been subjected to proteolytic enzyme treatment using an endo-type neutral protease derived from Bacillus subtilis. And containing gellan gum.

  The enzyme-treated royal jelly-containing drink having the above structure contains gellan gum that is particularly excellent in the thickening effect. For this reason, this drink is given suitable viscosity, and as a result, precipitation and aggregation of proteins and peptides contained in the enzyme-treated royal jelly are effectively suppressed. In particular, gellan gum is very excellent in the effect of suppressing precipitation and aggregation of proteins and peptides in enzyme-treated royal jelly. That is, according to this structure, the high quality drink agent by which precipitation and aggregation of protein, a peptide, etc. are suppressed and the production | generation of the insoluble matter resulting from them is suppressed is provided.

  The enzyme-treated royal jelly-containing drink according to claim 2 is an enzyme-treated royal jelly, gellan gum, sugar solution, acidulant, and vitamin C other than vitamin C, which has been subjected to an endo-type neutral protease treatment derived from Bacillus subtilis. It consists of a fragrance, a pigment and water.

  The enzyme-treated royal jelly-containing drink having the above-described configuration does not contain vitamin C (ascorbic acid) that induces browning. Vitamin C is easily oxidized and changes to a substance that easily reacts with amino groups of proteins and peptides to turn brown. Therefore, according to this structure, the production | generation of the insoluble matter resulting from precipitation and aggregation of protein, a peptide, etc. is suppressed, and the high quality drink agent by which the brown change was suppressed is provided.

  The enzyme-treated royal jelly-containing drink of the invention described in claim 3 is the invention according to claim 1 or claim 2, wherein the enzyme-treated royal jelly contains 0.6 to 1.2% by mass, and the gellan gum is 0%. The content is 0.01 to 0.09% by mass.

  The enzyme-treated royal jelly-containing drink having the above structure contains gellan gum in an amount that effectively suppresses precipitation and aggregation of proteins and peptides contained in the enzyme-treated royal jelly. For this reason, the high quality drink agent with which the production | generation of the insoluble matter resulting from precipitation and aggregation was suppressed is provided.

The enzyme-treated royal jelly-containing drink of the invention described in claim 4 is characterized in that it has a blood pressure lowering action in the invention described in any one of claims 1 to 3.
According to the enzyme-treated royal jelly-containing drink having the above-described configuration, a good blood pressure lowering effect is exhibited through the enzyme-treated royal jelly having ACE inhibitory activity.

  ADVANTAGE OF THE INVENTION According to this invention, the high quality enzyme-treated royal jelly containing drink agent which can suppress the production | generation of an insoluble matter easily can be provided.

  Hereinafter, an embodiment in which the enzyme-treated royal jelly-containing drink (hereinafter referred to as drink) of the present invention is embodied will be described in detail. In the present embodiment, “royal jelly” is abbreviated as “RJ”.

  The drink of this embodiment contains an enzyme-treated RJ that has been subjected to a proteolytic enzyme treatment using an endo-type neutral protease derived from Bacillus subtilis, and gellan gum as a thickening polysaccharide. This drink is configured so that the poorly water-soluble component in the enzyme-treated RJ exhibits high dispersion stability with respect to water by the emulsifying action of gellan gum, effectively suppressing the generation of insoluble matter. It is supposed to be. Examples of usage forms of the drink of this embodiment include pharmaceuticals, quasi drugs, health drinks, and the like.

  Furthermore, this drink exhibits a high blood pressure lowering action by inhibiting information transmission related to the blood pressure increase of the renin-angiotensin-aldosterone system by the action of the active ingredient contained in the enzyme-treated RJ. The renin-angiotensin system (RAS), together with catecholamine and vasopressin, plays an important role in regulating blood pressure and maintaining body electrolytes in vivo. That is, angiotensin I produced from angiotensinogen becomes angiotensin II by angiotensin converting enzyme (ACE). Angiotensin II has a strong pressor activity and promotes the secretion of aldosterone from the adrenal gland layer. This angiotensin II exerts pressor activity mainly through a vasoconstrictive action that raises blood pressure by constricting arteriole smooth muscle and an action that acts on the adrenal globular layer to promote synthesis and release of aldosterone. . The aldosterone has an action of increasing sodium reabsorption from the distal renal tubule. On the other hand, the enzyme-treated RJ as an active ingredient contained in the drink has an ACE inhibitory activity and also has an action of inhibiting the secretion of the aldosterone, and suppresses the reabsorption of sodium to lower the circulating plasma volume. As a result, the blood pressure lowering effect is exhibited.

  The enzyme-treated RJ contains at least active ingredients composed of various peptides such as Ile-Tyr (IY), Val-Tyr (VY), and Ile-Val-Tyr (IVY). In addition, the enzyme-treated RJ includes peptides having the amino acid sequences represented by SEQ ID NOs: 1 to 3, respectively. All of the peptides consisting of the amino acid sequences represented by SEQ ID NOs: 1 to 3 function as an active ingredient of a drink, and are excellent in blood pressure lowering action through inhibition of ACE activity, for example. The drink contains the RJ-derived peptide and other unidentified components (mainly peptides) as active ingredients.

This enzyme-treated RJ is obtained by subjecting RJ to a proteolytic enzyme treatment using an endo-type neutral protease derived from Bacillus subtilis.
RJ is a milky white jelly-like substance made by mixing secretions secreted from the hypopharyngeal gland and the greater vagina by bees aged 3 to 12 days. This RJ is known to have a favorable physiological activity on the human body. The main physiologically active components in this RJ include, for example, proteins, saccharides, organic acids such as 10-hydroxydecenoic acid specific to RJ, lipids, vitamin Bs, vitamins such as folic acid, nicotinic acid and pantothenic acid, Various minerals etc. are mentioned. As RJ used for the drink of this embodiment, either raw RJ or RJ powder obtained by drying and pulverizing raw RJ may be employed. The production area of RJ may be any of China, Brazil, European countries, Oceania countries, the United States, and the like.

  The proteolytic enzyme treatment is performed by hydrolyzing the peptide bond of the protein contained in RJ using an endo-type neutral protease derived from Bacillus subtilis to produce Ile-Tyr, Val-Tyr, Ile- This refers to a treatment for obtaining a peptide such as Val-Tyr. The endo-type neutral protease derived from Bacillus subtilis belongs to the metalloproteinase, and the optimum pH is generally in the range of 6.5 to 7.5.

  This method for producing an enzyme-treated RJ includes a decomposition step, an enzyme deactivation step, and a drying step. The decomposition step is a step of subjecting RJ to a proteolytic enzyme treatment using Bacillus subtilis-derived endo-type neutral protease. The enzyme deactivation step is a step of deactivating the endo-type neutral protease used in the decomposition step. The drying step is a step of drying RJ after the enzyme deactivation step.

  Each step is described below. In the decomposition step, it is preferable to use an RJ diluted solution obtained by diluting the above RJ with a predetermined amount of water or a buffer solution in order to reduce the viscosity of the solution and smoothly advance the enzyme reaction (decomposition reaction). Further, at this time, it is preferable to adjust the pH of the RJ diluted solution to be close to the optimum pH of the endo-type neutral protease. Specifically, the pH of the RJ diluted solution is preferably 6.5 to 7.5, particularly It is preferable to adjust to 6.8 to 7.2. In preparing the RJ dilution, in order to efficiently proceed with the proteolytic enzyme treatment and to rapidly proceed with the drying step, the amount is preferably 2 to 10 times the weight of RJ, more preferably 2. It may be diluted with 5 to 6 times the amount of water or buffer.

  In addition, since RJ contains many components and may inhibit the endo-type neutral protease, it is 10 times the amount when enzyme treatment is performed on a normal substrate. It is preferable to use the above. However, this can also be arbitrarily selected in view of the reaction time. This proteolytic enzyme treatment is preferably performed at 40 to 60 ° C., which is the optimum temperature for the endo-type neutral protease.

  The enzyme deactivation step is a step of heating the RJ diluted solution after the decomposition step to a temperature at which the endo-type neutral protease is deactivated. The heating temperature in this enzyme deactivation step is preferably 75 to 100 ° C. in order to sufficiently deactivate the endo-type neutral protease. The heating time is preferably 5 to 60 minutes. On the other hand, a drying process should just be performed according to a conventional method, for example, is carried out by freeze-drying the RJ dilution liquid after the said enzyme deactivation process using a vacuum freeze dryer etc. By performing this drying step, a powdery enzyme-treated RJ is obtained.

  Ile-Tyr, Val-Tyr, and Ile-Val-Tyr obtained by proteolytic enzyme treatment of RJ each have ACE inhibitory activity and aldosterone secretion inhibitory activity. Furthermore, high ACE inhibitory activity and aldosterone secretion inhibitory activity are also present in peptides consisting of the amino acid sequences represented by SEQ ID NOs: 1 to 3 obtained by the proteolytic enzyme treatment and other unidentified components.

  This drink is, for example, configured to have a volume of about 100 ml because it is easily taken orally, and the solid content of the enzyme-treated RJ is 600 in order to exert a blood pressure lowering effect in the 100 ml. About ~ 1200mg is contained. At this time, the content of the enzyme-treated RJ (solid content) in the drink is 0.6 to 1.2% by mass, and as it is, it is difficult to disperse in water, and insoluble matters such as proteins and peptides precipitate. Or it becomes easy to aggregate.

  Gellan gum is a natural polymeric polysaccharide composed of sugars such as D-glucose, D-glucuronic acid, and D-rhamnose. This gellan gum is contained in order to impart viscosity to the drink, and is contained in order to suppress precipitation and aggregation of the insoluble matter. The content of gellan gum in the drink is preferably from 0.01 to 0.09 mass%, more preferably from 0.02 to 0.05 mass%. When the gellan gum content is less than 0.01% by mass, the drink is not sufficiently viscous, and when it exceeds 0.09% by mass, it is uneconomical.

The drink of this embodiment may contain various additives such as sugar solution, acidulant, vitamins other than vitamin C, fragrance, and pigment.
In this embodiment, fructose glucose liquid sugar and honey are preferably used as the sugar liquid. Fructose-glucose liquid sugar is a sugar liquid produced by starch degradation, isomerization reaction, etc., and has a fructose content higher than that of glucose (for example, fructose is 55% by mass). This fructose-glucose liquid sugar is contained as a sweetener that imparts sweetness to the drink. The content of fructose-glucose liquid sugar in the drink is preferably 3-20% by mass, more preferably 4-15% by mass. When the content of fructose-glucose liquid sugar is less than 3% by mass, the sweetness is low and there is a high possibility that a good flavor is not sufficiently imparted. On the other hand, when it exceeds 20 mass%, excessive sweetness will be provided to a drink and there exists a high possibility that a flavor will fall.

  Honey is contained as a sweetener that imparts sweetness to the drink, similar to the fructose-glucose liquid sugar. The content of honey in the drink is preferably 7% by mass or less. When the content of honey exceeds 7% by mass, excessive sweetness is imparted to the drink, and the possibility of a decrease in flavor is high.

  The sour agent is used to adjust the pH of the drink to a suitable range (3.5 to 6.0). In this embodiment, citric acid is suitably used as the sour agent. The content of citric acid in the drink is preferably 0.1 to 0.5% by mass, more preferably 0.2 to 0.4% by mass. When the citric acid content is less than 0.1% by mass and the pH of the drink exceeds 6.0, the acidity of the drink becomes poor, and the flavor is likely to be impaired. On the other hand, when the citric acid content exceeds 0.5% by mass and when the pH of the drink is less than 3.5, there is a high possibility that an excessive bitter taste will be imparted to the drink. There is also a high possibility of reducing the stability of gellan gum.

  In this embodiment, vitamin B2 is suitably used as a vitamin group other than vitamin C. Vitamin B2 is used to impart nutritional value to the drink. Examples of vitamin B2 include flavin adenine dinucleotide sodium, riboflavin, sodium riboflavin phosphate, riboflavin butyrate, and derivatives thereof. These may be used alone or in combination of two or more. The content of vitamin B2 in the drink is preferably 0.0001 to 0.01% by mass, and more preferably 0.0004 to 0.0008% by mass. When the content of vitamin B2 is less than 0.0001% by mass, there is a high possibility that sufficient nutritional value is not given to the drink. On the other hand, when the content of vitamin B2 exceeds 0.01% by mass, vitamin B2 is excessively consumed and the flavor of the drink is likely to be impaired.

  A fragrance | flavor is used in order to suppress the bitterness and irritation taste of a drink. This perfume may be any conventionally known perfume, but it is particularly preferable to use an emulsified perfume. The emulsified fragrance is generally called a cloudy and is a fragrance obtained by emulsifying an oil-soluble fragrance using an emulsifier and a stabilizer. The said fragrance | flavor may be used independently and may be used in combination of 2 or more type.

  Moreover, a pigment | dye is used in order to provide favorable palatability to a drink. Examples of the dye include anthocyanin dyes, azaphyrone dyes, carotenoid dyes, betacyanine dyes, and polyphenol dyes. These may be used alone or in combination of two or more.

The drink is filled into bottles, cans or paper packs to become the final product. In addition, at this time, it heat-sterilizes by a conventional method.
In the enzyme-treated RJ, active ingredients such as Ile-Tyr, Val-Tyr, and Ile-Val-Tyr act so as to inhibit the signal transmission of the RAS-aldosterone system. When this enzyme-treated RJ is orally administered, the active ingredient is absorbed from the digester and transferred to the blood, and inhibits ACE activity in specific organs such as the abdominal aorta and lungs. At this time, the active ingredient inhibits the production of angiotensin II from angiotensin I by inhibiting ACE activity in the specific organ. This suppression of angiotensin II production inhibits aldosterone secretion from the adrenal gland layer and suppresses sodium reabsorption from the distal renal tubule. As a result, the circulating plasma volume decreases and the blood pressure decreases. Therefore, according to the enzyme treatment RJ, an increase in blood pressure is suitably suppressed, and hypertension can be prevented and improved.

The effects exhibited by the above embodiment will be described below.
-Since the drink preparation of this embodiment contains gellan gum, suitable viscosity is provided to the drink preparation. As a result, the various additives, including proteins and peptides contained in the enzyme-treated RJ, are stably and uniformly dispersed. That is, precipitation and aggregation of various components (protein, peptide, various additives) are suppressed even when the drink is sterilized by heating or stored for a long period of time. Therefore, according to this embodiment, it is possible to obtain a high-quality drink agent in which precipitation and aggregation of insoluble matter are suppressed.

  In addition, the drink of this embodiment does not contain vitamin C that reacts with the enzyme-treated RJ protein (the amino group of the protein) and changes into a substance that easily browns. For this reason, even when the drink is sterilized by heating or stored for a long period of time, browning does not occur and a decrease in flavor is suppressed. That is, according to the drink of this embodiment, good storage stability can be obtained and the quality can be improved.

  ・ By using fructose-glucose liquid sugar, browning is less likely to occur. Glucose is easy to react with vitamin C to cause browning (Maillard reaction), and fructose-glucose liquid sugar having a high content of fructose is excellent in preventing browning. That is, according to the drink of this embodiment, the browning of the drink can be suppressed synergistically by the effect of fructose-glucose liquid sugar.

(Preparation of enzyme-treated RJ)
45 g of water was added to 8 g of RJ produced in China (32.7% solids) and stirred for 5 minutes to prepare an RJ dilution. Next, 25 mg of an endo-type neutral protease derived from Bacillus subtilis (Protein N manufactured by Amano Enzyme Co., Ltd.) was added to the RJ dilution, and the pH was adjusted to 7.0, which was an optimum pH. Reacted for hours. After completion of the reaction, the enzyme inactivation step was performed by heating at 98 ° C. for 5 minutes, and then the powdery enzyme-treated RJ was prepared by performing the drying step by lyophilization. Although not shown, this enzyme-treated RJ contained a peptide consisting of the amino acid sequence represented by SEQ ID NOs: 1 to 3 in addition to Ile-Tyr, Val-Tyr and Ile-Val-Tyr. Was confirmed by HPLC analysis.

(Examination of emulsifier)
Here, considering the possibility that the insoluble matter generated during dissolution of the enzyme-treated RJ is an oily substance, the properties of the enzyme-treated RJ in which various emulsifiers are added and the insoluble matter is dispersed (whether a precipitate is generated) The emulsifier was screened by visual evaluation. That is, 600 mg of the above enzyme-treated RJ was mixed with a sugar solution, a sour agent, and an emulsifier shown in Table 1 below (the content (w / v%) in the table is the final concentration) to make a total amount of 100 ml with water. Was prepared and then heat sterilized at 75 ° C. for 20 minutes. Properties of the enzyme-treated RJ at the time of mixing and immediately after heat sterilization were evaluated and are shown in Table 1. In Table 1, the HLB (Hydrophilic-Lipophilic Balance) value of each emulsifier is also shown.

表１から明らかなように、乳化剤の添加のみでは、沈殿（不溶解物）を分散させることはできなかった。なお、これら乳化剤の中でも、ショ糖ラウリン酸エステル、ラウリン酸デカグリセリン等のラウリン酸系乳化剤に関しては、混合時には一旦均一となって比較的性状は良かったが、経時的に沈殿が発生した。

As is apparent from Table 1, the precipitate (insoluble matter) could not be dispersed only by the addition of the emulsifier. Among these emulsifiers, lauric acid-based emulsifiers such as sucrose laurate and decaglycerin laurate were once uniform and relatively good in properties, but precipitation occurred over time.

(Examination of various additives for enzyme-treated RJ)
An enzyme-treated RJ (600 mg) and other additives shown below were mixed to prepare a mixed solution having a total volume of 100 ml with water, and the influence of these additives was examined. Examples of the additive include sugar solution of oligosaccharide (10%, 23.5%), sugar (9%, 18%), maltitol (15%, 30%), erythritol (5%), and SE (SE). (10, 25%), the emulsifier was fixed to Q12S, the thickening polysaccharide was fixed to xanthan gum 0.1%, and vitamin C concentrations were added to 0, 0.6, and 1.2%, respectively. In addition, the number in the said parenthesis shows the percentage of the mass of each additive with respect to the volume of a liquid mixture. Then, with respect to the various mixed liquids, changes with time in properties were visually evaluated under refrigeration, room temperature, 40 ° C., and 80 ° C., respectively. Among these mixed solutions, typical formulation examples are shown in Table 2 below. Moreover, Bx and pH of various liquid mixtures are shown together in the same table.

各混合液について上記温度条件でそれぞれ加速試験を行い、その後、各混合液の性状を確認するとともに、ＨＰＬＣを用いてそれぞれのペプチド減少量を評価した。その結果、いずれの混合液に関しても、性状及びペプチド減少量についての大きな変化はみられなかった。乳化剤の有無に関しても、性状及びペプチド減少量について大きな変化はみられなかった。また、Ｂｘの値が高い混合液ほど、性状が安定していた。加えて、各混合液を４０℃で加速保存すると、それぞれ褐変化が進行することが確認された。

Each mixed solution was subjected to an acceleration test under the above temperature conditions, and then the properties of each mixed solution were confirmed, and each peptide decrease amount was evaluated using HPLC. As a result, no significant change was observed in the properties and peptide loss for any of the mixed solutions. There was no significant change in properties and peptide loss with or without emulsifiers. In addition, the higher the Bx value, the more stable the properties. In addition, when each liquid mixture was accelerated and stored at 40 ° C., it was confirmed that browning proceeded.

(Examination of browning suppression)
Next, screening was performed for substances that cause browning (browning substances) from basic additives formulated in general drinks. In addition, in the liquid mixture which mixed enzyme treatment RJ with water, since not only the appearance property but the stability of a peptide should be evaluated, the stability of the peptide was also examined here. Specifically, the following sugar solution screening, examination of vitamin C concentration, examination of pH, and examination of the relationship between the content of enzyme-treated RJ and vitamin C were performed.

<Screening of sugar solution>
Here, the effect of various sugar solutions on the browning of the drink was examined. As a model solution, three kinds of peptide standard products (VY + IY + IVY: 0.5 mg + 0.5 mg + 1.2 mg) which are active ingredients, citric acid and vitamin C were mixed to prepare a mixed solution with a total volume of 100 ml with water. Various sugar solutions shown in Table 3 below were added to these mixed solutions, and an accelerated test was conducted at 80 ° C. for 3 days. Then, HPLC analysis is performed on the mixed solution (room temperature, 0 day) containing isomaltoligosaccharide, and the remaining peptide residues in the various mixed solutions are determined from the peak area (AREA) based on the peak of each peptide obtained. The rate was calculated. Furthermore, the absorbance (OD420) in the brown region (420 nm) was measured for each mixed solution after completion of the acceleration test. The results are shown in Table 3.

表３から明らかなように、各種混合液の褐変化とペプチドの減少との間には、特に相関関係はみられなかった。また、各種混合液中のペプチドに関しては、ＶＹ及びＩＹの減少が著しく、ＩＶＹは比較的安定していることが確認された。

As is clear from Table 3, no particular correlation was observed between the browning of the various liquid mixtures and the decrease in peptides. Moreover, regarding the peptides in various mixed solutions, the decrease in VY and IY was remarkable, and it was confirmed that IVY was relatively stable.

<Examination of vitamin C concentration>
Here, the browning change and pH when the vitamin C concentration was appropriately changed were examined. That is, as a model solution, three kinds of peptide standard products (VY + IY + IVY: 0.5 mg + 0.5 mg + 1.2 mg), various sugar solutions (25 g), peptides (VY + IY + IVY: 0.5 mg + 0.5 mg + 1.2 mg), citric acid (0.4 g) and ascorbic acid (0 to 0.4 g) were mixed to prepare a mixed solution having a total volume of 100 ml with water. These mixed solutions were subjected to an acceleration test at 80 ° C. for 3 days. And the pH of the liquid mixture after completion | finish of an acceleration test, and the light absorbency (OD420) in a brown region (420 nm) were measured. The results are shown in Table 4.

表４に示すように、いずれの糖液を用いた場合でも、アスコルビン酸の濃度が増加するに従って、加速試験後の混合液の褐変化が顕著になることが確認された。このため、アスコルビン酸をドリンク剤の処方から抜くことで、ドリンク剤の褐変化はかなり抑制されるものと思われる。

As shown in Table 4, it was confirmed that even when any sugar solution was used, the browning of the mixed solution after the acceleration test became remarkable as the concentration of ascorbic acid increased. For this reason, it is considered that the browning of the drink is considerably suppressed by removing ascorbic acid from the drink formulation.

<Examination of pH>
Here, the correlation between pH and browning was examined. That is, as a model solution, three kinds of peptide standard products (VY + IY + IVY: 0.5 mg + 0.5 mg + 1.2 mg), various sugar solutions (25 g), citric acid (0.28 to 0.4 g), citric acid 3Na (0-0.12g) and ascorbic acid (0.2g) were mixed and the liquid mixture made into 100 ml in total with water was prepared. These mixed solutions were subjected to an acceleration test at 80 ° C. for 3 days. And the pH of the liquid mixture after completion | finish of an acceleration test, and the light absorbency (OD420) in a brown region (420 nm) were measured. The results are shown in Table 5. At this time, the mixture was prepared such that the substitution rate of citric acid 3Na was 0, 10, 20, and 30%, and the total weight of citric acid and citric acid 3Na was 0.4 g.

表５に示すように、いずれの糖液を用いた場合でも、クエン酸３Ｎａの置換率を増加させて混合液のｐＨを上げることにより褐変化が若干抑制された。これは、ｐＨが高いほど、アスコルビン酸の酸化が抑制され、その結果、混合液の褐変化が抑制されたものと推測される。

As shown in Table 5, even when any sugar solution was used, the browning was slightly suppressed by increasing the substitution rate of 3Na citrate to raise the pH of the mixed solution. This is presumed that as the pH is higher, the oxidation of ascorbic acid is suppressed, and as a result, the browning of the mixed solution is suppressed.

<Examination of the relationship between the content of enzyme-treated RJ and vitamin C>
Here, it was examined whether or not the content of the enzyme-treated RJ was appropriately changed to influence the browning. That is, a certain amount of vitamin C was mixed with a mixed solution obtained by mixing 250 mg, 500 mg or 600 mg of the enzyme-treated RJ in 100 ml of water. And after performing the acceleration test for 60 days at 40 degreeC about each liquid mixture, the brown change was evaluated by the light absorbency in 420 nm. As a result, data is not shown, but as the content of the enzyme-treated RJ increases, the browning of the mixture becomes significant. This is presumed that the greater the content of the enzyme-treated RJ, the more amino groups that contribute to the reaction with vitamin C, and thus the more easily the browning proceeds.

  As a result, it was found that the main factor of browning of the mixed solution containing the enzyme-treated RJ is vitamin C, and the content of the enzyme-treated RJ is also involved in the browning. On the other hand, regarding the sugar solution, the effect on the browning was slight. Therefore, in the following test, vitamin C is excluded from the drink formulation.

(Examination of thickening polysaccharides)
Here, a thickened polysaccharide was screened with respect to a mixture obtained by mixing the enzyme-treated RJ in 100 ml of water in order to disperse and stabilize insoluble matters (proteins and peptides) in the mixture. That is, enzyme-treated RJ 600 mg or 1200 mg (0.6 to 1.2% by mass), various sugar solutions (13.5% by mass) shown in Table 6 below, thickening polysaccharide (0.03 to 2.2% by mass) ), Vitamin C (0.2% by mass), acidulant (0.45% by mass) and flavor (0.19% by mass) were added to 100 ml of purified water to prepare a mixed solution. And various liquid mixture was heat-sterilized (75 degreeC, 20 minutes), and the property was evaluated about each liquid mixture after performing an accelerated test (40 degreeC, 60 days). The results are shown in Table 6.

表６から明らかなように、ペクチンを含有する混合液は、加熱殺菌後に沈殿が生じた。一方、その他の増粘多糖類を含有する混合液は、経時的に沈殿が生じ、比較的安定な性状が得られた。これらの中でも、増粘多糖類として０．０３質量％のジェランガムを含有する混合液は、加速試験後にも沈殿が認められず、最も良好な性状となっていた。すなわち、これらの増粘多糖類の中でも、ジェランガムが、不溶解物の生成抑制に特に優れていることが確認された。

As is clear from Table 6, precipitation occurred in the mixed solution containing pectin after heat sterilization. On the other hand, the mixed liquid containing other thickening polysaccharides was precipitated over time, and a relatively stable property was obtained. Among these, the liquid mixture containing 0.03% by mass of gellan gum as the thickening polysaccharide showed no precipitation even after the acceleration test and had the best properties. That is, among these thickening polysaccharides, it was confirmed that gellan gum is particularly excellent in suppressing the formation of insoluble matter.

(Screening of sugar solution without vitamin C)
Here, vitamin C was excluded from the drink formulation, and the sugar solution was screened in the presence of the enzyme-treated RJ. That is, 600 mg of enzyme-treated RJ, 20 g of various sugar solutions shown below, 0.45 g of citric acid and 0.03 of gellan gum were mixed to prepare a mixed solution with a total volume of 100 ml with water. After storing these mixed solutions at 40 ° C. for 60 days, each browning change was evaluated by absorbance at OD420. The results are shown in Table 7.

表７から明らかなように、ドリンク剤の処方からビタミンＣを除外することで、ドリンク剤の褐変化が好適に抑制された。また、糖液の種類における褐変化の差異は特にみられなかった。

As apparent from Table 7, the browning of the drink was suitably suppressed by excluding vitamin C from the drink formulation. Moreover, the difference of the brown change in the kind of sugar liquid was not seen especially.

  Here, vitamin C was excluded from the formulation, and various drinks shown in Table 8 below were added to a mixed solution obtained by mixing 1200 mg of the enzyme-treated RJ in 100 ml of water to prepare a trial drink. In addition, the number shown in Table 8 shows the percentage of the mass of each component with respect to the volume of a drink. That is, the content (stability) of the drink was evaluated by fixing the enzyme-treated RJ content in the drink to 1200 mg, the gellan gum content to 0.03%, and appropriately changing Bx. The results are shown in Table 8.

表８の結果より、各種ドリンク剤において、ジェランガムの増粘性が好適に発揮されて、いずれの糖液含量でも沈殿を生じることなく良好な性状（安定性）が得られた。従って、表８に示す成分の組み合わせにより、特に優れた性状を有するドリンク剤が得られる。また、加速試験時の安定性、最終製品のカロリー及び製造コストの点からも、表８に示す処方例が優れていることが確認された。

From the results of Table 8, in various drinks, the thickening of gellan gum was suitably demonstrated, and good properties (stability) were obtained without causing precipitation at any sugar solution content. Therefore, a drink having particularly excellent properties can be obtained by combining the components shown in Table 8. Moreover, it was confirmed that the formulation example shown in Table 8 is excellent also from the viewpoint of stability during the acceleration test, calories of the final product, and production cost.

In addition, this embodiment can also be changed and embodied as follows.
-You may add a conventionally well-known crude drug, herb extract, a vegetable fiber, minerals, a preservative, etc. to a drink agent in combination as needed suitably.

  -Vitamin C may be included in the drink. In this case, although the effect regarding suppression of the brown change of a drink agent falls, since the said vitamin C is not concerned at all about precipitation of an insoluble matter (protein, peptide), it suppresses precipitation of an insoluble matter. The object of the present invention is achieved in the same manner as the present embodiment.

Further, the technical idea that can be grasped from the embodiment will be described below.
The enzyme-treated royal jelly-containing drink according to claim 2, wherein the sugar liquid contains fructose-glucose liquid sugar. In this case, since the fructose-glucose liquid sugar has a high content of fructose that hardly causes browning, the browning of the drink can be further suppressed.

Claims (4)

An enzyme-treated royal jelly-containing drink comprising an enzyme-treated royal jelly that has been subjected to proteolytic enzyme treatment using an endo-type neutral protease derived from Bacillus subtilis, and gellan gum. Enzyme-treated royal jelly, which comprises an enzyme-treated royal jelly treated with an endo-type neutral protease derived from Bacillus subtilis, gellan gum, sugar solution, acidulant, vitamins other than vitamin C, fragrance, pigment, and water Contains drinks. The enzyme-treated royal jelly is contained according to claim 1 or 2, wherein the enzyme-treated royal jelly is contained in an amount of 0.6 to 1.2% by mass, and the gellan gum is contained in an amount of 0.01 to 0.09% by mass. Drink agent. The enzyme-treated royal jelly-containing drink according to any one of claims 1 to 3, which has a blood pressure lowering effect.