JP5248755B2 - Angiotensin I converting enzyme inhibitor and method for producing the same - Google Patents

Description

The present invention is a cacao nib or cacao jam and / or an extract of cacao nib or cacao jam with water, methanol, ethanol, ethyl acetate, or a mixed solvent thereof selected from the group consisting of polyvinylpyrrolidone, methacrylate polymer and polyphenol oxidase The present invention relates to an angiotensin I converting enzyme inhibitor and a method for producing the same.

  Hypertension is a cause of arteriosclerosis along with diabetes, hyperlipidemia, obesity and the like. Moreover, since serious complications may accompany it, much attention has been given to its treatment and prevention.

  As one of the solutions for hypertension, it is known that inhibiting angiotensin I converting enzyme (hereinafter abbreviated as ACE) in the renin-angiotensin system, which is known as a mechanism for raising blood pressure, is effective. ACE exists mainly in lungs, vascular endothelial cells, and renal proximal tubules, and is an enzyme that converts angiotensin I, which is a substrate, into angiotensin II. This angiotensin II acts as a powerful blood pressure increasing hormone and promotes vasoconstriction by binding to receptors present in blood vessels. It is also known that ACE has an activity of degrading kinin-kallikrein (hypertensive) bradykinin. Since inhibition of ACE activity suppresses production of angiotensin II and degradation of bradykinin, ACE inhibitors are clinically effective for the treatment and prevention of hypertension.

  Recently, it has been found that ingredients derived from food and natural products have ACE inhibitory activity. For example, casein (see, for example, Patent Document 1 and Non-Patent Document 1), sardine (for example, see Patent Document 2 and Non-Patent Document 2), seaweed (see, for example, Patent Documents 3, 4, 5, 6, and 7), Peptides obtained from incubation eggs (for example, see Patent Document 8), polyphenols such as tea polyphenol (for example, see Non-Patent Document 3), fruit polyphenol (for example, see Patent Document 9), and other eucalyptus extracts ( For example, plant origin extracts, such as a soybean extract (for example, refer patent document 11), etc. are mentioned.

  On the other hand, cacao (Theobroma cacao) also has a blood pressure lowering effect, and it reduces the loss of polyphenols by weakening the degree of fermentation in the production process, thereby reducing blood pressure activity in cocoa extracts and the like with an increased amount of cacao polyphenols. (For example, see Patent Documents 12 and 13) and reports that lignin, which is a kind of polyphenol contained in cacao, decreased blood pressure in spontaneously hypertensive rats (for example, Non-Patent Document 4). reference). Further, a composition containing a cholesterol-lowering agent and cocoa polyphenol (for example, see Patent Document 14) or a composition containing L-arginine and cocoa polyphenol (for example, see Patent Document 15) has a therapeutic and preventive effect on heart disease. It has been made clear. However, it is not clear that components other than cocoa polyphenols show ACE inhibitory activity, and that cocoa-derived amino acid compositions show ACE inhibitory activity.

JP-A-6-128287 JP 7-188282 A JP 2002-138100 A Japanese Patent Laid-Open No. 2003-246795 JP 2003-246696 A JP 2003-246797 A JP 2003-252897 A JP 2005-145827 A Japanese Patent Laid-Open No. 2002-47196 Japanese Patent Laid-Open No. 11-60498 JP-A-2005-95156 JP 2003-204758 A JP 2001-500016 A JP 2003-530410 A JP 2002-505864 A Bioscience Biotechnology and Biochemistry, 58, 12, 2244-2245, 1994 Japanese Journal of Nutrition and Food Science, 53, 2, 77-85, 2000 Japanese Journal of Agricultural Chemistry, 61, 803-808, 1987 Japanese Journal of Agricultural Chemistry, 68, 957-965, 1994

  The inventors of the present invention performed depolyphenol treatment on cocoa beans and / or cocoa bean extracts, and found that the obtained treated products have high ACE inhibitory activity. Further, the present inventors have found that an amino acid composition derived from cocoa beans and / or cocoa bean extracts has a high ACE inhibitory activity. As described above, there are many reports on the blood pressure lowering effect of cocoa, but polyphenols contained in cocoa are only shown as effective ingredients, and components other than polyphenols in cocoa show ACE inhibitory activity This is the first time that the present invention has been revealed.

  That is, an object of the present invention is to provide a natural product-derived ACE activity inhibitor that is safe and highly active as a food, a method for producing the same, and a food or drink containing the same.

  In order to solve the above-mentioned problems, the present inventors performed depolyphenol treatment on cocoa beans and / or cocoa bean extracts, and conducted an ACE activity inhibition test. Specifically, a cocoa bean extract obtained by extracting cocoa beans and / or cocoa beans that have not been extracted using a solvent such as water, methanol, ethanol, acetone, ethyl acetate, or a mixed solvent thereof. On the other hand, a fraction containing a high amount of amino acids by removing polyphenols efficiently by performing treatment with an adsorption aid such as polyvinylpyrrolidone or enzyme treatment and further concentrating the amino acid composition such as column fractionation. And found that these fractions have significant ACE inhibitory activity, thus completing the present invention.

That is, the present invention is a group comprising an extract of cacao nibs or cacao jams and / or cacao nibs or cacao jams with water, methanol, ethanol, ethyl acetate, or a mixed solvent thereof, polyvinylpyrrolidone, methacrylate polymer and polyphenol oxidase. More particularly, the present invention relates to an angiotensin I converting enzyme inhibitor that is subjected to at least one depolyphenol treatment selected from the above, and a method for producing the same.

The present invention relates to an angiotensin I-converting enzyme inhibitor comprising further subjecting the depolyphenol-treated product to an amino acid composition concentration treatment by column fractionation, and a method for producing the same.

  As described above, the angiotensin I converting enzyme inhibitor of the present invention has an action of effectively inhibiting ACE involved in an increase in blood pressure. Therefore, by containing the angiotensin I converting enzyme inhibitor of the present invention in various foods and drinks, it can be used for the treatment or prevention of hypertension. In addition, the angiotensin I converting enzyme inhibitor of this invention has a remarkably high ACE inhibitory effect compared with the sadden peptide made effective in the blood pressure fall.

  Since the angiotensin I converting enzyme inhibitor of the present invention is derived from food (cocoa beans), there is no problem in safety.

  In addition, since the angiotensin I converting enzyme inhibitor of the present invention has been subjected to depolyphenol treatment, it is expected to have an effect that it has less bitterness peculiar to polyphenols and is excellent in taste and can be ingested more easily than before. it can.

  The product of the present invention can be applied to various foods and beverages and preparations. Moreover, all the cocoa beans used as the raw material of the product of the present invention have been used as food and drink materials and natural additives for a long time, and there is no problem with regard to their safety.

  The present invention is described in detail below.

  As the cocoa beans that are the raw materials of the present invention, cocoa nibs (edible part) or cocoa jam (embryo part) that has been heat-treated are usually used, and the heat-treatment method and parts thereof are not limited thereto.

  Although it does not specifically limit about the method of obtaining a cocoa bean extract from cocoa bean, 1 type, or 2 or more types of mixed solvents of organic solvents, such as water, lower alcohols, such as methanol, ethanol, and n-propanol, and ethyl acetate, The cocoa bean extract of the present invention can be obtained by a conventional extraction method. However, considering that the product of the present invention is used by humans as foods and beverages or preparations, it is preferable to use a combination of water and ethanol as the extraction solvent from the viewpoint of safety.

  As extraction conditions, extraction can be performed at any temperature of 20 to 90 ° C, but 50 to 80 ° C for about 1 to 5 hours is preferable. The obtained extract can be used after being filtered and further concentrated or lyophilized under reduced pressure.

  Examples of the treatment for removing polyphenol from the cocoa bean extract or cocoa bean itself include solvent extraction treatment such as water acetone, adsorption aid treatment such as Sephadex, methacrylate polymer, polyvinyl polypyrrolidone, polyphenol oxidase, etc. Enzyme treatment, ferric chloride method, and combinations of these methods.

  Furthermore, as the amino acid composition concentration treatment, a highly amino acid-containing composition can be obtained by purifying the treated product using an organic solvent, ion exchange column chromatography or the like such as column fractionation. This high amino acid-containing composition is considered to consist of protein degradation products, peptides, and amino acids.

  In addition, since the angiotensin I converting enzyme inhibitor of the present invention is excellent in aroma, taste and safety, for example, foods such as chewing gum, candy, tablet confectionery, gummy jelly, chocolate and other confectionery, sherbet, beverage In addition, it can be blended into preparations such as tablets and gargles and used on a daily basis.

  The added amount is about 0.001% by weight or more, preferably about 0.01% by weight or more in the state of a dried product with respect to the food or drink or the preparation.

  The cocoa beans used as the raw material of the present invention have been used for a long time as food materials and natural additives, and there is no problem with the safety of these processed products, foods and drinks and formulations containing them. .

  EXAMPLES Hereinafter, although a test example is given and this invention is demonstrated in detail, they do not restrict | limit the range of this invention goods.

Test example 1

  This test was conducted to prepare an extract from cocoa beans.

1) Test sample Cacao nibs and cacao jars were used.

2) Test method An extract was prepared as follows.

i) Solvent extraction of cocoa nibs 1.5 L of 50% ethanol was added to 200 g of cocoa nibs, and the mixture was allowed to stand at 80 ° C. for 1 hour for extraction. This was filtered, and the filtrate was concentrated under reduced pressure and then lyophilized. This operation was repeated twice to obtain 25 g of the dried extract NA.

ii) Solvent extraction of cocoa nibs 1.5 L of water was added to 200 g of cocoa nibs, and the mixture was allowed to stand at 70 ° C. for 3 hours for extraction. This was filtered, and the filtrate was concentrated under reduced pressure and then lyophilized. By repeating this operation twice, 30 g of the dried extract NB was obtained.

iii) Solvent extraction of cacao nibs 1 L of 100% ethanol was added to 200 g of cacao nibs, and extraction was performed by allowing to stand at 60 ° C. for 1 hour. This was filtered, and the filtrate was dried under reduced pressure. By repeating this operation twice, 10 g of the dried extract NC was obtained.

iv) Solvent extraction of cocoa nibs 1 L of 100% methanol was added to 200 g of cocoa nibs, and the mixture was allowed to stand at 60 ° C. for 2 hours for extraction. This was filtered, and the filtrate was dried under reduced pressure. By repeating this operation twice, 11 g of an extract dried product ND was obtained.

v) Solvent extraction of cocoa nibs 1 L of 100% ethyl acetate was added to 200 g of cocoa nibs, and the mixture was allowed to stand at 70 ° C. for 1 hour for extraction. This was filtered, and the filtrate was dried under reduced pressure. By repeating this operation twice, 10 g of the dried extract NE was obtained.

vi) Solvent extraction of cocoa jam 600 mL of 50% ethanol was added to 90 g of cocoa jam, and the mixture was allowed to stand at 50 ° C. for 3 hours for extraction. This was filtered, and the filtrate was concentrated under reduced pressure and then lyophilized. This operation was repeated twice to obtain 12 g of the dried extract JA.

vii) Solvent extraction of cacao jam 1.2 L of water was added to 180 g of cacao jam, and the mixture was allowed to stand at 70 ° C. for 3 hours for extraction. This was filtered, and the filtrate was concentrated under reduced pressure and then lyophilized. This operation was repeated twice to obtain 12 g of the dried extract JB.

viii) Solvent extraction of cacao jam 1.5 L of 100% ethanol was added to 200 g of cacao jam, and the mixture was allowed to stand at 60 ° C for 1 hour for extraction. This was filtered, and the filtrate was dried under reduced pressure. By repeating this operation twice, 10 g of the dried extract JC was obtained.

ix) Solvent extraction of cacao jam 1.2 L of 100% methanol was added to 180 g of cacao jam, and the mixture was allowed to stand at 60 ° C for 2 hours for extraction. This was filtered, and the filtrate was dried under reduced pressure. By repeating this operation twice, 12 g of dried extract JD was obtained.

x) Solvent extraction of cocoa jam 1.5 L of 100% ethyl acetate was added to 200 g of cocoa jam, and the mixture was allowed to stand at 70 ° C. for 1 hour for extraction. This was filtered, and the filtrate was dried under reduced pressure. By repeating this operation twice, 12.5 g of the dried extract JE was obtained.

3) Test results Extracts NA to NE were obtained from cacao mass by this treatment. In addition, extracts JA to JE were obtained from cacao jam.

Test example 2

  This test was conducted to prepare a depolyphenol-treated product from the cocoa bean extract.

1) Test sample Each solvent extract (NA to NE) of cacao nibs prepared in Test Example 1 and each solvent extract (JA to JE) of cacao jam were used.

2) Test method 10 g of each extract was dissolved in 500 mL of the same solvent as the extraction solvent, and 50 g of polyvinyl polypyrrolidone (PVPP) as a polyphenol adsorbent was added thereto and stirred and adsorbed for 2 hours to remove polyphenol. Thereafter, the solution was filtered with suction, and the filtrate was concentrated under reduced pressure and freeze-dried.

3) Test results 6.0 g, 5.0 g, 5.0 g, 5.5 g, and 6.0 g of depolyphenol-treated products (NAP to NEP) of each cocoa nib extract (NA to NE) were obtained, respectively. 6.0 g, 5.5 g, 6.0 g, 5.5 g, and 6.0 g of depolyphenol-treated products (JAP to JEP) of each cocoa jam extract (JA to JE) were obtained, respectively.

Test example 3

  This test was conducted to prepare a depolyphenol-treated product from cocoa beans.

1) Test sample Cocoa nibs and cacao germs were used.

2) Test method An extract was prepared as follows.

i) Depolyphenol treatment of cocoa nibs 1.5 L of water was added to 200 g of pulverized cocoa nibs, and 50 g of polyvinyl pyrrolidone (PPVP) as a polyphenol adsorbent was added, followed by stirring and adsorbing for 24 hours to remove polyphenols. Then, it extracted by leaving still at 80 degreeC for 1 hour. This was filtered, and the filtrate was concentrated under reduced pressure and then lyophilized. This operation was repeated twice to obtain 2.5 g of dried extract NP.

ii) Depolyphenol treatment of cocoa jam Add 1.5 L of 50% acetone to 200 g of pulverized cocoa jam, add 50 g of polyvinylpyrrolidone (PPVP), a polyphenol adsorbent, and remove the polyphenol by stirring and adsorbing for 24 hours. did. Then, it extracted by leaving still at 45 degreeC for 1 hour. This was filtered, and the filtrate was concentrated under reduced pressure and then lyophilized. This operation was repeated twice to obtain 3 g of a dried extract JP.

Test example 4

  This test was performed to concentrate the amino acid composition of the depolyphenol-treated product.

1) Test sample NAP, NBP, JAP, and JBP prepared in Test Example 2 were used.

2) Test method i) Treatment with cation exchange resin Column adsorbed fraction and non-adsorbed fraction by Dowex 50W × 4 (The Dow Chemical Company) prepared NAP, NBP, JAP, JBP in H ⁺ foam It was fractionated. That is, 5 g of the test sample was applied to Dowex 50W × 4, eluted with 3 L of water to remove the unadsorbed portion, and then eluted with 2.5 L of 2N aqueous ammonia. The obtained elution fraction was concentrated and freeze-dried.

ii) Treatment by ion exchange gel filtration NAP, NBP, JAP and JBP were fractionated into a column adsorbed fraction and a non-adsorbed fraction by SP Sephadex C-25 (Pharmacia) prepared in H ⁺ form. That is, 5 g of the donated sample was applied to SP Sephadex C-25, eluted with 3 L of water to remove the unadsorbed portion, and then eluted with 2.5 L of 2% NaCl aqueous solution. The fraction was further fractionated by Dowex 50W × 4 (The Dow Chemical Company) and desalted. The obtained elution fraction was concentrated and freeze-dried.

3) Test results Fractions NAPD, NBPD, JAPD, and JBPD in which the amino acid composition was concentrated from NAP, NBP, JAP, and JBP were obtained by treatment with a cation exchange resin. In addition, fractions NAPS, NBPS, JAPS, and JBPS in which the amino acid composition was concentrated from NAP, NBP, JAP, and JBP were obtained by treatment with ion exchange gel filtration.

Test Example 5

  This test was conducted to measure the ACE inhibitory activity, polyphenol content, and amino acid content of each sample.

1) Test sample The samples prepared in Test Examples 1 to 4 were used.

2) Test method i) Measurement of ACE inhibitory activity It was carried out according to the method described in Biochemical Pharmacology 20.1637, 1971.

  A test solution dissolved in 10% dimethyl sulfoxide was prepared so that the test sample had a final concentration of 500 ppm. Further, hippurylhistidiylleucine as a substrate was dissolved in a solution containing 600 mM sodium chloride in 400 mM phosphate buffer (pH 8.5) to prepare a reaction solution having a concentration of 2 mg / ml. Add 0.1 ml of the reaction solution to 0.05 ml of the test solution, shake at 37 ° C. for 3 minutes, add 0.1 ml of ACE prepared to 0.1 U / ml to the mixed solution, and react at 37 ° C. for 1 hour. I let you. After 1 hour, the reaction was stopped by adding 0.5 ml of 1N hydrochloric acid. In order to obtain hypuric acid produced by this reaction, 1.5 ml of ethyl acetate was added to the reaction solution and extracted. Hyprilic acid was quantified by HPLC, and ACE of the test sample was determined according to the following formula (A = peak area of hypuric acid extracted from the test sample, B = peak area of hippuric acid extracted from 10% dimethyl sulfoxide). The inhibition rate was determined.

      ACE inhibition rate (%) = (1−A / B) × 100

ii) Measurement of polyphenol content The content of polyphenol was measured using the Folin-Ciocalteu method.

  To a 50 mL volumetric flask, 0.1 g of a test sample, 35 ml of distilled water, and 5 ml of Folin-Ciocalteu reagent (Sigma) were added, mixed and allowed to stand for 1 minute. Thereafter, 5 ml of a 20% sodium carbonate solution was added and immediately made up to 50 ml. These were mixed and allowed to stand for 1 hour, and then the absorbance at a wavelength of 765 nm was measured. As a sample for the calibration curve, (−)-epicatechin (Sigma) was used, and based on this, the amount of polyphenol of each sample was determined.

iii) Measurement of protein amount The protein amount (amino acid composition amount) was measured using the Kjeldahl method.

  Add 1 g of test sample, 10 ml of sulfuric acid, 8 ml of 30% hydrogen peroxide, and appropriate amount of decomposition accelerator to the Kjeldahl reaction tube, and decompose protein at 420 ° C using a heating device for decomposition (Ticator). went. 100 ml of pure water was added to the decomposition solution, and the nitrogen content was determined with a distillation / titrator (Ticator). The nitrogen content is represented by the following formula.

Nitrogen (%) = 0.014 × (V ₁ −V ₀ ) × f / S × 100
V ₀ : 0.1 N sulfuric acid standard solution volume required for titration (mL): Blank V ₁ : 0.1 N sulfuric acid standard solution volume required for titration (mL): Test sample f: 0.1 N sulfuric acid standard Liquid factor S: Test sample amount (g)

  The protein content was determined from the following formula.

Protein content (%) = Nitrogen (%) * × 6.25 **
*: Value after subtracting caffeine / theobromine nitrogen (%)
**: Protein conversion factor

3) Test results The results are shown in Tables 1 and 2.

  As shown in Tables 1 and 2, the angiotensin I-converting enzyme inhibitor of the present invention obtained by the above method has a composition composed of amino acids as a main component, and by depolyphenol treatment, the amount of amino acid composition is increased and significantly increased. It is an ACE inhibitor showing high activity.

Test Example 6

  This test was conducted in order to examine that the ACE inhibitory activity effect of the amino acid composition of the cocoa bean extract is mainly derived from the amino acid composition.

1) Test Sample NAd prepared by diluting with 10% dimethyl sulfoxide so that the polyphenol content of NA obtained in Test Example 1 is the same value as the polyphenol content of NAP obtained in Test Example 4. NAP was used.

2) Test method ACE inhibition was measured according to the method described in Biochemical Pharmacology 20.1637, 1971.

  A test solution dissolved in 10% dimethyl sulfoxide was prepared so that the test sample had a final concentration of 500 ppm. Further, hippurylhistidiylleucine as a substrate was dissolved in a solution containing 600 mM sodium chloride in 400 mM phosphate buffer (pH 8.5) to prepare a reaction solution having a concentration of 2 mg / ml. Add 0.1 ml of the reaction solution to 0.05 ml of the test solution, shake at 37 ° C. for 3 minutes, add 0.1 ml of ACE prepared to 0.1 U / ml to the mixed solution, and react at 37 ° C. for 1 hour. I let you. After 1 hour, the reaction was stopped by adding 0.5 ml of 1N hydrochloric acid. In order to obtain hypuric acid produced by this reaction, 1.5 ml of ethyl acetate was added to the reaction solution and extracted. Hyprilic acid was quantified by HPLC, and ACE of the test sample was determined according to the following formula (A = peak area of hypuric acid extracted from the test sample, B = peak area of hippuric acid extracted from 10% dimethyl sulfoxide). The inhibition rate was determined.

      ACE inhibition rate (%) = (1−A / B) × 100

3) Test results The results are shown in Table 3.

  As shown in Table 3, the cocoa extract NAP of the present invention showed higher ACE inhibitory activity than NAd despite the polyphenol content being the same. From the above, it was revealed that the amino acid composition of the cocoa extract has a high ACE inhibitory activity.

Test Example 7

  This test is a “marine peptide” (Nisshin Oillio Co., Ltd.) containing the ACE inhibitory activity of the angiotensin I converting enzyme inhibitor of the present invention and a sadden peptide that is one of the foods for specified health use and is effective for lowering blood pressure. This was carried out in order to compare and examine the ACE inhibitory activity of Group Co.).

1) Test sample “Marine peptide” (Nisshin Oilio Group Co., Ltd., protein content 44%), which is one of the foods for specified health use and formulated with a sadden peptide effective for lowering blood pressure, The fraction NAP (a product obtained by depolyphenol treatment of the cocoa nib extract and having a protein content of 30.6%) obtained in the above was used as a test sample.

2) Test method ACE inhibition was measured according to the method described in Biochemical Pharmacology 20.1637, 1971.

  At that time, considering the difference in protein content between the two, the test was conducted after the final concentration of the protein was aligned.

  That is, a solution to be tested was prepared by dissolving 10% dimethyl sulfoxide so that the final concentration of NAP was 500 ppm (marine peptide was 348 ppm). Further, hippurylhistidiylleucine as a substrate was dissolved in a solution containing 600 mM sodium chloride in 400 mM phosphate buffer (pH 8.5) to prepare a reaction solution having a concentration of 2 mg / ml. Add 0.1 ml of the reaction solution to 0.05 ml of the test solution, shake at 37 ° C. for 3 minutes, add 0.1 ml of ACE prepared to 0.1 U / ml to the mixed solution, and react at 37 ° C. for 1 hour. I let you. After 1 hour, the reaction was stopped by adding 0.5 ml of 1N hydrochloric acid. In order to obtain the hypuric acid produced | generated by this reaction, 1.5 ml of ethyl acetate was added and extracted to the reaction solution. Hyprilic acid was quantified by HPLC, and ACE of the test sample was determined according to the following formula (A = peak area of hypuric acid extracted from the test sample, B = peak area of hippuric acid extracted from 10% dimethyl sulfoxide). The inhibition rate was determined.

      ACE inhibition rate (%) = (1−A / B) × 100

3) Test results The results are shown in Table 4.

  As shown in Table 4, the depolyphenol-treated NAP of the cocoa nib extract, which is an angiotensin I converting enzyme inhibitor of the present invention, showed a significantly higher ACE inhibition rate than the marine peptide.

  From the above studies, it has been clarified that the angiotensin I converting enzyme inhibitor of the present invention exhibits a significantly higher ACE inhibitory effect than a sadden peptide that is effective in lowering blood pressure.

  EXAMPLES Hereinafter, although an Example is given and this invention is demonstrated further in detail, it does not restrict | limit the range of the product of this invention by them.

  Using NAPS obtained in Test Example 4, tablets were prepared according to the following formulation.

D-mannitol 44.0 parts Lactose 40.0 parts Crystalline cellulose 10.0 parts Hydroxypropyl cellulose 5.0 parts
Fractionated NAPS 1.0 part

  Using the NBPS obtained in Test Example 4, a lozenge was prepared according to the following formulation.

Glucose 72.3 parts Lactose 19.0 parts Gum arabic 6, 0 parts Fragrance 1.0 parts Sodium monofluorophosphate 0.7 parts
Fraction NBPS 1.0 part

  Using NBPD obtained in Test Example 4, chewing gum was prepared according to the following formulation.

Gum base 20.0 parts Sugar 55.0 parts Glucose 15.0 parts Minamata 9.0 parts Fragrance 0.5 parts
Fraction NBPD 0.5 part

  Using JAPS obtained in Test Example 4, candy was prepared according to the following formulation.

Sugar 50.0 parts Minamata 34.0 parts Fragrance 0.5 Partial painting JAPS 0.5 parts
15.0 parts of water

  Using JBPD obtained in Test Example 4, tablet confectionery was prepared according to the following formulation.

Sugar 76.4 parts Glucose 19.0 parts Sucrose fatty acid ester 0.2 parts Fragrance 0.2 Partial fraction JBPD 0.1 part
4.1 parts of water

  Using JBPS obtained in Test Example 4, gummy jelly was prepared according to the following formulation.

Gelatin 60.0 parts Chicken pox 23.0 parts Sugar 8.5 parts Vegetable oil 4.5 parts Mannitol 2.9 parts Lemon juice 1.0 part
Fraction JBPS 0.1 part

  Using NBPD obtained in Test Example 4, chocolate was prepared according to the following formulation.

Powdered sugar 40.8 parts Cocoa bitter 20.0 parts Whole milk powder 20.0 parts Cocoa butter 17.0 parts Mannitol 1.0 Partial fraction NBPD 1.0 part
Fragrance 0.2 parts

  Using JAP obtained in Test Example 2, sherbet was prepared according to the following formulation.

Orange juice 25.0 parts Sugar 25.0 parts Egg white 10.0 Partial painting JAP 2.0 parts
38.0 parts of water

  Using the NBP obtained in Test Example 2, biscuits were prepared according to the following formulation.

Soft flour grade 1 25.0 parts Medium flour grade 1 22.0 parts Whitened sugar 5.0 parts Salt 1.0 parts Glucose 1.0 parts Palm shortening 12.0 parts Sodium bicarbonate 0.2 parts Sodium bisulfite 0. 2 parts Rice flour 2.0 parts Whole milk powder 1.0 parts Substitute milk powder 0.6 Partial fraction NBP 1.0 parts
29.0 parts of water

  Using JAP obtained in Test Example 2, an ice cream was prepared according to the following formulation.

Nonfat dry milk 50.0 parts Fresh cream 25.0 parts Sugar 10.0 parts Egg yolk 10.0 Partial painting JAP 1.0 part Fragrance 0.1 part
3.9 parts of water

  Using JBP obtained in Test Example 2, a powder was prepared according to the following formulation.

Corn starch 55.0 parts Carboxymethyl cellulose 40.0 parts
Fraction JBP 5.0 parts

  Using the NC obtained in Test Example 1, chewing gum was prepared according to the following formulation.

Gum base 20.0 parts Sugar 55.0 parts Glucose 15.0 parts Minamata 9.0 parts Fragrance 0.5 parts
Extract NC 0.5 parts

  Using JAP obtained in Test Example 1, ice cream was prepared according to the following formulation.

Nonfat dry milk 50.0 parts Fresh cream 25.0 parts Sugar 10.0 parts Egg yolk 10.0 parts Extract NA 1.0 part Fragrance 0.1 parts
3.9 parts of water

  Using NP obtained in Test Example 3, a beverage was prepared according to the following formulation.

Orange juice 30.0 parts Isomerized sugar 15.0 parts Citric acid 0.1 part Vitamin C 0.1 Partial fraction NP 0.1 part Fragrance 0.1 part
54.6 parts of water

  Using JP obtained in Test Example 3, a jam was prepared according to the following formulation.

Pulp 4.0 parts Sugar 65.0 parts Clarified fruit juice 25.0 parts Citric acid 0.5 Partial fraction JP 2.0 parts
3.5 parts of water

Claims (6)

The extract of cacao nibs and / or cacao nibs with water, methanol, ethanol, ethyl acetate, or a mixed solvent thereof is subjected to one or more depolyphenol treatments selected from the group consisting of polyvinylpyrrolidone, methacrylate polymers and polyphenol oxidase. An angiotensin I converting enzyme inhibitor. One or more depolyphenol treatments selected from the group consisting of polyvinyl pyrrolidone, methacrylate polymer, and polyphenol oxidase in cocoa jam and / or an extract of cocoa jam with water, methanol, ethanol, ethyl acetate, or a mixed solvent thereof. An angiotensin I converting enzyme inhibitor.   An angiotensin I converting enzyme inhibitor obtained by further subjecting the depolyphenol-treated product according to claim 1 or 2 to an amino acid composition concentration treatment by column fractionation. Treating cacao nib and / or an extract of cacao nib with water, methanol, ethanol, ethyl acetate, or a mixed solvent thereof with one or more depolyphenols selected from the group consisting of polyvinylpyrrolidone, methacrylate polymer and polyphenol oxidase A method for producing an angiotensin I converting enzyme inhibitor comprising: One or more depolyphenol treatments selected from the group consisting of polyvinyl pyrrolidone, methacrylate polymer, and polyphenol oxidase from cocoa jam and / or an extract of cocoa jam with water, methanol, ethanol, ethyl acetate, or a mixed solvent thereof. A method for producing an angiotensin I converting enzyme inhibitor comprising:   The method for producing an angiotensin I converting enzyme inhibitor according to claim 4 or 5, further comprising concentrating the amino acid composition by column fractionation on the depolyphenol-treated product.