JPH10114674A - Promoter of mineral absorption and preventive for hyperlipemia and hypercholesterolemia - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a promoter of mineral absorption capable of promoting the intraintestinal absorption of calcium and iron contents and preventing hyperlipemia and hypercholesterolemia, and useful for a food material, etc., by including a vegetative peptide having a prescribed molecular weight and containing a prescribed amount of acidic amino acid. SOLUTION: This promoter of mineral absorption comprises (A) a vegetative peptide having 200-6000 dalton molecular weight, containing >=20mol% acidic amino acid, and obtained by hydrating a vegetative protein selected from corn gluten, wheat gluten and corn casein by an enzyme such as papain and bromelain under a condition of 30-80 deg.C and pH 40-8.0 for 20-30hr, and further preferably (B) one or more kinds of a citric acid (salt), a sodium salt, a potassium salt and a calcium salt.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a novel use of a gluten peptide produced by hydrolyzing a vegetable protein. More specifically, the present invention relates to a mineral absorption promoter containing a gluten peptide as an active ingredient. Hyperlipidemia (h
The present invention relates to a prophylactic agent for hyperlipidemia and hypercholesterolemia.

[0002]

BACKGROUND OF THE INVENTION The nutrition of food proteins has been emphasized in terms of their basic functions: nitrogen sources, amino acid sources, and intermediates in the metabolism of many proteins that function in the body. Therefore,
In the nutritional evaluation of food proteins, qualitative and quantitative determination of amino acids constituting the protein has been centered.

[0003] Recently, research results on the function of peptides derived from food proteins in addition to the basic functions of food proteins as amino acid sources have been published, and new recognition has been gained about the nutritional value of food proteins. . Regarding the physiological functions of peptides derived from food proteins, firstly, there are many peptides having physiological functions in the living body, and it was revealed that they were generated from precursor proteins, and secondly, Protein 1
Since the structure similar to the physiologically functional peptide in the living body exists in the amino acid sequence of the food protein as the secondary structure is gradually revealed, many peptides derived from the food protein It has been confirmed that it has functions.

[0004] In the process of digesting proteins, it has been revealed that peptides having various molecular weights are produced in the digestive tract in a variety of manners according to the types of proteins, and these peptides perform various physiological functions. . For example, in the process of digesting casein, a milk protein,
Casein phosphopeptide (casein phosphph)
optide (CPP) was produced, and it was reported that this peptide acts as a calcium absorption enhancer. Some peptides produced during the digestion process are angiotensin converting enzymes.
Nvertingenzyme (ACE) activity has been reported to exhibit an inhibitory effect, and opioids (opioi) have been reported.
Peptides derived from various food proteins that exhibit the physiological function of de) have also been reported.

Further, zein (zei), a kind of wheat protein such as gluten or corn protein, is used.
Angiotensin-converting enzyme inhibitor (angiotensin) which has a function of suppressing mucosal degeneration of the small intestine using a protein hydrolase from n) and is used for the treatment of hypertension
converting enzyme inhibi
tor).

In particular, peptides produced by decomposing gluten, which is a corn protein, using a protein hydrolase are widely used as fatigue recovery and enteral nutrition foods. The corn gluten used for the production of the peptide is produced as a by-product during the production of starch by processing corn, and this gluten is added as a protein source during the production of soy sauce, or added as a protein source to animal feed. It has been used.

On the other hand, among the minerals essential for the human body,
Calcium and iron are known as nutrients that are particularly deficient in the body. As life becomes more abundant, caloric intake of proteins and fats exceeds daily requirements, causing various adult diseases, but calcium and iron intakes continue to be 90% of daily requirements. Induces osteoporosis and severe anemia. As described above, there is a method of ingesting more than the required daily amount to replenish calcium and iron deficiency in the body, but digestion disorders etc. occur and burden the stomach. It is efficient to increase the mineral absorption rate.

[0008] On the other hand, among the recently increasing adult diseases, diseases related to the circulatory system are the most common, and a method of preventing and treating the diseases by eating limited food, that is, dietary therapy is of interest to the public. It has become a target. For example, high concentrations of lipids and cholesterol in blood have been pointed out as major causes of circulatory diseases such as brain diseases, arteriosclerosis, and high blood pressure. However, diets by regulating food intake can control fat and cholesterol levels, which are affected by cholesterol levels, fat types and amounts, hydrocarbon types, vitamins, minerals, and the like. Consistent results have not been presented according to the experimental period, experimental conditions, experimental animal species, experimental animal age, etc., but due to the close relationship between protein type and lipid metabolism, casein increases serum cholesterol levels and soybeans. It has been revealed that the protein has an effect of reducing serum cholesterol level.

[0009] As a mechanism of the expression of the action, changes in fat absorption depending on the type of protein and amino acid composition, lysine / arginine ratio, sulfur-containing amino acid, and excretion of fat and cholesterol in feces are considered as fat and cholesterol. Have been suggested to contribute to the regulation of Recently, proteins and peptides produced during the digestion process are thought to interact with bile, lipids, cholesterol, and the like. For example, the effect of lowering lipid and cholesterol levels by soy protein is
It is believed to be the effect of high molecular weight peptides produced during the digestion process. And, due to the nutritional and physiological superiority of dietary peptides, development of novel peptides used in the form of food or medicine is desired.

[0010] In view of the above problems, the present inventors have conducted intensive studies and, as a result, have developed a novel peptide which can be produced by hydrolyzing an inexpensive and easily available vegetable protein with an enzyme.

In general, vegetable peptides have been produced by hydrolyzing proteins derived from soybean, wheat, corn and the like with enzymes. For example, Japanese Unexamined Patent Publication
In -204900, a peptide produced by a process of treating a vegetable protein with a starch hydrolyzing enzyme, heating the thus obtained substance under a high alkali, adding an alkaline protease, and then removing an aromatic amino acid. Contained substances are disclosed. In the production method, a step of removing an aromatic amino acid is essential in order to increase the relative content of branched amino acids known to have an effect of improving muscle exercise function and to remove bitterness. Have been. In addition, the above publication discloses that since the peptide-containing substance is rapidly absorbed into the digestive tract in the same manner as amino acids, it can be used for various uses for replenishing nutrition when physical strength decreases due to intense exercise and the like. It suggests.

[0012]

SUMMARY OF THE INVENTION The main object of the present invention is to provide:
An object of the present invention is to provide a calcium and iron mineral absorption promoter containing, as an active ingredient, a peptide produced by hydrolyzing a vegetable protein. It is a further object of the present invention to provide a preventive agent for hyperlipidemia and hypercholesterolemia, which contains, as an active ingredient, a peptide produced by hydrolyzing a vegetable protein.

[0013]

Means for Solving the Problems The present inventors have found that a peptide prepared by hydrolyzing a vegetable protein, which is a by-product in the production of starch, with an enzyme is 200 to 6,000.
Having a molecular weight of daltons and containing at least 20 mol% of acidic amino acids, and finding that the peptide promotes intestinal absorption of calcium and iron and is useful for hyperlipidemia and hypercholesterolemia, The present invention has been completed. That is, the present invention is a mineral absorption promoter comprising a vegetable peptide having a molecular weight of 200 to 6000 daltons and containing an acidic amino acid of 20 mol% or more. Also, the present invention
A preventive agent for hyperlipidemia and hypercholesterolemia comprising a vegetable peptide having a molecular weight of 200 to 6000 daltons and containing an acidic amino acid of 20 mol% or more.

[0014]

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described more specifically. Vegetable proteins, such as corn and wheat gluten, are added to a proteolytic enzyme, such as papain or bromelain, at a temperature of 30 to 80 ° C., preferably 50 to 60 ° C., and an acidic pH of 4.0 to 8.0. To a neutral condition, preferably pH 6.0, for 20 to 30 hours to hydrolyze, have a molecular weight of 200 to 6000 daltons and 20 mol%
Gluten peptides containing the above acidic amino acids were prepared. It was confirmed that the gluten peptide thus prepared (hereinafter referred to as “gluten hydrolyzate”) has an ability to inhibit the formation of calcium and iron precipitates in vitro. Furthermore, in rats that have taken the gluten hydrolyzate, the amount of precipitated calcium is small,
It was confirmed that the soluble calcium concentration in the small intestine was high.

Therefore, in the present invention, gluten, gluten hydrolyzate, gluten hydrolyzate or gluten is used as a nitrogen source in order to confirm whether the gluten hydrolyzate can promote intestinal absorption of calcium and iron in vivo. The diet to which the supernatant or precipitate of the hydrolyzate was added was supplied to each group of rats. As a result, in all experimental groups, there was no statistically significant difference in average body weight, daily food intake, daily weight gain, and feed efficiency at the time of starting and ending food supply, but gluten hydrolysis was not observed. In the experimental group that consumed the diet containing the hydrolyzate or gluten hydrolyzate supernatant, it was shown that the absorption rate of calcium and iron was increased. Also, it can be seen that the higher the concentration of the gluten hydrolyzate, the higher the calcium and iron absorption rates. This indicates that the gluten hydrolyzate promotes intestinal absorption of calcium and iron. Furthermore, it was confirmed that the calcium and iron precipitation inhibiting ability of the gluten hydrolyzate was improved by adding citric acid, citrate, sodium salt, potassium salt, calcium salt and the like.

On the other hand, in order to examine whether the gluten hydrolyzate has a preventive effect on hyperlipidemia and hypercholesterolemia induced by dietary fat intake, adult rats contain high fat and high cholesterol. An experimental diet prepared such that casein, casein hydrolyzate, gluten and gluten hydrolyzate were respectively contained in the basic diet to be fed was fed, and the onset levels of hyperlipidemia and hypercholesterolemia were measured. Also, adult rats are fed a diet containing high fat, high cholesterol and casein, a protein source, to produce hyperlipidemic rats induced by dietary fat, and fed the experimental diet to hyperlipidemia. And the therapeutic effect of hypercholesterolemia was measured. In this experiment, arterial blood, liver, heart and feces were collected from each rat and the total lipid and cholesterol content was determined from these samples. The experiments performed in the present invention and the results are summarized below.

1) The effect of gluten hydrolyzate on the prevention of hyperlipidemia and hypercholesterolemia induced by dietary fat was examined. As a result, there was no difference in dietary intake between the experimental groups. Rats fed the hydrolyzate had relatively low weight gain. Further, the effect of gluten hydrolyzate on the treatment of hyperlipidemia and hypercholesterolemia induced by dietary fat was examined. As a result, there was no difference in dietary intake and body weight gain in each experimental group. 2) The serum total lipid concentration was highest in the casein group and lowest in the gluten hydrolyzate group.
Serum total lipid concentration was significantly affected by nitrogen source and hydrolyzate. As a result, gluten and its hydrolyzate reduce the effect of reducing hyperlipidemia (hypolipidemic).
effect). 3) The effect of ingesting gluten and its hydrolyzate on serum total cholesterol concentration was similar to the effect on serum total lipid concentration. That is, the serum total cholesterol concentration was significantly affected by the nitrogen source and the hydrolyzate. As a result, gluten and its hydrolyzate reduce hypercholesterolemia (hypocholesteremia).
olemic effect). In addition, it was revealed that the intake of protein hydrolyzate increased serum HDL-cholesterol level and decreased serum LDL-cholesterol level. 4) Total lipid and total cholesterol content in liver was significantly reduced by gluten or protein hydrolyzate intake, but total cardiac lipid and total cholesterol content was not affected by gluten or protein hydrolyzate intake . 5) Total lipid excretion and total cholesterol excretion in feces were significantly increased by gluten intake, but were not affected by gluten hydrolyzate intake.

From the above results, it was revealed that gluten and its hydrolyzate have a hyperlipidemia-reducing effect and a hypercholesterolemia-reducing effect. That is, gluten and its hydrolyzate can reduce liver serum fat concentration and fat content. Therefore, gluten and its hydrolyzate can be used as an active ingredient of food for prevention and treatment of diet-induced hyperlipidemia and hypercholesterolemia.

According to the present invention, gluten peptides that promote intestinal absorption of calcium and iron and prevent hyperlipidemia and hypercholesterolemia can be prepared from corn and wheat, which can be purchased at low cost and easily. Therefore, a mineral absorption promoter containing a gluten peptide as an active ingredient and a preventive agent for hyperlipidemia and hypercholesterolemia can be provided in large quantities at low cost.

The mineral absorption enhancer of the present invention containing gluten hydrolyzate as an active ingredient and the preventive agent for hyperlipidemia and hypercholesterolemia are orally administered in the form of a pharmaceutically acceptable bile-containing preparation. It can be administered or injected. For oral administration, the peptide is a tablet,
It can be formulated into solid preparations such as pills, granules, powders, capsules and the like, and can also be formulated into liquid preparations such as solutions, suspensions and emulsions. Pharmaceutical preparations for oral administration comprise the active peptide or peptides, (a) filler and bulking agent (eg, starch, lactose, sucrose, glucose, mannitol and silica), (b) a binder (eg, carboxymethylcellulose, Alginates, gelatin and polyvinylpyrrolidone), (c) diluents (eg, glycerin), (d) disintegrants (eg, agar-agar, calcium carbonate and sodium carbonate),
(E) dissolution inhibitor (for example, paraffin), (f) absorption promoter (for example, quaternary ammonium compound), (g)
Wetting agents (eg, cetyl alcohol or glyceryl monostearate), (h) adsorbents (eg, kaolin, bentonite), (i) lubricants (eg, talc,
Calcium stearate, magnesium stearate and solid polyethylene glycol), (j) coloring agents,
(K) a seasoning, (l) a sweetener, or a mixture of the substances listed in (a) to (l) together with common excipients. When the preparation is used for parenteral administration, the preparation is formulated for injection (eg, intravenously). When prepared for injection, sterile solutions and emulsions can be made isotonic with blood. Suspensions may contain, in addition to the active peptide or peptides, preservatives, stabilizers, solubilizers, wetting agents, salts that alter osmotic pressure, or buffers. Less than,
The present invention will be described more specifically with reference to examples, but the scope of the present invention is not limited to these examples.

【Example】

Example 1 Preparation of Gluten Hydrolyzate To prepare a gluten hydrolyzate, 1.5 kg of corn gluten or wheat gluten was added to 1 L of water.
Papain or bromelain (10 to 150 g) was added and reacted under the conditions of a temperature of 50 ° C. and a pH of 6.0.
During the reaction, pH 6.0 was kept constant by adding sodium hydroxide or hydrochloric acid by a pH controller installed in the reactor. The gluten hydrolyzate produced by reacting for 24 hours contains not less than 20 mol% of acidic amino acids and
It has a molecular weight of 0 to 6000 daltons. This gluten hydrolyzate was centrifuged, separated into a supernatant and a precipitate, and each was collected, and then used in the next example.

Example 2 : 10% gluten hydrolyzate
Feeding Rats Containing (w / w) -Containing Diet Twenty-four 24-week-old male Sprague-Dawley rats were divided into four groups, and 10% (w / w) of soybean protein was added as a nitrogen source to gluten, Gluten hydrolyzate, supernatant of gluten hydrolyzate or precipitate of gluten hydrolyzate
Different diets containing 0% (w / w) were allowed the rats of each group to eat freely for 4 weeks. At this time, the rats were separated and reared in a shoe-box cage, and the temperature of the rearing room was 22 ± 2 ° C. and the relative humidity was 6 ° C.
Light conditions from 6 am to 6 pm were kept constant at 5 ± 5%. In addition, semi-purified diet (se
mi-purified diet) was used. Its components and composition are AIN-76 (American In
state of Nutrition Stan
dards for Nutritional Stu
dies Report, J. et al. Nutr. , 107: 1
340-1348 (1977)).

[0023]

[Table 1]

Example 2-1 : Measurement of weight gain of rats As a result of investigating the daily food intake and weight gain of rats, there was no significant difference in the food intake and weight gain of each experimental group. However, in the four groups, a slight decrease in body weight gain was observed due to a decrease in daily food intake (see FIG. 1).

Example 2-2 : Absorption of calcium and iron in rats The absorption of calcium and iron in rats was measured.
And Table 3. At this time, the concentration of calcium was measured by a general method, and the concentration of iron was measured using a reagent for quantitative analysis of iron (Wako, Japan). In this experiment, the stock (mg / dl or μg / ml) was determined by subtracting the amount of daily food intake minus the amount excreted in feces and urine. Apparent absorption rate (mg / dl or μg / m
l) was determined by subtracting the daily food intake from the fecal output. Storage and apparent absorption were calculated as percentages of daily food intake, respectively.

[0026]

[Table 2]

As shown in Table 2, the apparent absorption rate (%) of calcium is about 38% in the group fed the gluten-containing diet, but the gluten hydrolyzate is contained in one group. Approximately 5 in the case of two groups fed
4%, which corresponds to an increase in apparent absorption of about 42%. For the three groups fed a diet containing gluten hydrolysate supernatant, this is about 50%, which is about 31%.
Corresponds to an increase in the apparent absorption rate. Therefore, it was revealed that the gluten hydrolyzate promotes intestinal absorption of calcium.

[0028]

[Table 3]

The apparent absorption rate (%) of iron is about 9% in the case of the group fed the gluten-containing diet, but is about 3% in the group fed the gluten hydrolyzate supernatant. About 16% for the group, which is about 8%
This corresponds to an increase in apparent absorption of 0%. Therefore, it was revealed that gluten hydrolyzate promotes intestinal absorption of iron in the same manner as calcium.

Example 3 Gluten hydrolyzate was 2% (w
/ W) Feeding of the containing diet to rats Fourteen-week-old male Sprague-Dawley rats were divided into three groups, and 18% soy protein was used as a nitrogen source.
In addition to (w / w), different diets containing 2% (w / w) gluten, gluten hydrolyzate or gluten hydrolyzate supernatant were allowed to freely ingest the rats of each group for 4 weeks. Rats were bred in a manner similar to Example 2 and the components and composition of the experimental diet are shown in Table 4 below.

[0031]

[Table 4]

Example 3-1 : Measurement of Weight Gain of Rats The average weight, daily food intake, daily weight gain, and diet efficiency of rats during feeding were investigated. As a result, although no statistically significant difference was shown in each experimental group, the group fed the gluten hydrolyzate supernatant showed a slightly higher average body weight at the end of feeding (see FIG. 2). ).

Example 3-2 : Measurement of absorption rate of calcium and iron in rat The absorption rate of calcium and iron in rat was measured.
And Table 6. The concentrations of calcium and iron were measured by the same method as in Example 2-2. In this experiment, the storage amount (mg / dl or μg / ml), the apparent absorption rate (mg / dl or μg / ml) and their percentage were calculated as in Example 2-2.

[0034]

[Table 5]

[0035]

[Table 6]

The apparent absorption (%) of calcium and iron in Tables 5 and 6 was compared with the apparent absorption (%) of calcium and iron in Tables 2 and 3.
% (W / w) gluten hydrolyzate supernatant, the group fed 10% (w / w) gluten hydrolyzate was lower than the group fed 2% (w / w) gluten. A 10 to 20% increase in absorption compared to was observed.

Example 4 : Improvement of calcium precipitation inhibiting ability by citric acid or sodium chloride To improve calcium precipitation inhibiting ability by gluten hydrolyzate, 0.1% (w / v) gluten hydrolyzate was added to 0.1% (w / v) gluten hydrolyzate. After adding 1 M citric acid, malic acid or sodium chloride and reacting at 30 to 80 ° C. for 1 to 3 hours, the mixture was spray-dried and powdered. To the powder thus prepared, 0.2 ml of distilled water was added to dissolve the powder, and 1 m of a 0.01 M CaCl ₂ .2H ₂ O solution was added thereto.
and 2 ml of a 0.02 M phosphate buffer solution (pH 7.0), incubated at 37 ° C. for 1 hour, and then at 12,000 rpm for 5 minutes in a small centrifuge equipped with a 0.1 μm ultrafiltration membrane. The supernatant was obtained by centrifugation. Then, the calcium concentration in the supernatant was measured (see Table 7).

[0038]

[Table 7] As shown in Table 7 above, it was revealed that the addition of citric acid or sodium chloride to the gluten hydrolyzate promoted the ability to inhibit calcium precipitation.

Example 5 : Preventive effect of gluten hydrolyzate on hyperlipidemia and hypercholesterolemia Male Sprague-Daw having an average body weight of about 160 g
32 ley rats were arbitrarily divided into 4 groups of 8 rats, and high fat (18% in the diet) and high cholesterol (1% in the diet)
%) Were fed an experimental diet containing casein (C), casein hydrolyzate (CH), gluten (G) and gluten hydrolyzate (GH) for 6 weeks, respectively, to obtain hyperlipidemia and hyperlipidemia. The onset level of cholesterol was measured. At this time, the rats were bred in the same manner as in Example 2, and a metabolic experiment was performed for 4 days before the rats were sacrificed. The experimental diet was a semi-purified diet, and its components and compositions are shown in Table 8 below. The experimental diet was purified tallow (Seo)
ul Agricultural Products
Co. , Korea) at a concentration of 18% (this concentration is NAS-
NRC (National Research Cou)
ncil, Nutrient Requirements
nts of Laboratory Animals
No. 10, NAS, Washington D.C.
C. (1972)), which is higher than the standard fat concentration of 5%. AIN) except for the addition in
Prepared according to -76. In addition, casein (Maeil
MilkCo. , Ltd. , Korea), gluten and gluten hydrolyzate were added to each diet at a concentration of 10%.
The nitrogen content of each experimental diet was the same, and PEG (polyethylene glycol) # 40 was used as a food movement marker in the gastrointestinal tract.
00 was included in the diet at 1%.

[0040]

[Table 8]

Feeding of the experimental animals was stopped for 12 hours before sampling and thereafter for 1.5 hours. One hour after the end of feeding,
Rats were anesthetized with ethyl ether and blood was collected from the carotid artery. After the blood was collected, the liver and the heart were excised to remove fat adhering to the organs, and then added to cold saline (0.9%).
% NaCl solution) to remove blood, weigh and collect feces 4 days before sacrifice of rats.
The collected blood is stored in a refrigerator (4 ° C) for 24 hours,
Serum was obtained by centrifugation at 3000 rpm for 20 minutes. Freeze-dryer (Freeze-Dryer 1)
8) After freeze-drying and crushing using (Labconco, USA), the dry weight was measured. At this time, all samples were stored frozen at -40 ° C or lower until used for analysis.

Example 5-1 : Body Weight and Food Intake The body weight, weight gain, food intake, and feed efficiency of the rats that consumed each experimental diet were measured and are shown in Table 9 below.
In this experiment, feed efficiency is the value obtained by dividing the weight gain by the food intake. As can be seen from Table 9, end weight,
There was a significant difference between each experimental group in weight gain and feed efficiency, but no difference in food intake. The casein and casein hydrolyzate intake groups showed higher weight gain than the gluten and gluten hydrolyzate intake groups, which was presumed to be due to the difference in the amino acid composition of the two proteins. The experimental data was analyzed using the SAS program, and the mean and standard error (mean ± SE) were determined. The significance of each treatment and the effect of the type of protein (nitrogen source) [(C + CH) :( G + GH)] and the effect of the hydrolyzate [(C + G) :( CH + GH)] are Duncan's multiple range test (Dumcan's multi
(p <0.05).

[0043]

[Table 9] 1) Mean ± SE 2) In all the tables below, different superscripts in the same column represent significantly different numerical values (p <0.05). 3) NS (Not Significantly Different): Not significantly different.

Example 5-2 : Concentrations of Total Lipids and Triglycerides in Serum The concentrations of total lipids and triglycerides in the serum of rats fed an experimental diet were measured and are shown in Table 10 below. In this experiment, the total lipid concentration in the serum was determined by Fringe.
And Dunn's method (Fringe, C. S. a.)
nd Dunn, R .; M. , Am. J. Clin. Pa
th. , 53: 89-92 (1980)) and the sulfo-phosphovanillin reaction (sulfo-phos).
The concentration of triglyceride in serum was measured by a colorimetric method using phovanillin reaction, and the concentration of triglyceride in serum was analyzed by the method of Biggs et al.
G. FIG. et al. , Clin. Chem. , 21:43
7-441 (1975)).

[0045]

[Table 10] * The values in parentheses indicate the percentage of the casein diet group (1 group).

As can be seen from Table 10 above, the serum total lipid concentration was significantly different between the experimental groups, but the serum triglyceride concentration was not significantly different. The serum total lipid concentration was significantly higher in the casein intake group than in the other experimental groups, and there was no significant difference in serum total lipid concentration among the other three groups. In particular, the gluten hydrolyzate intake group showed the lowest value. That is, 40% or more lower than the casein intake group and 22% compared to the gluten intake group.
It was low. On the other hand, a significant nitrogen source effect was observed in the serum lipid concentration. This is because the effect of the gluten and its hydrolyzate intake group was as low as almost 74% as compared with the effect of casein and its hydrolyzate. These results were consistent with many reports that serum lipid levels were significantly lower in the soy protein intake group than in the casein intake group. Further, when the effect of ingesting the hydrolyzate on the protein was examined, the hydrolyzate group was almost 75% less than the protein group.
%, Indicating a significant reduction effect. Therefore, the gluten hydrolyzate has a preventive effect against hyperlipidemia induced by a high-fat food containing high cholesterol due to the effect of reducing the concentration of gluten protein and its hydrolyzate with respect to the serum total lipid concentration. The conclusion was reached.

Example 5-3 : Concentration of Total Cholesterol and Fatty Protein Cholesterol in Serum Total cholesterol and high-density fat protein cholesterol (HDL) in serum were measured, and
The low density fat protein cholesterol (LDL) concentration and the atherosclerosis index (Ather
o-Index: AI) was calculated and shown in Table 11 below. In this experiment, the serum total cholesterol concentration was determined by the method of Zlatkis and Zak (Zlatkis, A. and Zak, B., Ana).
l. Biochem. , 29: 143-146 (196)
According to 9)), serum HDL-cholesterol was determined by an enzyme method kit (Young-Dong Pharmaceutical Co., Ltd., Korea)
It measured using.

[0048]

[Table 11] 1) LDL-cholesterol: total cholesterol- (HDL-cholesterol) -triglyceride / 5 2) Atherosclerosis index: (total cholesterol- (HDL-cholesterol)) / (HDL-cholesterol)

As can be seen from Table 11, the total cholesterol concentration in the serum is significantly different between the protein intake group and the hydrolyzate intake group, but not according to the type of protein. In particular, the casein hydrolyzate group showed a significant decrease of about 15% in serum total cholesterol concentration compared to the casein group, and also showed a decreasing tendency in the gluten hydrolyzate group as compared to the gluten group. However, no statistical significance was observed.
There was no significant difference between the casein intake group and the gluten intake group. On the other hand, the effect of the experimental diet on the serum HDL cholesterol concentration showed a significantly higher value in the hydrolyzate intake group than in the protein intake group (p <0.05). The group showed an increase of more than 40% compared to the casein or gluten intake groups. Therefore, in view of the fact that HDL cholesterol is known as a fat protein having an inhibitory effect on harmful factors of circulatory diseases, when a protein hydrolyzate, especially gluten hydrolyzate is ingested, circulating such as arteriosclerosis, hypertension, etc. It is thought to reduce the occurrence of organ diseases. Furthermore, LDL calculated from the concentration of total cholesterol, HDL cholesterol and triglyceride
The cholesterol concentration was significantly lower in the hydrolyzate intake group than in the protein intake group (p <0.05). The gluten intake group tended to be lower than the casein intake group, but there was no statistically significant difference. These results demonstrated that ingestion of protein hydrolyzate had the effect of reducing serum total cholesterol and increasing HDL cholesterol, which significantly reduced the atherosclerosis index.

Example 5-4 Content of Total Lipid, Total Cholesterol and Triglyceride in Liver Tissue The wet weight, dry weight and total lipid, total cholesterol and triglyceride content per liver tissue were measured.
The results are shown in Table 12 below. The method of Folch et al. (Folch et al., J. Biol. Chem., 226, 497-502 (195
After extracting the total fat from the tissue using (7)), the total cholesterol and triglyceride contents were determined in Example 5-2.
Quantification was performed in the same manner as in

[0051]

[Table 12]

As can be seen from Table 12, the live tissue weight and dry weight of the liver, the total lipid content, the total cholesterol content, and the triglyceride content all showed significant differences between the experimental groups. The liver weight showed a significant difference according to the type of protein, and the casein intake group was higher than the gluten intake group, but there was no difference between the protein intake group and its hydrolyzate intake group. Total lipid content,
The total cholesterol content and triglyceride content were significantly lower in the gluten intake group than in the casein intake group. The total lipid content and triglyceride content were lower in the hydrolyzate intake group than in the protein intake group, but were not significantly different. Therefore, it can be said that the type of protein and its hydrolyzate intake affect lipid metabolism in liver tissue. These results demonstrated that lipid metabolism in liver tissue was affected by the type of protein and nitrogen form, that is, the intake of protein hydrolyzate or gluten reduced lipid content in liver.

Example 5-5 Content of Total Lipid, Total Cholesterol and Triglyceride in Heart Tissue The weight, dry weight, and total lipid, total cholesterol and triglyceride content per dry weight of heart tissue were determined in Examples. Quantification was carried out in the same manner as in 5-4.
It was shown to.

[0054]

[Table 13]

As can be seen from Table 13, there was no significant difference between the experimental groups in the weight of the heart and the total cholesterol content, but there were significant differences in the total lipid and triglyceride contents between the experimental groups. That is, total lipids,
The triglyceride content did not show a significant difference according to the type of protein, but the hydrolyzate intake group showed a significantly lower value than the protein intake group (p
<0.05). In particular, when the gluten hydrolyzate was taken, the total lipid content of the heart tissue showed a significantly lower value.
On the other hand, the triglyceride content was significantly lower in each hydrolyzate intake group than in the protein intake group.

Example 5-6 : Excretion of lipids in feces The daily excretion of feces in each experimental group and the amounts of total lipids, total cholesterol and triglycerides contained therein were measured and are shown in Table 14 below. Was. The contents of cholesterol and triglyceride in feces were determined in the same manner as in Example 5-4.

[0057]

[Table 14]

As can be seen from Table 14, there was no significant difference in fecal excretion between the experimental groups. Only the total lipid showed a significant difference between the experimental groups, and the total lipid excretion was lower in the casein intake group than in the other groups. Gluten intake significantly increased total cholesterol and total lipid excretion. The excretion of total lipids, total cholesterol and triglycerides tended to increase slightly in the hydrolyzate-ingested group compared to the protein-ingested group, but there was no significant difference. Thus, it was found that ingestion of gluten all showed an increasing effect on the excretion of total lipids, total cholesterol and triglycerides.

Example 6 : Therapeutic effect of gluten hydrolyzate on hyperlipidemia and hypercholesterolemia Male rats having an average body weight of about 160 g (Sprag)
Ue-Dawleyrat) were fed a hyperlipidemia-inducing diet containing high fat, high cholesterol and casein as a protein source for 4 weeks to produce hyperlipidemia experimental model rats. Then, as in the fifth embodiment, 4
Four experimental diets were arbitrarily ingested for 4 weeks in groups, and the therapeutic effects of hyperlipidemia and hypercholesterolemia were measured and examined. Constituents and composition ratio of experimental diet, breeding conditions,
Sampling, sample analysis, and statistical analysis were performed in the same manner as in Example 5.

Example 6-1 : Body Weight and Dietary Intake The weight change, daily weight gain, dietary intake and feed efficiency associated with the intake of each experimental diet were measured and are shown in Table 15 below. As can be seen from Table 15, there was no difference between the experimental groups in body weight and food intake, with no nitrogen source effect and no hydrolyzate effect.

[0061]

[Table 15]

Example 6-2 : Concentrations of Total Lipids and Neutral Fats in Serum * The concentrations of total lipids and neutral fats in the serum of rats fed the experimental diet were measured and are shown in Table 16 below. .

[0063]

[Table 16] * The values in parentheses indicate the percentage of the casein diet group (1 group).

As can be seen from Table 16, there was no significant difference between the experimental diet groups in the serum triglyceride concentration. That is, serum triglyceride concentration was not affected by the type of protein and the form of nitrogen. However, the casein intake group showed significantly the highest serum total lipid concentration, and the gluten hydrolyzate intake group showed the lowest serum total lipid concentration, and the results were similar to the results of Example 5-2. . However, in this example, a high fat diet containing casein
After inducing hyperlipidemia by ingestion for 4 weeks, four experimental diets were consumed for 4 weeks in a therapeutic aspect.
A different evaluation must be made from the results of Example 5-2 where the weekly experimental diet was ingested. Therefore, it was demonstrated that the gluten hydrolyzate has a therapeutic effect on hyperlipidemia induced by a high-fat food containing high cholesterol by the effect of reducing the total serum lipid concentration of such gluten and its hydrolyzate. .

Example 6-3 : Serum Total Cholesterol and Fat Protein Cholesterol Concentrations Serum total cholesterol and high-density lipid protein cholesterol (HDL) concentrations were measured, and serum low density lipid protein cholesterol (LDL) was measured. ) Were calculated according to the Friedwald equation and the values are shown in Table 17.

[0066]

[Table 17] 1) LDL-cholesterol: total cholesterol- (HDL-cholesterol) -triglyceride / 5 2) Atherosclerosis index: (total cholesterol- (HDL-cholesterol)) / (HDL-cholesterol)

As can be seen from Table 17, the serum total cholesterol concentration showed a significantly higher value in the casein group and the lowest value in the gluten hydrolyzate group. Examining the effects of protein types and hydrolysates, the gluten or protein hydrolysate groups were compared to the casein groups.
Serum total cholesterol levels were significantly reduced from 55% to 69%. Such results suggest that gluten hydrolyzate can be used to treat hypercholesterolemia induced by a high fat diet. Looking at the effect of the experimental diet on the serum HDL cholesterol concentration, the gluten hydrolyzate intake group showed an increase of about 40% as compared to the casein intake group or the gluten intake group. Therefore, HD
Since L-cholesterol is known as a fat protein which has an inhibitory effect on risk factors for circulatory diseases, ingestion of protein hydrolysates, especially gluten hydrolysates, may cause circulatory diseases such as arteriosclerosis and hypertension. It is believed that the factor is reduced.

Further, L calculated from the total cholesterol, HDL cholesterol and triglyceride concentrations
The concentration of DL cholesterol tended to be lower in the hydrolyzate intake group than in the protein intake group, but did not show a statistically significant difference. Also,
Regarding the difference depending on the type of protein, the gluten intake group tended to show a lower value than the casein intake group, but there was no statistically significant difference. From these results,
Ingestion of protein hydrolyzate reduced serum total cholesterol levels and increased serum HDL cholesterol levels, demonstrating that these effects significantly reduced the atherosclerosis index.

Example 6-4 Content of Total Lipid, Total Cholesterol and Triglyceride in Liver Tissue The wet weight, dry weight and total lipid, total cholesterol and triglyceride content per liver tissue were measured.
The results are shown in Table 18 below.

[0070]

[Table 18]

As can be seen from Table 18, the weight of liver tissue did not differ between the experimental groups, but the total lipid, total cholesterol and triglyceride in the liver tissue showed significant differences between the experimental groups. . Although no difference was shown depending on the type of protein, the hydrolyzate intake group showed a significantly lower value than the protein intake group. Accordingly, it was demonstrated that the protein hydrolyzate had an effect of reducing total lipids, total cholesterol, and triglycerides in liver tissue when ingested for 4 weeks or more.

Example 6-5 : Total Lipid, Total Cholesterol and Triglyceride Content in Heart Tissue Wet weight, dry weight, total lipid per dry weight of heart tissue,
Total cholesterol and triglyceride content were measured and are shown in Table 19 below.

[0073]

[Table 19]

As can be seen from Table 19, only the triglyceride content showed a significant difference between the experimental groups, and the triglyceride content was significantly lower in the gluten intake group than in the casein intake group. In addition, no significant differences in the lipid content of heart tissue between the protein source and the form of nitrogen were shown between the experimental groups. Therefore, the total lipid content of the heart is hardly affected by the type of protein,
It was suggested that the effect of the hydrolyzate was hardly expected during the 4-week ingestion period.

Example 6-6 : Excretion of Total Lipid in Feces The daily excretion of feces and the contents of total lipids, total cholesterol and triglycerides in the experimental group were measured and are shown in Table 20 below. .

[0076]

[Table 20]

As can be seen from Table 20, total fat in feces,
The difference in the amount of triglyceride excreted between the experimental groups was significant. That is, each excretion amount was lowest in the casein intake group, but highest in the gluten hydrolyzate intake group. In addition, the total lipid and triglyceride excretion amount was significantly different depending on the type of protein, and each excretion amount was higher in the gluten intake group and the hydrolyzate intake group than in the casein intake group and the hydrolyzate intake group. . For total cholesterol excretion,
The gluten intake group tended to have higher excretion in feces than the casein intake group, but there was no significant difference. Therefore, it was demonstrated that the ingestion of gluten has the effect of increasing the excretion of total fat, total cholesterol and triglyceride in feces.

Further, the gluten hydrolyzate was subjected to a safety test. The gluten hydrolyzate of the present invention was administered to 4-week-old male and female rats (Sprague-Dawley).
(rat) was administered orally in an amount of 10 g per kg of body weight (maximum dose), and no harmful effects were detected.

[0079]

As described and demonstrated in detail above, the present invention has been produced by enzymatic hydrolysis of a vegetable protein.
Provided are a mineral absorption promoter and a preventive agent for hyperlipidemia and hypercholesterolemia containing, as an active ingredient, a peptide having an acidic amino acid in an amount of 20 mol% or more based on all amino acids and a molecular weight of 200 to 6,000 daltons.
According to the present invention, a gluten peptide is useful as a functional food material for promoting intestinal absorption of calcium and iron and preventing hyperlipidemia and hypercholesterolemia induced by ingestion of foods containing high fat and high cholesterol. It was recognized that.

[Brief description of the drawings]

FIG. 1 shows that gluten, gluten hydrolyzate, gluten hydrolyzate supernatant or gluten hydrolyzate precipitate is 10%
It is a graph which shows the weight with respect to breeding time of the rat which ingested the diet containing the ratio of (w / w).

FIG. 2 shows that gluten, gluten hydrolyzate, gluten hydrolyzate supernatant or gluten hydrolyzate precipitate is 2%
It is a graph which shows the weight with respect to breeding time of the rat which ingested the diet containing the ratio of (w / w).

──────────────────────────────────────────────────の Continued on the front page (51) Int.Cl. ⁶ Identification code FI A23L 1/305 A61K 31/19 A61K 31/19 33/00 33/00 33/06 33/06 35/78 AGAU // A61K 35 / 78 AGA C07K 14/415 C07K 14/415 A61K 37/02 ADD (72) Inventor Pei Tengho 432-4, Tamin-dong, Yuseong-gu, Daejeon, Republic of Korea Taichung No. 432-4, Tamin-dong, Yuseong-gu, Daejeon, Republic of Korea No. 108, Building No. 706 (72) Inventor Li Len-shu, No. 904, Myo-gu, 941, Myoka, Yamamoto-dong, Gunpo-si, Gunpo, Gyeonggi-do, Republic of Korea No. 904 (72) Inventor Park Ehime Princess 121, 2102, Bukseong-dong, Sejong-dong, Gunpo-si, Gyeonggi-do, Republic of Korea (72) Inventor Wu Zuhan 5th apartment, No. 607, 5-dong Apartment, Daelim 3-dong, Yeongdeungpo-gu, Seoul, Republic of Korea

Claims (10)

[Claims] 1. A mineral absorption enhancer comprising a vegetable peptide having a molecular weight of 200 to 6000 daltons and containing at least 20 mol% of acidic amino acids. 2. The mineral absorption enhancer according to claim 1, wherein the mineral is calcium or iron. 3. The method according to claim 1, wherein the vegetable peptide is prepared by enzymatically hydrolyzing a vegetable protein selected from the group consisting of corn gluten, wheat gluten and corn zein. Mineral absorption enhancer. 4. The hydrolysis by the enzyme is carried out at a temperature of 30 to 80 ° C. and a pH of 4.0 to 8.0.
The mineral absorption enhancer according to claim 3, wherein the treatment is performed for 0 to 30 hours. 5. The mineral absorption enhancer according to claim 4, wherein the enzyme is papain or bromelain. 6. A citric acid, a citrate, a sodium salt,
The mineral absorption enhancer according to claim 1, further comprising one or more substances selected from the group consisting of potassium salts and calcium salts. 7. A preventive agent for hyperlipidemia and hypercholesterolemia, comprising a plant peptide having a molecular weight of 200 to 6,000 daltons and containing an acidic amino acid of 20 mol% or more. 8. The method according to claim 7, wherein the vegetable peptide is prepared by enzymatically hydrolyzing a vegetable protein selected from the group consisting of corn gluten, wheat gluten and corn zein. For the prevention of hyperlipidemia and hypercholesterolemia. 9. The enzyme hydrolysis is carried out at a temperature of 30 to 80 ° C. and a pH of 4.0 to 8.0.
The preventive agent for hyperlipidemia and hypercholesterolemia according to claim 8, which is carried out for 0 to 30 hours. 10. The preventive agent for hyperlipidemia and hypercholesterolemia according to claim 9, wherein the enzyme is papain or bromelain.