JP2945994B2 - Nourishing composition - Google Patents

Description

Description: BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a nutritional supplement composition, and more particularly to a nutritional supplement or therapeutic food for liver disease, muscular dystrophy patients, and for invasion of living organisms. Nutritional supplement compositions.

2. Description of the Related Art In the case of patients with chronic liver failure such as cirrhosis, toxic substances generated in the body due to the formation of short-circuits inside and outside the liver and liver dysfunction bypass the liver parenchyma, making it difficult to detoxify. In addition to abnormalities, it is said that it eventually causes metabolic abnormalities in the brain and easily falls into a coma.

In 1974, Fischer measured plasma amino acid levels in patients with severe liver disease, and found a marked increase in aromatic amino acids (AAA) and methionine, including phenylalanine, tyrosine, free tryptophan, and valine, especially in patients with cirrhotic hepatic coma. It has been reported that abnormalities of amino acid metabolism are the main cause of hepatic encephalopathy, because the decrease in branched-chain amino acids (BCAA) such as leucine and isoleucine was observed. An amino acid infusion in which aromatic amino acids are restricted and the content of branched-chain amino acids is increased for the purpose of improving hepatic encephalopathy by correcting the amino acid imbalance in plasma caused by abnormal metabolism of amino acids (Fisher liquid) was developed. Since this amino acid infusion is excellent in awakening effect for hepatic encephalopathy patients, its usefulness is highly evaluated and widely used in clinical applications.

Patients with liver cirrhosis after awakening of hepatic encephalopathy are required to prevent recurrence of hepatic encephalopathy and improve nutritional status.However, even at the time of hypoalbuminemia, protein-limited diet with emphasis on the former is inevitable. However, such conventional therapies cannot correct the amino acid imbalance in plasma which leads to hepatic encephalopathy, cannot improve the nutritional status, and cannot improve the physical activity of the patients themselves. In addition, prolonged restriction of protein intake further exacerbates hypoalbuminemia.

For patients with cirrhosis, it is originally considered that a high calorie, high protein, and high vitamin diet is desirable for aggressive recovery. Therefore, not only treatment or prevention of hepatic encephalopathy but also nutritional considerations in patients with cirrhosis having or likely to develop hepatic encephalopathy are extremely important in patient management. Inadequate nutritional management is always at risk of developing hepatic encephalopathy. In view of these implications, administration by infusion is naturally limited, and it has been eager to develop a formulation for maintenance therapy after awakening of hepatic encephalopathy, which contributes to both improvement of hepatic encephalopathy symptoms and improvement of nutritional status.

From such a viewpoint, for the purpose of correcting amino acid imbalance in plasma and improving nutritional status, it is necessary to limit aromatic amino acids according to Fischer's theory, And nutritional supplements containing various nutrients have been studied. However, such nutritional supplements have the disadvantage that the osmotic pressure is too high due to the use of amino acids as a protein source,
Diarrhea occurs when ingested. Furthermore, there are unpleasant odors and bitterness peculiar to amino acids, and this point is also a problem.

On the other hand, muscular dystrophy (PMD) is a group of diseases having a primary disease in the muscle itself, mainly characterized by progressive degeneration of skeletal muscle, and is a disease that develops on a genetic basis. Examination of the amino acid composition in the blood of patients with muscular dystrophy shows that the ratio of essential amino acids to non-essential amino acids (E / N ratio), which is an indicator of undernutrition, is low even when the protein intake sufficiency rate is not necessarily poor. Abnormality of protein amino acid metabolism is considered. In addition, it has been reported that the concentration of branched-chain amino acids in blood is lower than that in normal humans (Kumiko Hirano, Bulletin of the Faculty of Life Sciences, Osaka City University, 34; 253-259, 1986). Suggests the need to increase the concentration of branched-chain amino acids in soil. Furthermore, among muscular dystrophies, it has been reported that in patients with amyotrophic lateral sclerosis (ALS), the effect of suppressing the decrease in muscular strength was confirmed by branched-chain amino acid supplementation (Plailakis A, et al: Lancet, May7; 1015-1018, 198
8).

In order to prolong the life of muscular dystrophy patients, it is necessary to provide sufficient energy and protein supplements and to delay weight loss.However, it is not easy to increase the diet for patients with occlusal disorders, etc. High energy, high protein replenishment at times is required.

In addition, a typical metabolic change at the time of stress load, particularly at the time of invasion of a living body such as surgical invasion, wound, burn, and sepsis is catabolism of protein metabolism. In other words, body proteins such as skeletal muscle, visceral smooth muscle, and subcutaneous collagenous tissue are decomposed,
At the same time as being consumed as an energy source, it is reused for the biosynthesis of acute-phase proteins (fibrinogen, haptoglobin, lysosomal enzymes, etc.) required for recovery, but is generally inclined toward proteolysis. As a result, the amount of urinary nitrogen excretion increases, and the nitrogen balance exhibits a negative value, causing weight loss and weakening the resilience.

ALGoldberg and colleagues focused on branched-chain amino acids (BCAAs), which are commonly used in muscle, and in a series of in vitro experiments using rat diaphragm muscle, BCAA exerted an inhibitory effect on muscle protein degradation or synthesis. It was reported for the first time that it had a promoting effect.

Blackburn (GLBlackburn) et al. And Fisher (JEFischer) show that since the late 1970s, using a variety of invasive model animals and administering amino acid infusions with a high BCAA ratio can improve protein metabolism under stress. Was.

Based on the results of these tests, BCAA
A high-concentration amino acid transfusion was trial-produced and examined clinically, and a remarkable nutritional effect was also confirmed clinically under stress.

From these findings, it is desired to develop a nutritional food with a high content of branched-chain amino acids for nutritional supplements for patients with muscular dystrophy or in the case of invasion of living organisms, but as described above, amino acids were added as a conventional protein source. Nutritional supplements are problematic in that they have too high osmotic pressure, unusual odor and bitterness peculiar to amino acids, and are not suitable as oral nutritional supplements for humans.

The present invention has been made in view of the above-mentioned problems of the prior art, and has as its object the capability of enteral administration such as oral and tube administration, digestion and absorption being performed physiologically, and high osmotic pressure. A nutritional supplement composition that can avoid the occurrence of diarrhea due to that and has no irritation or off-flavor.
An object of the present invention is to provide a nutritional supplement composition that can correct amino acid imbalance in blood and can improve nutritional status.

Means for Solving the Problems The present inventors have conducted intensive studies in order to achieve the above object. As a result, from a raw material containing corn protein, by a specific treatment, a peptide having a molecular weight of 200 to 4000 as a main component, in the amino acid composition, abundantly containing branched-chain amino acids, and containing almost no aromatic amino acids It has been found that a compound-containing substance can be obtained, and based on this, a predetermined amount of the peptide-containing substance is blended, and a composition in which the total amount of nitrogen compounds as a protein source, the amount of carbohydrate and lipid are adjusted to a predetermined ratio is ,
It has been found that it is effective as a nutritional supplement composition meeting the above purpose. By using the composition, the amino acid pattern in plasma and the molar ratio of branched-chain amino acid / aromatic amino acid (Fisher ratio) are corrected, hepatic encephalopathy symptoms are improved, and the nutritional status improving effect is sufficiently improved. In addition, it has been found that it is possible to maintain the nutritional status of patients with muscular dystrophy, maintain good nutrition, delay the progression of the disease state, increase the resistance to complications and sequelae, and prolong the life. Was. Furthermore, it has been found that the administration of the composition for nutritional supplementation has an effect of suppressing body protein degradation at the time of biological invasion and promoting protein synthesis. The present invention has been completed based on these findings.

That is, the present invention provides a peptide-containing substance containing a peptide having a molecular weight of 200 to 4,000 obtained by hydrolyzing a corn protein with an enzyme and having an aromatic amino acid content of 3.6 mol% or less based on all amino acids in the amino acid composition. A composition containing 5 to 20% by weight on a dry basis, wherein the content of total nitrogen compounds, carbohydrates and lipids as a protein source in the composition is 15 to 40 on a dry weight basis.
A nutritional supplement composition comprising 50% to 70% by weight of a carbohydrate and 0 to 15% by weight of a lipid, a nutritional supplement composition for a liver disease patient having the above composition, and a nutritional composition for a muscular dystrophy disease patient having the above composition. It is intended to provide a supplement composition, and a nutritional supplement composition for invasion of a living body having the above composition.

 Hereinafter, the nutritional supplement composition of the present invention will be described in detail.

Nitrogen compounds as a protein source constituting the composition of the present invention, as long as it contains a predetermined amount of a specific peptide-containing substance derived from corn protein, other known various protein raw materials,
Any of amino acids and the like may be used. As the protein raw material,
Examples thereof include salts of casein, sodium caseinate, calcium caseinate, and the like, and enzymatically decomposed products of such caseins, soybean protein, and wheat protein. As the amino acid, a branched-chain amino acid such as valine, leucine, isoleucine and the like is preferable. These nitrogen compounds are 1
They can be used alone or in combination of two or more.

The nitrogen compound as a protein source is preferably blended to improve the amino acid score from the viewpoint of nutritional effects.

When the composition of the present invention is used as a nutritional supplement composition for patients with muscular dystrophy, it is preferable to add taurine, which has been conventionally used as a therapeutic drug for muscular dystrophy.

The peptide-containing substance to be added to the composition of the present invention is as follows.

That is, the peptide-containing substance has a molecular weight of 200 to 4000 obtained by hydrolyzing corn protein with an enzyme.
Wherein the aromatic amino acid in the amino acid composition is set to 3.6 mol% or less based on all amino acids.

The method for producing a peptide-containing substance used in the present invention comprises a first step in which a raw material containing corn protein is treated with a raw starch-decomposing enzyme to decompose and remove starch, and a heat treatment of the treated product under a high alkali. And a third step of hydrolyzing with a highly alkaline protease, and a fourth step of removing aromatic amino acids.

In the above production method, a fourth step of hydrolysis using a neutral protease and / or an acidic protease is performed after the third step of hydrolysis with a highly alkaline protease, and then a fifth step of removing aromatic amino acids is performed. The process may be performed.

In the above description, branched-chain amino acid (BCAA) means leucine, isoleucine, valine and the like, and aromatic amino acid (AAA) means phenylalanine, tyrosine, tryptophan and the like.

Hereinafter, the peptide-containing substance used in the present invention will be described in more detail with reference to preferred embodiments.

In the preparation of the peptide-containing substance, as the corn protein, corn protein and / or zein (Zein), a prolamin protein that constitutes the corn protein, has a high content of branched-chain amino acids, particularly leucine, as a constituent amino acid. It is preferably used. As the corn protein, in the process of producing corn starch, corn protein suspension obtained from corn through wet milling (wet sulfurous acid immersion), for example, corn gluten meal suspension, 70% from corn gluten liquor and corn protein Zein eluted with hydrous alcohol or alkali is preferably used. When these protein suspensions are used as raw materials, the solid content concentration is 5%.
More preferably, it is adjusted to about 15% by weight.

In the method for producing a peptide-containing substance used in the present invention, as a first step, these raw materials are previously treated with a raw starch-degrading enzyme to decompose and remove starch. According to a preferred embodiment,
To the suspension of the vegetable protein, sodium hydroxide,
Potassium hydroxide, calcium hydroxide or the like is added to an alkali to adjust the pH to about 5 to 6, and a raw starch-decomposing enzyme, for example, “Daviase” (trade name, manufactured by Daikin Industries, Ltd.) is used in an amount of 0.02 to 0.2 wt% per solid material. Add 3-20 at 50-60 ° C
The reaction is performed by stirring for a period of time, followed by dehydration and filtration.

Next, as a second step, the treated product is solidified at a solid concentration of 5 to 5%.
Resuspend to 20 wt%, preferably 10-15 wt%,
An alkali such as sodium hydroxide or potassium hydroxide is added to the suspension, and the pH is preferably adjusted to 12 or more.
Heat treatment at ~ 130 ° C for 5-30 minutes. By this treatment, the vegetable protein can be denatured, and the attack by the protease can be facilitated.

Next, as a third step, the suspension is cooled to 30 to 60 ° C., more preferably to about 50 ° C., and 0.2 to 2 wt% of the high alkali protease is added to the solid content of the raw material. The reaction is preferably performed for 1 to 2 hours. By this treatment, the vegetable protein is hydrolyzed to an appropriate length.

In this case, as the highly alkaline protease, for example, an alkaliphilic bacterium (Bacillus, No. 221) reported in Horikoshi et al., "Agric. Bicl. Chem. 35 (9). 1407-1414".
Alkaline protease (from Meito Sangyo Co., Ltd.)
“Esperase 8.0L” and “Savinase 8.0L” (trade name, manufactured by Novo Corporation) derived from an alkaliphilic mutant are preferred. These highly alkaline proteases have an optimum pH for enzyme action of 10 to 12, are excellent in heat resistance, and are usually endo-type enzymes.

Next, as a fourth step, an acid such as hydrochloric acid is added to the suspension, if necessary, to obtain a neutral pH of 8 to 5.5, preferably around 7, or a pH of 5.5 to 3.0, preferably 3.5 to 3.0. Adjust to 4.0 acidity. When the pH is adjusted to 8 to 5.5, a neutral protease is added, and when the pH is adjusted to 3.5 to 4.0, an acidic protease is added in an amount of 0.2 to 2% by weight per solid content of the raw material. More preferably, the reaction is carried out at about 50 ° C. for 10 minutes to 24 hours, more preferably 20 to 24 hours. in this case,
Preferably, the neutral or acidic protease is in exo form. By this treatment, the hydrolysis rate of the vegetable protein can be further increased, and the aromatic amino acid can be converted to the free amino acid. In order to increase the molecular weight of the peptide, the fourth step may be omitted.

Incidentally, as the neutral or acidic exo-type protease, aminopeptidase and carboxypeptidase,
There are dipeptidyl peptidase and the like, and any peptidase may be used. For example, “Protease A and M” (trade name, manufactured by Amano Pharmaceutical Co., Ltd., “Sumiteam AP, MP, LP”)
(Trade name, manufactured by Shin Nippon Chemical Co., Ltd.) and the like are preferable.

Further, aromatic amino acids are removed from the enzymatic hydrolyzate of vegetable protein as a fourth step when the fourth step is omitted, and as a fifth step when the fourth step is performed. As one of the removing methods, a method using an adsorbent can be adopted. As the adsorbent, for example, activated carbon or various adsorption resins can be used. More preferably, a polystyrene-based adsorption resin such as "Dowex S-11"
Using an adsorbent obtained by activating 2 "(trade name, manufactured by Dow Chemical Co., Ltd.) with hydrochloric acid, an enzyme hydrolyzate is passed through a column filled with the adsorbent, and eluted on the acidic side. In addition, as the adsorption resin, for example, a resin disclosed in JP-A-60-136543 can be used.

Among the adsorbed substances, activated carbon has a property of adsorbing coloring substances and aromatic amino acids in a slightly acidic protein enzyme hydrolyzate. Further, the polystyrene-based adsorption resin has many benzene rings, and the benzene ring on the resin side and the benzene ring of the aromatic amino acid are hydrophobically bonded. Therefore,
By using these adsorbents, aromatic amino acids can be selectively adsorbed and removed.

In addition, with these adsorbents alone, aromatic amino acids, particularly tyrosine, are insufficiently removed, so that the low-solubility properties of tyrosine are utilized to separate and remove tyrosine by isoelectric point precipitation. Aromatic amino acids can be reduced.

The peptide-containing substance used in the present invention can be obtained through each of the above steps.When commercializing the product, the treatment solution of the last step is concentrated and the pH is adjusted, if necessary, followed by acid treatment, amidase, and deaminase. It is preferable to reduce the bitterness by an enzyme treatment, etc., desalting treatment with an ion exchange membrane or ion exchange resin, etc., further treating with activated carbon, and evaporating to dryness to obtain a powder. However, depending on the application, the solution can be used as it is.

The peptide-containing substance used in the present invention comprises a peptide obtained by hydrolyzing a vegetable protein with an enzyme as described above, and contains a peptide having a molecular weight distribution of about 200 to 4,000 and an average molecular weight of about 500 to 2,000.

The components in the peptide-containing substance are preferably composed of 50 to 92% by weight of peptide, 5 to 20% by weight of carbohydrate, 0 to 25% by weight of amino acid, and 3 to 5% by weight in solid content.

Further, the peptide-containing substance used in the present invention has an aromatic amino acid in the amino acid composition of 3.6 to all amino acids.
Mol% or less.

In a preferred embodiment, the molar ratio of branched-chain amino acid (BCAA) / aromatic amino acid (AAA) in the amino acid composition is 10 or more, more preferably 20 or more.

In a further preferred embodiment, the branched-chain amino acid is at least 10 mol% and the aromatic amino acid is at most 2 mol% in the amino acid composition.

The peptide-containing substance used in the present invention has the following amino acid composition as a preferred example.

The carbohydrates constituting the composition of the present invention may be any of various known ones other than those contained in the above-mentioned peptide-containing substance, for example, glucose, maltose, sucrose, isomaltose, maltotetraose, maltotriose, maltopentaose. Aose, maltohexaose, lactose,
Glycogen, dextrin, monosaccharides of starch sugars, oligosaccharides, dietary fiber, polysaccharides and the like can be exemplified.

As the lipid constituting the composition of the present invention, in addition to those contained in the above-mentioned peptide-containing substance, various conventionally known lipids such as rice oil, cottonseed oil, corn oil, soybean oil, sunflower oil, cocoa butter, sesame oil, Safflower oil, peanut oil, butter, lard, coconut oil, nut oil, palm oil, rapeseed oil,
Any animal or vegetable oil such as medium chain fatty acid (MCT) may be used, and vegetable oil is particularly preferred.

The composition of the present invention contains the above-mentioned peptide-containing substance in the following prescribed amounts, and it is important that the content ratio of the total nitrogen compounds, carbohydrates and lipids as the protein source be the following prescribed amounts. is there.

That is, the compounding amount of the above-mentioned peptide-containing substance is 5 to 20% by weight, preferably 10 to 15% by weight based on the dry weight of the composition of the present invention. In addition, the compounding ratio of total nitrogen compounds, carbohydrates and lipids as a protein source is 15 to 40% by weight of nitrogen compounds and 50 to 50% of carbohydrates based on the dry weight of the composition of the present invention.
The content may be 70% by weight and 0 to 15% by weight of a lipid, preferably 20 to 30% by weight of a nitrogen compound, 58 to 65% by weight of a carbohydrate, and 3 to 10% by weight of a lipid. The content of each component of the nitrogen compound, carbohydrate and lipid in the composition of the present invention is the total amount of the components contained in the peptide-containing substance and the components added to the other components.

By adjusting the composition of the present invention to the above-mentioned mixing ratio, it is possible to correct the amino acid lance in blood and to improve the nutritional state.

The composition of the present invention may further contain usual various additives as required in addition to the above three components as essential components. Examples of the additive include various vitamins (vitamin A, vitamin B ₁ , vitamin B ₂ , vitamin B ₆ ,
Vitamin B _12, vitamin C, vitamin D, vitamin E,
Niacin, folic acid, pantothenic acid, etc.), minerals (calcium, iron, potassium, sodium, magnesium,
Salts such as phosphorus and chlor), flavors such as synthetic flavors and natural flavors, sweeteners such as natural sweeteners (such as thaumatin and stevia) and synthetic sweeteners (such as saccharin and ticlo), coloring agents, emulsifiers, stabilizers, Preservatives and the like can be exemplified, and these can be used alone or in combination of two or more.

Each of the above components is usually stored or distributed in a moisture-proof bag, bottle, can, or the like in a tightly mixed powder state, if desired, but may be in the form of a beverage, jelly, granule, tablet, capsule, or the like, if desired. May be formulated.

When the nutritional supplement composition of the present invention thus obtained is applied (ingested), it is usually diluted to an appropriate concentration with water to obtain a solution or jelly form in accordance with a conventional method. This is also subjected to a treatment such as heat sterilization or pressure heat sterilization as necessary. The degree of the above dilution can be arbitrarily determined, but usually when prepared in the form of a tube or light glory solution,
Suitably, the resulting nutrient solution contains about 100 to 500 g / preferably about 150 to 250 g / of the composition of the present invention. The nutrient solution is administered orally or directly by tube to the stomach, duodenum or small intestine, and the jelly preparation is orally administered. The amount of administration (ingestion) can be appropriately determined depending on the disease state of the patient to be administered and the intended treatment or nutritional improvement effect, etc., and is generally calculated as the dry weight of the composition of the present invention per person per day.
It is desirable to be about 30 to 150 g.

The composition of the present invention can be used in combination with a normal diet in consideration of patient's taste.

Effect of the Invention The nutritional supplement composition of the present invention, which mainly contains a peptide as a protein source, has a good taste, has no unpleasant bitterness, has a rapid and good absorption from the digestive tract, and has an osmotic pressure. The occurrence of diarrhea due to being high is avoided. Further, the compounded peptide-containing substance has an amino acid composition of:
It contains abundant branched-chain amino acids derived from plant proteins, and has a very small amount of aromatic amino acids, and can significantly improve the amino acid pattern and the Fisher ratio in plasma of liver disease, Furthermore, since the protein source, carbohydrate and fat are contained in a well-balanced manner, the nutritional status can be improved. For this reason, the nutritional supplement composition of the present invention is suitable as a dietary supplement for hepatitis recovery period, chronic hepatitis or liver cirrhosis, especially as a nutritional supplement or therapeutic diet for liver disease patients such as fulminant hepatitis and chronic hepatic encephalopathy. It is something.

In addition, even in patients with muscular dystrophy, the intake of the composition of the present invention corrects the amino acid imbalance in the blood, and can further maintain the nutritional status of the patient,
It can delay the progression of the condition, increase resistance to complications and sequelae, and prolong life.

Furthermore, the administration of the composition of the present invention exerts the effect of suppressing body protein degradation and promoting protein synthesis at the time of invasion of a living body, and can maintain a good nutritional state. It is suitable.

EXAMPLES Hereinafter, the present invention will be described in more detail with reference to Reference Examples, Examples, and Test Examples.

REFERENCE EXAMPLE 1 Potassium hydroxide was added to corn protein suspension (gluten liquor) 450 obtained from the wet milling process to adjust the pH to 5.5, and "Dabiase" (trade name, Daikin Industries, Ltd.) was used as a raw starch-degrading enzyme. ) Made 70g
Was added and reacted at 50 ° C. for 16 hours under stirring. The suspension was subjected to solid-liquid separation using a filter press, and the water-soluble fraction was removed to obtain 80 kg of a corn protein wet cake. Re-suspend this wet cake in distilled water 350,
Under stirring, the temperature was raised to 80 ° C. Next, the pH was adjusted to 12.0 by adding potassium hydroxide, and heat treatment was performed at 125 ° C. for 5 minutes.

Next, the suspension was cooled to 50 ° C., and 80 g of a highly alkaline protease (manufactured by Meito Sangyo Co., Ltd.) derived from an alkalophilic bacterium (Bacillus. No. 221) was added and reacted for 1.5 hours. Furthermore, hydrochloric acid was added to the suspension to adjust the pH to 7.0, and 180 g of "Denazyme AP" (trade name, manufactured by Nagase Biochemical Co., Ltd.), a neutral protease, was added, and reacted at 50 ° C for 20 hours. I let it. The reaction solution is separated into solid and liquid by a filter press,
The water-soluble fraction was concentrated to obtain Bx20, 160 corn protein enzyme hydrolysis.

A part of the hydrolyzate was adjusted to pH 4.0 with hydrochloric acid, and the polystyrene-based adsorption resin “Dowex S-112” (trade name: manufactured by Dow Chemical Co., Ltd.) was activated with hydrochloric acid. The hydrolyzate was passed through a column to separate a low fraction of aromatic amino acids. After concentrating the passing solution, desalting is performed with an ion-exchange membrane (micro-acylizer: manufactured by Asahi Kasei Corporation), treated with activated carbon, lyophilized, and then subjected to total nitrogen 14.30.
%, Amino nitrogen 3.00%, sugar content 5.20%, ash content 1.0%, and a white powder having a branched chain amino acid to aromatic amino acid molar ratio of 38.3 was obtained. The amino acid composition is shown in Table 1.

Reference Example 2 The neutral protease “denazyme” in Reference Example 1 above
AP "(trade name, manufactured by Nagase Seikagaku Co., Ltd.) was replaced with an acidic protease" protease Amano M "(trade name, manufactured by Amano Pharmaceutical Co., Ltd.), except that the reaction pH was changed to 4. The same operation as in Reference Example 1 was repeated, freeze-dried,
A white powder having a composition of 13.50% of total nitrogen, 2.95% of amino nitrogen, 5.00% of sugar, and 1.2% of ash and having a molar ratio of branched-chain amino acid to aromatic amino acid of 69.1 was obtained. The amino acid composition is shown in Table 1.

Reference Example 3 The same operation as in Reference Example 2 was performed, passing through the adsorption resin,
The fraction having a lower aromatic amino acid content was separated, the pH was adjusted to 5.5 with potassium hydroxide, and concentrated to a solid concentration of 15% (Bx15). After the concentrate was cooled at 5 ° C., the precipitated tyrosine was removed by filtration. Thereafter, desalting, activated carbon treatment, and lyophilization were performed in the same manner as in Reference Example 2 to obtain 13.8% of total nitrogen and 2.50 of amino nitrogen.
%, A sugar content of 4.5%, and an ash content of 1.3%, and a white powder having a molar ratio of branched-chain amino acids to aromatic amino acids of 28.5 was obtained. The amino acid composition is shown in Table 1.

Reference Example 4 The same operation as in Reference Example 2 was performed, and the fraction from which tyrosine was removed by filtration in Reference Example 3 was desalted, treated with activated carbon, and then concentrated to Bx60. The cloudy concentrate is separated into a precipitate section and a soluble section by centrifugation, and the precipitate section is directly dried under reduced pressure, and the total nitrogen is 13.5%, the amino nitrogen is 2.92%, the sugar content is 4.8%, and the ash content is 1.1%.
And a molar ratio of branched-chain amino acids to aromatic amino acids of 46.1 was obtained. The amino acid composition is shown in Table 1.

Reference Example 5 The same operation as in Reference Example 3 was performed, and the excess obtained by filtering and removing tyrosine was passed through a cation exchange resin and an anion exchange resin. The elution fraction was separated, and the pH was adjusted to 4.0 with potassium hydroxide. Thereafter, desalting, activated carbon treatment and freeze-drying were performed in the same manner as in Reference Example 1 to obtain a total nitrogen of 14.0%, amino nitrogen of 0.90%, and a sugar content of 5.
A white powder having a composition of 0% and an ash content of 1.0% and having a molar ratio of the branched-chain amino acid to the aromatic amino acid of 25.6 was obtained. The amino acid composition is shown in Table 1.

Reference Example 6 The same operation was performed up to the enzymatic reaction using the highly alkaline prosthesis in Reference Example 1, and hydrochloric acid was added to the reaction solution to adjust the pH.
Adjusted to 5.5. Apply the reaction solution to a filter press,
After solid-liquid separation, the water-soluble fraction was concentrated to Bx15 to obtain 230 corn protein enzyme hydrolysates.

Next, the fraction passed through the adsorption resin was passed through a cation exchange resin and an anion exchange resin in the same manner as in Reference Example 1, the elution fraction was separated, the pH was adjusted to 5.5 with potassium hydroxide, and desalination was performed. Activated carbon treatment, freeze-dried, consisting of 14.1% total nitrogen, 0.55% amino nitrogen, 4.8% sugar, 0.9% ash,
The molar ratio of branched-chain amino acids to aromatic amino acids is 21.1
Was obtained. The amino acid composition is shown in Table 1.

Table 1 does not include data on tryptophan, which is a kind of aromatic amino acid, because tryptophan was scarcely contained.

From the results in Table 1, the enzymatic hydrolysates obtained in Reference Examples 1 to 6 have a remarkable ratio of the branched amino acids (leucine, isoleucine, valine) to the aromatic amino acids (phenylalanine, tyrosine) in the amino acid composition. You can see that it is higher. In Reference Examples 3 to 6,
It can be seen that branched chain amino acids are contained in a ratio equal to or higher than the starting material. In addition, since the total nitrogen is 14 and the protein content is high, and the amino nitrogen is 3.0 or less, it can be understood that the peptide is rich in peptides.

Reference Example 7 The peptide compositions obtained in Reference Examples 1 and 3 were analyzed by high performance liquid chromatography (HPLC).
The molecular weight distribution of each of the peptide compositions was determined.

"OH Pak KB-802.5" (trade name,
Flow rate of 0.5 ml / mi using a 0-1% trifluoroacetic acid-40% acetonitrile eluent using Showa Denko KK
The analysis was performed with n as the detector and UV 210 nm as the detector. Also,
As standard samples, tyrochrome C (MW ≒ 10.000), oligopeptides Gly-Gly-Gly (MW ≒ 189.2), Gly-G
ly (MW ≒ 132) and tyrosine (MW
≒ 181.2) and phenylalanine (MW 5.2165.2).

The analysis results of the peptide composition of Reference Example 1 are shown in FIG. 1, and the analysis results of the peptide composition of Reference Example 3 are shown in FIG. In the figure, the elution positions of the standard samples are indicated by arrows.

In FIGS. 1 and 2, complete separation and identification of the free amino acid and the peptide are not performed, but in consideration of the fact that the minimum molecular weight of the peptide is usually around 200, Reference Example 1 was used.
And the molecular weight of the peptide contained in the peptide composition of 3.
It turns out that it is in the range of 200-4000.

Example 1 The nutritional supplement composition of the present invention was prepared by mixing the components using the peptide-containing substance obtained in the above Reference Example so that the composition (g) shown in Table 2 below per 100 g of dry weight. .

Test Examples The following test examples were used to examine the effect of the peptide-containing substance contained in the composition of the present invention on improving the amino acid pattern, Fisher ratio, and the like.

Test Example-I As a peptide-containing substance, a substance prepared in the same manner as in Reference Example 1 using corn gluten as a raw material was used. The amino acid composition and the amounts of other components are as shown in Table 3.

Six-week-old male SD rats were treated with the BCAA peptide I administration group (28
Animals: body weight 216.4 ± 9.1 g) and BCAA peptide II administration group (1
The experiment was conducted by dividing the animals into two groups of 9 animals: body weight 218.4 ± 9.8 g). After fasting the rats for 18 hours, the test substance was orally administered. The test substance was 1.35 g / kg in protein amount, dissolved in 10 ml of ion-exchanged water, and administered using a catheter. Before administration, and at 30, 60, 120, 180, 240, and 360 minutes after administration, blood was collected from the lower aorta under ether anesthesia. The blood was immediately centrifuged and the plasma was stored at -80 ° C.

The amino acid analysis sample was prepared by mixing 200 μl of plasma with an equal amount of 6% sulfosalicylic acid, centrifuging at 3000 rpm for 10 minutes, and removing the supernatant.
Those frozen day and night were thawed, centrifuged again, and the supernatant was used. As a coloring reagent, a ninhydrin reagent L-8500 set manufactured by Wako Pure Chemical Industries, Ltd. was used. As an elution reagent, MCI BUFFER L-8500-PF kit manufactured by Mitsubishi Kasei Kogyo Co., Ltd. was used. Amino acid analyzer is L-85 manufactured by Hitachi, Ltd.
A Hitachi custom ion exchange resin (# 2622SC) was packed in a 4.6 mm ID × 60 mm column using a type 00 high-speed amino acid analyzer. The results for amino acids in blood are shown in FIG.

In the tables, amino acids conform to the rules of IUPAC and IUB or conventional symbols in the art, and examples thereof are shown below. In addition, when there is an optical isomer with respect to an amino acid or the like, the L-form is indicated unless otherwise specified.

Leu: Leucine, Ile: Isoleucine, Ala: Alanine, Gl
u: glutamine, Thr: threonine, His: histidine, Ser:
Serine, Gly: glycine, Asn: asparagine, Arg: arginine, Asp: aspartic acid, Pro: proline.

Test Example II Anesthesia for inferior portal vena cava (PCS) in 6-week-old male SD rats
Chronic liver failure model rats (group A, 25)
And 6-week-old male SD rats (B)
Group, 25 animals) for 2 weeks.

For these rats, changes in plasma free amino acid concentration and brain free amino acid concentration after a single administration of the peptide-containing substance were examined by the following method. The administered peptide-containing substances are as shown in Table 4 below.

After breeding rats in each group, 5 animals each in groups A and B were sacrificed, and the peptide-containing substance was administered once to each of the remaining groups in a single dose of 1.35 g / kg. At 15, 30, 60, and 120 minutes after administration, 5 animals per group were sacrificed, and the plasma free amino acid concentration and the free amino acid concentration in the whole brain were measured in the same manner as in Test Example I. The results are shown in FIGS. 4 and 5.

From these results, when a peptide-containing substance derived from corn gluten was evaluated, the total amino acid concentration, BCAA concentration and E / N ratio in the blood were increased, and the peptide-containing substance had a high nutritional supplement effect, It was found that the amino acid pattern, the Fisher ratio, etc. in the medium could be improved. Therefore, the composition of the present invention, in which a nitrogen compound as a protein source, carbohydrate and lipid are blended in a well-balanced manner, has a high nutritional supplementing effect, and is particularly useful for nutritional supplementation at the time of liver disease patients, muscular dystrophy patients, and biological invasion. It turns out to be something.

[Brief description of the drawings]

FIG. 1 is a chart showing an elution curve when the peptide composition obtained in Reference Example 1 was subjected to high performance liquid chromatography. FIG. 2 is a table showing the peptide composition obtained in Reference Example 3 subjected to high performance liquid chromatography. FIG. 3 is a graph showing a time-dependent change in blood amino acid concentration in Test Example I, and FIG.
FIG. 5 is a graph showing the temporal change of the BCAA concentration, and FIG. 5 is a graph showing the temporal change of the BCAA / AAA ratio in Test Example II.

──────────────────────────────────────────────────続 き Continuation of front page (72) Inventor Hirofumi Ueno 3-11-21 Nagatoishi Nagatoishi, Kurume City, Fukuoka Prefecture No. 406, Residence Denso Ranhou 406 (56) References JP-A-2-300137 (JP, A) JP-A-2 -138991 (JP, A) (58) Fields investigated (Int. Cl. ⁶ , DB name) A23L 1/305 A61K 37/18 A23L 2/00-2/38

Claims (8)

(57) [Claims] 1. A peptide-containing substance containing a peptide having a molecular weight of 200 to 4000 obtained by hydrolyzing a corn protein with an enzyme and having an aromatic amino acid content of 3.6 mol% or less based on all amino acids in the amino acid composition. A composition containing 5 to 20% by weight on a dry basis, wherein the total content of nitrogen compounds, carbohydrates and lipids as a protein source in the composition is 15 to 40% by weight of a nitrogen compound and 50% of carbohydrate on a dry weight basis. A nutritional supplement composition, characterized in that it is from about 70% by weight and from 0 to 15% by weight of lipids. 2. The composition according to claim 1, which is a nutritional supplement composition for patients with liver disease. 3. The composition according to claim 1, which is a nutritional supplement composition for patients with muscular dystrophy. 4. The composition according to claim 1, which is a nutritional supplement composition for invasion of a living body. 5. The composition according to claim 1, wherein the molar ratio of branched-chain amino acid / aromatic amino acid is 10 or more in the amino acid composition of the peptide-containing substance. 6. The composition according to claim 1, wherein in the amino acid composition of the peptide-containing substance, the content of the branched-chain amino acid is 10 mol% or more and the content of the aromatic amino acid is 2 mol% or less. 7. The composition according to claim 1, wherein the peptide-containing substance has the following amino acid composition. 8. The method according to claim 8, wherein the peptide-containing substance is contained in a solid content of the peptide.
50-92% by weight, carbohydrates 5-20% by weight, amino acids 0-25
The composition according to any one of claims 1 to 3, which further comprises 3 to 5% by weight.