JPH03251153A - Nutrition supplying composition - Google Patents

Abstract

PURPOSE:To obtain the title composition suitable as nutrition supplying food for patients of hepatic disease, etc., treating food, etc., comprising a given amount of a peptide-containing substance having a specific composition prepared by hydrolyzing maize protein with an anzyme, by controlling the blended amount of each of nitrogen compound, carbohydrate and lipid. CONSTITUTION:Maize protein is hydrolyzed with an enzyme (e.g. highly alkali- side protease) to produce a peptide-containing substance which contains peptide having 200-4,000molecular weight and has an amino acid composition wherein an amount of aromatic amino acid is <=3.6mol% based on total amounts of amino acids and a molar ratio of branched-chain amino acid/aromatic amino acid is preferably >=10. Then 5-20wt.% based on dry weight of the peptide- containing substance is blended with a nitrogen compound, a carbohydrate and a lipid so as to give 15-40wt.% nitrogen compound, 50-70wt.% carbohydrate and 0-15wt.% lipid to produce a nutrition supplying composition.

Description

DETAILED DESCRIPTION OF THE INVENTION Field of the Invention The present invention relates to a nutritional supplement composition, and more specifically, it is particularly suitable as a nutritional supplement or therapeutic food for liver disease patients, muscular dystrophy patients, and during biological invasion. The present invention relates to nutritional supplement compositions.

Conventional techniques and their challenges In patients with chronic liver failure such as cirrhosis, toxic substances generated in the body due to the formation of intrahepatic and extrahepatic shunts and liver dysfunction bypass the liver parenchyma, making it difficult to detoxify the body, and also impairing substance metabolism. Combined with other abnormalities, it is said that this can eventually lead to metabolic abnormalities in the brain, making it easy for the patient to fall into a coma.

In 1974, Fisher measured plasma amino acid concentrations in patients with severe liver disease and found that in patients with hepatic coma due to liver cirrhosis, there was a marked increase in aromatic amino acids (AAA) such as phenylalanine, tyrosine, and free tryptophan, as well as methionine and valine. A decrease in branched chain amino acids (BCAAs) such as , leucine, and isoleucine was observed.
It was reported that amino acid metabolic abnormalities are the main cause of hepatic encephalopathy. In order to improve hepatic encephalopathy by correcting the amino acid imbalance in plasma caused by abnormal amino acid metabolism, we have developed an amino acid infusion that limits aromatic amino acids and increases the content of branched chain amino acids. (
Fisher's solution) was developed. This amino acid infusion has an excellent awakening effect on patients with hepatic encephalopathy, so its usefulness is highly evaluated and it is widely applied clinically.

For patients with liver cirrhosis who have recovered from hepatic encephalopathy, it is desirable to prevent the recurrence of hepatic encephalopathy and improve their nutritional status, but even during the period when they exhibit hypoalbuminemia, they are forced to undergo protein-restrictive diet therapy that focuses on the former. However, with such conventional therapy, it is not possible to correct the amino acid imbalance in plasma that leads to hepatic encephalopathy, nor can it be expected to improve nutritional status.
It does not reach the point of increasing the patient's own physical activity. Furthermore,
If protein intake is restricted for a long period of time, hypoalbuminemia will become worse.

In the case of liver cirrhosis patients, a high-calorie, high-protein, high-vitamin diet is considered desirable in order to actively recover. Therefore, in patients with liver cirrhosis who have symptoms of hepatic encephalopathy or are at risk of developing symptoms, not only treatment or prevention of hepatic encephalopathy but also nutritional considerations are extremely important in patient management. If nutritional management is inadequate, the patient is always at risk of developing hepatic encephalopathy. In this sense, there are natural limitations to the management of hepatic encephalopathy using infusions, and there has been a strong desire to develop a formulation that contributes to both improvement of hepatic encephalopathy symptoms and nutritional status for maintenance therapy after hepatic encephalopathy awakens.

From this perspective, in order to correct the amino acid imbalance in the plasma and improve the nutritional status of patients with liver disease, Fisher's theory Nutrient supplements with high amino acid content and various nutrients are being considered. However, such nutritional supplements use amino acids as a protein source, which has the disadvantage of high osmotic pressure.
Diarrhea has been observed when ingested orally. Furthermore, amino acids have a unique odor and bitterness, which also poses problems.

On the other hand, muscular dystrophy (PMD) is a group of diseases that have a primary origin in the muscles themselves, and is mainly characterized by progressive degeneration of skeletal muscles, and is a disease that develops on a genetic basis. Examining the amino acid composition in the blood of muscular dystrophy patients, even when protein intake sufficiency is not necessarily poor, the ratio of essential amino acids to non-essential amino acids (E/N ratio), which is an indicator of malnutrition, is reduced. An abnormality in protein amino acid metabolism is considered. In addition, it has been reported that the concentration of branched chain amino acids in the blood is lower than that of normal humans (Kumiko Hirano, Bulletin of the Faculty of Life Sciences, Osaka University, 34; 253-25.9.1986), and this report This suggests the need to increase the concentration of branched chain amino acids in the blood.

Furthermore, there is a report that branched chain amino acid supplementation has the effect of suppressing muscle decline in patients with amyotrophic lateral sclerosis (ALS), a type of muscular dystrophy.
Plailakig A, ej al :Lan
ce+, May 7; 1015-1018.1988
).

In order to prolong the lives of patients with muscular dystrophy, it is necessary to provide sufficient energy and protein to slow down weight loss, but it is not easy to increase the amount of food eaten by patients with occlusal disorders. High-energy, high-protein supplementation is required at times.

In addition, during times of stress, especially surgical invasion, wounds, burns,
A typical metabolic change during biological invasion, such as during sepsis, is increased catabolism of protein metabolism. In other words, body proteins such as skeletal muscle, visceral smooth muscle, and subcutaneous collagen tissue are broken down and consumed as an energy source, while at the same time acute-phase proteins (fibrinogen, haptoglobin,
Although it is reused for the biosynthesis of lysosomal enzymes, etc., it is generally used for proteolysis. As a result, urinary nitrogen excretion increases, nitrogen balance takes on a negative value, leading to weight loss and weakening recovery ability.

Goldberg (A, L, Goldberg) et al.
AA), a series of in-house tests using rat diaphragm muscle.
In the vNro experiment, we reported for the first time that BCAA has the effect of inhibiting muscle protein breakdown or promoting synthesis.

Blackburn (G, L, B Iackbur
n) Raya Fische'r - (J, E, Fische
Since the end of the 1970s, using various invasive animal models, they have shown that protein metabolism during stress can be improved by administering an amino acid infusion with a high BCAA content.

Based on the results of these tests, in the 1980s B
An amino acid infusion containing a high concentration of CAA was prototyped and tested in the clinic, and clinically it was confirmed that it has a significant nutritional effect under stress.

Based on these findings, it is desired to develop nutritional foods with a high content of branched chain amino acids for patients with muscular dystrophy or for nutritional supplementation during biological invasion. Nutrient supplements have problems such as excessively high osmotic pressure and the unique odor and bitterness of amino acids, making them unsuitable as oral nutritional supplements for humans.

The present invention has been made in view of the above-mentioned problems of the prior art, and its purpose is to enable oral and enteral administration such as through a tube, to achieve physiological digestion and absorption, and to have a high osmotic pressure. This is a nutritional supplement composition that can avoid the occurrence of diarrhea caused by food poisoning, and has no irritation or foreign taste, and that by ingesting the composition, amino acid imbalance in the blood can be corrected, and the nutritional status can be improved. It is an object of the present invention to provide a nutritional composition that can be used as a nutritional supplement.

Means for Solving the Problems The present inventor has conducted extensive research in order to achieve the above object. As a result, through specific processing, raw materials containing corn protein can be produced with a molecular weight of 200 to 400.
We have discovered that it is possible to obtain a peptide-containing substance that has peptide 0 as its main component, contains abundant branched-chain amino acids, and contains almost no aromatic amino acids. It has been found that a composition in which the amounts of total nitrogen compounds, carbohydrates, and lipids as protein sources are adjusted to a predetermined ratio is effective as a nutritional supplement that meets the above objectives. By using this composition, the amino acid pattern and the molar ratio of branched chain amino acids/aromatic amino acids (Fisher ratio) in plasma are corrected, and symptoms of hepatic encephalopathy are improved, and the effect of improving nutritional status is also sufficient. Furthermore, we have discovered that it is possible to maintain a good nutritional status in patients with muscular dystrophy, delay the progression of the disease, increase resistance to complications and sequelae, and prolong life. . Furthermore, it has been found that administration of the nutritional composition described above has the effect of suppressing body protein decomposition and promoting protein synthesis during biological invasion. The present invention was completed based on these findings.

That is, the present invention provides a peptide-containing substance that contains a peptide with a molecular weight of 200 to 4,000 obtained by hydrolyzing corn protein with an enzyme, and in which aromatic amino acids account for 3.6 mol% or less based on the total amino acids in the amino acid composition. A composition containing 5 to 20% by weight on a dry weight basis, wherein the content of total nitrogen compounds, carbohydrates and lipids as a protein source in the composition is 15 to 40% by weight on a dry weight basis, A nutritional composition characterized by containing 50 to 70% by weight of carbohydrates and 0 to 15% by weight of lipids; A nutritional composition for liver disease patients having the above composition; A nutritional composition for muscular dystrophy disease patients having the above composition. , and a nutritional supplement composition for biological invasion having the above composition.

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

The nitrogen compound as a protein source constituting the composition of the present invention is
As long as it contains a predetermined amount of a specific peptide-containing substance derived from corn protein, the rest may be any of various known protein raw materials, amino acids, etc.

Examples of the protein raw material include casein and salts such as sodium caseinate and calcium caseinate, enzymatic decomposition products of secondary caseins, soybean protein, and wheat protein enzymatic decomposition products. Furthermore, as the amino acid, branched chain amino acids such as valine, leucine, and isoleucine are preferred. These nitrogen compounds can be used singly or in combination of two or more.

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

Furthermore, when the composition of the present invention is used as a nutritional supplement composition for patients with muscular dystrophy, it is preferable to incorporate taurine, which has been conventionally used as a therapeutic agent for muscular dystrophy.

The peptide-containing substances to be incorporated into the composition of the present invention are explained below.

That is, the peptide-containing substance has a molecular weight of 200 to 40 and is obtained by hydrolyzing corn protein with an enzyme.
00 peptide, and is characterized in that its amino acid composition contains 3.6 mol% or less of aromatic amino acids based on the total amino acids.

Furthermore, the method for producing the peptide-containing substance used in the present invention is as follows:
The first step is to treat the raw material containing corn protein with a raw starch-degrading enzyme to decompose and remove starch, the second step is to heat-treat this treated product under high alkaline conditions, and the second step is to hydrolyze it with a high alkaline protease. It is characterized by comprising three steps and a fourth step of removing aromatic amino acids.

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

In addition, in the above, branched chain amino acids (BCAA) are
Aromatic amino acids (AAA) include leucine, isoleucine, valine, etc., and aromatic amino acids (AAA) include phenylalanine, tyrosine,
It means tryptophan etc.

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

In the preparation of the above-mentioned peptide-containing substance, the corn protein used is corn protein and/or Zein, a prolamin protein that constitutes corn protein, because it has a high content of branched chain amino acids, especially leucine, as a constituent amino acid. Preferably used. This corn protein includes a corn protein suspension obtained from corn through wet milling (wet sulfite immersion) in the corn starch manufacturing process, such as a corn gluten meal suspension, corn gluten liquor, and 70% corn protein. Zein, which is eluted with hydrous alcohol or alkali, is preferably used. When these protein suspensions are used as raw materials, it is more preferable to adjust the solid content concentration to about 5 to 15%.

In the method for producing a peptide-containing substance used in the present invention, the first
As a step, these raw materials are treated in advance with a raw starch-degrading enzyme to decompose and remove starch. According to a preferred embodiment, an alkali such as sodium hydroxide, potassium hydroxide, or calcium hydroxide is added to the suspension of the vegetable protein to adjust the pH to about 5 to 6, and a raw starch degrading enzyme, such as "Dabiase" (trade name, manufactured by Daikin Industries) was added in an amount of 0.02 to 0.2 wt% based on the solid content of the raw material, and
The reaction is carried out by stirring at 0° C. for 3 to 20 hours, followed by dehydration and filtration.

Next, as a second step, this treated product is mixed with a solid content of 5 to 2
Resuspend to 0wt%, preferably 10 to 15W1%, add an alkali such as sodium hydroxide or potassium hydroxide to this suspension, adjust the pH to preferably 12 or higher, and heat to 100 to 130°C. Heat-process for 5 to 30 minutes. This treatment denatures vegetable proteins,
It can make it easier to attack by protease.

Next, as a third step, the suspension is cooled to 30 to 60°C, more preferably around 50°C, and 0.2 to 2w1% of high alkaline protease is added based on the solid content of the raw material.
The reaction is allowed to proceed for 0 minutes to 24 hours, more preferably for 1 to 2 hours. This treatment hydrolyzes vegetable proteins into appropriate lengths.

In this case, as the high alkaline protease, for example, Horiori et al., "Agric, Biol, Chem, 35 (9
) 1407-1414J, alkaline protease (manufactured by Meito Sangyo ■) derived from alkalophilic bacteria (Bacillus, 221), and ``Esperase 8.OLJ'' and ``Savinase 8.0L'' derived from alkaliphilic mutant strains.
J (trade name, manufactured by Novo), etc. are suitable. These highly alkaline proteases have an optimal pH for enzymatic action of 1.
0 to 12, has excellent heat resistance, and is usually an endo-type enzyme.

Next, as a fourth step, an acid such as hydrochloric acid is added to this suspension as necessary to obtain a pI (8 to 5.5, preferably 7).
Before and after neutrality, or p) ('Adjust to acidity of 3.5 to 3.0, preferably 3.5 to 4.0. Then, pH 8 to 5
．． 5, use neutral protease at pH 3.5-4.
．． If it is set to 0, add acidic protease at 0.2 to 2 wj% based on the solid content of the raw material, and adjust the optimum temperature of the enzyme, e.g.
0 to 60°C, more preferably around 50°C, for 10 minutes to 2
The reaction is allowed to proceed for 4 hours, preferably for 20 to 24 hours. In this case, the neutral or acidic protease is preferably of exo type. This treatment can further increase the hydrolysis rate of vegetable proteins and convert aromatic amino acids into free amino acids. Furthermore, in order to increase the molecular weight of the peptide, this fourth step may be omitted.

The neutral or acidic exo-type proteases include aminopeptidase, carboxypeptidase, dipeptidyl peptidase, etc. Any peptidase may be used, but for example, "Protease A and M" (trade name, manufactured by Ohno Pharmaceutical Co., Ltd., “Sumi Team AP, MP, LPJ
(trade name, manufactured by Shin Nihon Kagaku Kogyo ■) etc. are suitable.

Furthermore, aromatic amino acids are removed from the enzymatic hydrolyzate of vegetable protein in a fourth step if the fourth step is omitted, or in a fifth step if the fourth step is performed. As one method for removing it, a method using an adsorbent can be adopted. As the adsorbent, activated carbon and various adsorption resins can be used, for example. More preferably, polystyrene-based adsorption resins, such as [DOWEX S-112
Using an adsorbent made of J (trade name, manufactured by Dow Chemical ■) activated with hydrochloric acid, the enzymatic hydrolyzate is passed through a column filled with this adsorbent and eluted on the acidic side. In addition, as the adsorption resin, for example, a resin disclosed in JP-A-60-136543 can also be used.

Among the above-mentioned adsorption substances, activated carbon has the property of adsorbing colored substances and aromatic amino acids in slightly acidic protein enzyme hydrolysates. Moreover, 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 form a hydrophobic bond. Therefore, by using these adsorbents, aromatic amino acids can be selectively adsorbed and removed.

In addition, since these adsorbents alone are insufficient to remove aromatic amino acids, especially tyrosine, tyrosine can be separated and removed by utilizing the low solubility properties of 1,000 rosin by isoelectric focusing method, etc. Furthermore, aromatic amino acids can be reduced.

The peptide-containing substance used in the present invention can be obtained through each of the above steps, but when commercializing the product, if necessary, after concentrating the treatment solution in the last step and adjusting the pH, acid treatment, amidase, deaminase etc. It is preferable to reduce the bitter taste by enzymatic treatment such as the like, desalinate with an ion exchange membrane or ion exchange resin, further treat with activated carbon, and evaporate to dryness to powder. However, depending on the application, it can also be used as a solution.

The peptide-containing substance used in the present invention is made of vegetable protein hydrolyzed with an enzyme as described above, and has a total molecular distribution of 200 to 4000 and an average molecular weight of 500 to 2000.
Contains some peptides.

Preferably, the components in this peptide-containing substance include:
Peptide 50-92% by weight, carbohydrate 5-20% in solid content
Weight%, amino acids 0-25% by weight, other 3-5% by weight
It consists of

Furthermore, in the amino acid composition of the peptide-containing substance used in the present invention, aromatic amino acids account for 3.6% of total amino acids.
It is considered to be less than mol%.

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

In a more preferred embodiment, the amino acid composition includes 10 mol% or more of branched chain amino acids and 2 mol% or less of aromatic amino acids.

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

Amino acid composition (mol%) Aspartic acid 3.0-8.7 Threonine 2.1-6.2 Serine 3.4-9.6 Glutamic acid 10.9-30.4 Glycine
1.9-4.5 Alanine 8.0-26.4 Valine
3.5-6.2 Cysteine 0. 0-0.1 methionine
0.4-6.5 Isoleucine 0.9-6.5 Leucine 4.4-47.5 Tyrosine
0.1-2.0 Phenylalanine 0.0-1.6 Lysine 1.3-2.8 Histidine 1.0-2.5 Arginine 1.3-2.1 Proline 2.9-6.0 Composition of the present invention In addition to those contained in the above-mentioned peptide-containing substances, the carbohydrate constituting the product may be any of various known carbohydrates, such as glucose, maltose, sucrose, isomaltose, maltotetraose, maltotriose, maltopentaose, and maltotriose. Examples include monosaccharides such as hexaose, lactose, glycogen, dextrin, and starch, oligosaccharides, dietary fiber, and polysaccharides.

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

The composition of the present invention contains the above-mentioned peptide-containing substances in the following predetermined amounts, and it is important that the content ratios of total nitrogen compounds, carbohydrates, and lipids as protein sources be in the following predetermined amounts. be.

That is, the amount of the peptide-containing substance blended is 5 to 20% by weight, preferably 10 to 1% by weight, based on the dry weight of the composition of the present invention.
The content is preferably 5% by weight. In addition, the blending ratio of total nitrogen compounds, carbohydrates, and lipids as a protein source is 15 to 40% by weight of nitrogen compounds, 50 to 70% by weight of carbohydrates, and 0 to 15% by weight of lipids, based on the dry weight of the composition of the present invention. Preferably, the content may be 20 to 30% by weight of nitrogen compounds, 58 to 65% by weight of carbohydrates, and 3 to 10% by weight of lipids. The content of each component of nitrogen compounds, carbohydrates, and lipids in the composition of the present invention is the total amount of the components contained in the peptide-containing substance and other components added.

By setting the composition of the present invention at the above-mentioned blending ratio,
It can correct the amino acid balance in the blood and improve nutritional status.

In addition to containing the above-mentioned three components as essential components, the composition of the present invention may further contain various conventional additives as required. Examples of such additives include various vitamins (
Vitamin A1 Vitamin B1, Vitamin B2, Vitamin B
6. Vitamin B1□, vitamin C1 vitamin D1 vitamin E1 niacin, folic acid, pantothenic acid, etc.), minerals (salts such as calcium, iron, potassium, sodium, magnesium, phosphorus, chlorine, etc.), synthetic fragrances and natural fragrances, etc. Flavorings, natural sweeteners (thaumatin, stevia, etc.)
and sweeteners such as synthetic sweeteners (saccharin, chikuguchi, etc.),
Examples include colorants, emulsifiers, stabilizers, preservatives, etc., and each of these can be used singly or in combination of two or more.

Each of the above ingredients is normally stored or distributed in a tightly mixed powder state in a moisture-proof bag, bottle, can, etc., but if desired, it can be in the form of a drink, jelly, granule, tablet, capsule, etc. It may be formulated into a formulation.

When applying (ingesting) the nutritional composition of the present invention thus obtained, it is usually diluted with water to an appropriate concentration according to a conventional method to form a solution or jelly. This is also subjected to treatment such as heat sterilization or pressure heat sterilization as necessary.

The degree of dilution mentioned above can be determined arbitrarily, but when the nutritional solution is prepared in the form of a tube or oral nutritional solution, the obtained nutritional solution contains about 100 to 500 g/(1, preferably about 15
It is suitable that the content is 0 to 250 g/Ω.

The above nutritional solution is administered orally or directly into the stomach, duodenum or small intestine, and the jelly preparation is administered orally. The amount to be administered (intake) can be determined as appropriate depending on the disease state of the patient to whom it is administered, the intended treatment or nutritional improvement effect, etc.
-Generally, 30 kg of dry weight of the composition of the invention per person per day.
It is desirable to set it to about 150 g.

The composition of the present invention can also be used in conjunction with regular meals, taking into account the patient's preferences.

Effects of the Invention The nutritional supplement composition of the present invention mainly contains peptides as a protein source, has a good flavor, has no unpleasant bitterness, has rapid and good absorption from the gastrointestinal tract, and has a low osmotic pressure. In addition, the peptide-containing substance to be blended contains abundant branched chain amino acids derived from vegetable proteins in its amino acid composition, and has a very high amount of aromatic amino acids. There are few
It can significantly improve the amino acid pattern and Fischer's ratio in the plasma of liver disease patients, and furthermore, since it contains protein sources, carbohydrates, and fats in a well-balanced manner, it is possible to improve the nutritional status of patients with liver disease. Therefore, the nutritional supplement composition of the present invention is suitable as a therapeutic food for the hepatitis convalescence period, chronic hepatitis or liver cirrhosis, and especially as a nutritional supplement or therapeutic food for patients with liver diseases such as fulminant hepatitis and chronic hepatic encephalopathy. It is something.

Furthermore, in patients with muscular dystrophy, the amino acid imbalance in the blood can be corrected by ingesting the composition of the present invention, and furthermore, the patient's nutritional status can be maintained in good condition, delaying the progression of the disease, and preventing complications and sequelae. It is possible to increase resistance to the onset of the disease and prolong life.

Furthermore, administration of the composition of the present invention has the effect of suppressing body protein decomposition and promoting protein synthesis during biological invasion, and maintains a good nutritional state, so it can also be used as a nutritional supplement during biological invasion. suitable.

EXAMPLES Hereinafter, the present invention will be explained in more detail by showing reference examples, working examples, and test examples.

Reference Example 1 Potassium hydroxide was added to corn protein suspension (gluten liquor) 4501 obtained from the wet milling process to adjust the pH to 5.5, and a raw starch degrading enzyme was used as "Daviase" (trade name, manufactured by Daikin Industries) )70
g was added thereto, and the mixture was reacted at 50° C. for 16 hours with stirring.

The suspension was separated into solid and liquid using a filter press, the water-soluble fraction was removed, and a corn protein wet cake 8 was obtained.
Obtained 0 kg. This wet cake was resuspended in 350 g of distilled water, and the temperature was raised to 80° C. while stirring. Next, potassium hydroxide was added to adjust the pH to 12.0;
It was heat-treated at ℃ for 5 minutes.

Next, the suspension was cooled to 50°C, and the alkaliphilic bacteria (
80 g of high alkaline protease derived from Bacillus (No. 221) (manufactured by Meito Sangyo Building) was added and reacted for 1.5 hours. Furthermore, hydrochloric acid was added to the suspension to adjust the pH to 7.
0 and neutral protease “Bunazyme AP”.
180 g of J (trade name, manufactured by Nagase Biochemical ■) was added and reacted at 50°C for 200 hours. The reaction solution was separated into solid and liquid by filter press, and the water-soluble fraction was concentrated and Bx2
0.160 fl of corn protein enzymatic hydrolyzate was obtained.

A part of the above hydrolyzate was adjusted to pH 4.0 with hydrochloric acid, and then a polystyrene adsorption resin [DOWEX S-112J] was added.
(Product name: Dow Chemical Co., Ltd.) was activated with hydrochloric acid, and the hydrolyzate was passed through this column to separate a fraction with low aromatic amino acids. After concentrating this permeate, it was desalinated using an ion exchange membrane (Micro Acylyzer, manufactured by Asahi Kasei), treated with activated carbon, and freeze-dried to a total nitrogen concentration of 14.30%.
, amino nitrogen 3.00%, sugar content 5.20%, ash content 1.
A white powder was obtained having a composition of 0% and a molar ratio of branched chain amino acids to aromatic amino acids of 38.3. The amino acid composition is shown in Table 1.

Reference Example 2 The neutral protease in Reference Example 1 [Bunazyme A
Same as Reference Example 1, except that PJ (trade name, manufactured by Nagase Biochemical ■) was replaced with the acidic protease "Prothiase Amano MJ (trade name, manufactured by Ohno Pharmaceutical ■), and the reaction pH at that time was changed to 4. Repeat this operation, freeze-dry,
Total nitrogen 13.50%, amino nitrogen 2.95%, sugar content 5
．． A white powder was obtained having a composition of 0.00%, ash content of 1.2%, and a molar ratio of branched chain amino acids to aromatic amino acids of 69.1. The amino acid composition is shown in Table 1.

Reference Example 3 The same operation as in Reference Example 2 was carried out, the fraction with low aromatic amino acids was separated by passing through the adsorption resin, the pH was adjusted to 5.5 with potassium hydroxide, and the solid content concentration was 15% ( B x 1
5). After cooling the concentrate at 5°C, the precipitated tyrosine was removed by filtration, and then desalted, treated with activated carbon, and freeze-dried in the same manner as in Reference Example 2 to obtain a total nitrogen concentration of 13.8%.
Amino nitrogen 2.50%, sugar 4.5%, ash 1.3
%, and the molar ratio of branched chain amino acids to aromatic amino acids was 28.5.

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

Reference Example 5 Perform the same operation as in Reference Example 3, pass the tyrosine-removed fraction through a cation exchange resin and an anion exchange resin, separate the eluted fraction, and adjust the pH to 4.0 with potassium hydroxide. After that, it was desalted, treated with activated carbon, and lyophilized in the same manner as in Reference Example 1 to obtain a total nitrogen concentration of 14.0% and an amino nitrogen concentration of 0.90%.
%, sugar content 5.0%, ash content 1.0%, and the molar ratio of branched chain amino acids to aromatic amino acids is 25.6.
A white powder was obtained. The amino acid composition is shown in Table 1.

Reference Example 6 Perform the same operation as in Reference Example 1 up to the enzymatic reaction with the high alkaline prosthesis, and add hydrochloric acid to the reaction solution to adjust the pH.
Adjusted to 5.5. The reaction solution was applied to a filter press to separate solid and liquid, and the water-soluble fraction was concentrated to Bx15.
0 g of corn protein enzymatic hydrolyzate was obtained.

Next, in the same manner as in Reference Example 1, the section that passed through the adsorption resin was passed through a cation exchange resin and an anion exchange resin, the eluted section was fractionated, the pH was adjusted to 5.5 with potassium hydroxide, and the Salt, activated charcoal treatment, and lyophilization, the composition is 14.1% total nitrogen, 0.55% amino nitrogen, 4.8% sugar, and 0.9% ash, and has a composition of 14.1% total nitrogen, 0.55% amino nitrogen, 4.8% sugar, and 0.9% ash. A white powder with a molar ratio of 21.1 was obtained. The amino acid composition is shown in Table 1.

Note that Table 1 does not include data on tryptophan, which is a type of aromatic amino acid, because almost no tryptophan was included.

From the results in unpublished Table 1, the enzyme hydrolysates obtained in Reference Examples 1 to 6 have a ratio of branched chain amino acids (leucine, isoleucine, valine) to aromatic amino acids (phenylalanine, tyrosine) in the amino acid composition. It can be seen that is significantly higher. Further, it can be seen that in Reference Examples 3 to 6, branched chain amino acids were contained in a proportion equal to or greater than that of the starting material. In addition, it has a high protein content with a total nitrogen content of 14, and an amino nitrogen content of 3.0 or less, indicating that it 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) to determine the molecular weight distribution of each of the peptide compositions.

rOHPak KB-802゜5 as a separation column
(trade name, manufactured by Showa Denko ■) using an eluent of 0-1% trifluoroacetic acid-40% acetonitrile at a flow rate of 0. 5 mfl/min, and the detector is UV210n.
The analysis was conducted as m. In addition, as standard samples, thyrochrome C (MW Misaki 10.000) and oligopeptide G ly -G Iy -G Iy (MW 418
9.2), Gly-Gly (MW6132), and free amino acids tyrosine (MW4181°2) and phenylalanine (MW; 165.2) were used.

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 Figures 1 and 2, complete separation and identification of free amino acids and peptides has not been achieved, but considering that the minimum molecular weight of peptides is usually around 200, Reference Example 1
It can be seen that the molecular weight of the peptide contained in the peptide compositions of 3 and 3 is in the range of 200 to 4000.

Example 1 Using the peptide-containing substance obtained in the above reference example, a nutritional supplement composition of the present invention was prepared by mixing each component so as to have 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 effects of improving amino acid patterns, Fisher ratios, etc. by the peptide-containing substances contained in the compositions of the present invention.

Test Example-I A peptide-containing 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 other component amounts are as shown in Table 3.

6-week-old SD male and female rats BCAA peptide I administration group (
28 animals: weight 216.4±9.1g), and BCAA peptide ■ administration group (19 animals weight 218.4±9.8g)
The experiment was conducted in two groups. After fasting the rats for 18 hours, the test substance was orally administered. The test substance has a protein content of 1
．． The dose was 35 g/distillate, dissolved in 1011 LQ of ion-exchanged water, and administered using a catheter. Before administration, 30 days after administration
minutes, 60 minutes, 120 minutes, 180 minutes, 240 minutes, and 3
At 60 minutes, blood was collected from the aorta under ether anesthesia.

Blood was immediately centrifuged and plasma was stored at -80°C.

For the sample for amino acid analysis, 200 μg of plasma was mixed with an equal amount of 6% sulfosalicylic acid, centrifuged at 3000 rpm for 10 minutes, the supernatant was frozen overnight, thawed, centrifuged again, and the supernatant was used. As a coloring reagent, Ninhydrin reagent L-8500 set manufactured by Wako Pure Chemical Industries, Ltd. was used. The eluent is MCI BUFFERL manufactured by Mitsubishi Chemical Corporation.
-8500-PF kit was used. The amino acid analyzer used was a new model L-8500 high-speed amino acid analyzer manufactured by Hitachi. D, X60+a layer column: This column was packed with Hitachi custom ion exchange resin (#2622 SC). The results regarding blood amino acids are shown in FIG.

In addition, in the table, amino acids are IUPAC.

The IUB regulations or symbols commonly used in the field shall be followed, examples of which are listed below. Furthermore, when an amino acid or the like can have optical isomers, the L-isomer is indicated unless otherwise specified.

Leu: leucine, He: isoleucine, Ala: alanine, Glu: glutamine, Thr: threonine, H
is: histidine, Ser: serine, Gly nigericin, Asn: asparagine, Arg: arginine, Asp
: Aspartic acid, Pro Nibroline.

Part 3 Table Test Example ■ Portocaval anastomosis surgery (pc
Chronic liver failure model rats (group A, 2
5) and a 6-week-old SD male that underwent sham surgery (U)
(Group B, 25 mice) were kept for 2 weeks.

For these rats, changes in plasma free amino acid concentration and brain free amino acid concentration after a single administration of a peptide-containing substance were examined using the following method.

The peptide-containing substances administered are as shown in Table 4 below.

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

Based on these results, we evaluated the peptide-containing substances derived from corn gluten and found that the total amino acid concentration in blood,
It has been found that the above-mentioned peptide-containing substance has a high BCAA concentration and E/N ratio, and has a high nutritional supplementation effect and can improve the amino acid pattern in the blood, Fisher's ratio, etc. It can be seen that the composition of the present invention, which contains compounds, carbohydrates, and lipids in a well-balanced manner, has a high nutritional support effect and is particularly useful for nutritional support for liver disease patients, muscular dystrophy patients, and during biological invasion.

No. table

[Brief explanation of drawings]

Figure 1 is a chart showing the elution curve when the peptide composition obtained in Reference Example 1 was subjected to high performance liquid chromatography, and Figure 2 is a chart showing the elution curve when the peptide composition obtained in Reference Example 3 was subjected to high performance liquid chromatography. FIG. 3 is a graph showing the temporal change in blood amino acid concentration in Test Example I, and FIG. 4 is a graph showing the elution curve of B in Test Example ■.
FIG. 5 is a graph showing the temporal change in the CAA concentration. FIG. 5 is a graph showing the temporal change in the BCAA/AAA ratio in Test Example (2). (That's all) Figure 4 Sabakai (10,000) Figure 8 Reimon (Rei) 5 0 20 days 1 period

Claims (8)

[Claims] (1) A peptide-containing substance containing a peptide with a molecular weight of 200 to 4,000 obtained by hydrolyzing corn protein with an enzyme and having an amino acid composition of 3.6 mol% or less of aromatic amino acids based on the total amino acids, by dry weight. A composition containing 5 to 20% by weight on a dry weight basis, wherein the content of total nitrogen compounds, carbohydrates and lipids as a protein source in the composition is 15 to 4% by weight on a dry weight basis.
0% by weight, carbohydrates 50-70% by weight and fats 0-15
% by weight. (2) Claim (1) which is a nutritional supplement composition for liver disease patients.
The composition described in . (3) The composition according to claim (1), which is a nutritional supplement composition for patients with muscular dystrophy. (4) Claim (1) which is a nutritional supplement composition for use during biological invasion.
The composition described in . (5) The composition according to any one of claims (1) to (4), wherein the molar ratio of branched chain amino acids/aromatic amino acids in the amino acid composition of the peptide-containing substance is 10 or more. (6) In the amino acid composition of the peptide-containing substance, branched chain amino acids are 10 mol% or more and aromatic amino acids are 2 mol%.
The composition according to any one of claims (1) to (5), which is as follows. (7) The composition according to any one of claims (1) to (6), wherein the peptide-containing substance has the following amino acid composition. Amino acid composition (mol%) Aspartic acid 3.0-8.7 Threonine 2.1-6.2 Serine 3.4-9.6 Glutamic acid 10.9-30.4 Glycine 1.9-4.5 Alanine 8. 0-26.4 Valine 3.5-6.2 Cysteine 0.0-0.1 Methionine 0.4-6.5 Isoleucine 0.9-6.5 Leucine 4.4-47.5 Tyrosine 0.1- 2.0 Phenylalanine 0.0-1.6 Lysine 1.3-2.8 Histidine 1.0-2.5 Arginine 1.3-2.1 Proline 2.9-6.0 (8) The peptide-containing substance is 50 to 50% of the peptide in the solid content.
92% by weight, carbohydrates 5-20% by weight, amino acids 0-2
The composition according to any one of claims (1) to (7), which contains 5% by weight and 3 to 5% by weight of others.