JPS5976022A - Agent for reducing cholesterol value in blood - Google Patents

Abstract

PURPOSE:An agent for reducing the cholesterol value in blood containing a low- molecular peptide composition consisting of a dipeptide and tripeptide as main constituent components. CONSTITUTION:An agent for reducing the cholesterol value in blood containing a low-molecular peptide composition, having <=700 average molecular weight, preferably reduced to <=500, containing <=20wt% peptide content having >=700 molecular weight and <=20wt% free amino acid content, and consisting of a dipeptide and tripeptide as main constituent components. The resultant composition is capable of increasing the total nitrogen absorption rate with a small absorption antagonism of mutual amino acids, and the improved nitrogen balance provides a nitrogen source having a high efficiency. The body weight gain rate is remarkably increased. Any enzyme known at present alone is incapable of reducing the molecular weight, and various combinations, e.g. pepsin-''Molsin '' or pepsin-''Sanprose'', are effective for reducing the molecular weight to a desired value and the free amino acid content to a lower value.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a lowering agent for lowering blood cholesterol (am).

Conventionally, the production of peptides and amino acids by hydrolyzing proteins with enzymes has been carried out mainly in the food field. However, the purpose of these methods is to solubilize proteins by decomposing them with enzymes, or to reduce the molecular weight (molecular weight is several thousand or more) to make them suitable as food materials. There was no problem with the molecular weight itself.

The present inventors studied the relationship between enzymatic degradation products of proteins and digestion and absorption.

(1) lowering the average molecular weight to 700 or less, preferably 500 or less; (2) lowering the molecular weight to 700L/).

The content of the above peptide should be 20% by weight or less, (
3) reducing the content of free amino acids to 20% by weight or less;
That is, it was confirmed that a low-molecular-weight peptide composition containing dipeptides and tripeptides as main components lowers blood cholesterol levels. Furthermore, this composition.

(1) The intestinal absorption capacity is different from that of proteins or amino acid mixtures with the same amino acid composition, the total nitrogen absorption rate increases, and the absorption antagonism between amino acids is small.

(2) Improved nitrogen balance and becomes a highly efficient nitrogen source.

(3) The rate of weight gain increases significantly.

Many advantages such as those mentioned above were also confirmed. A diet with the same amino acid composition as shown in Table I below was prepared, which was amenable to the tests demonstrated in detail below.

Table I The nitrogen source in the above table has the following composition.

A...I Egg white protein B...■ Average molecular weight 420. Peptide composition C containing W free amino acids 8% by weight Free amino acid mixture D...■ Average molecular weight 1400. A peptide composition containing 2% by weight of m-isolated amino acids Each of these diets was given ad libitum to 10 Wistar rats for 2 weeks, and the results are shown in Table H.

Table ■ In the table above.

Food efficiency means that each
light Ga1n/ Food in take
It displays the ratio based on the value ■. The following can be understood from this table. The blood cholesterol level lowering agent (II) of the present invention has a significantly larger nitrogen retention amount (balance) than other drugs, and as a result, has a high food efficiency.
It was found that the blood cholesterol level was lowered even though the blood cholesterol level was significantly increased and the body weight was increased by 1%.

Furthermore, in order to consider the absorption of each amino acid rather than the macroscopic consideration of the nitrogen source, the nitrogen source sample (T) used in the diet described above was forcefully inserted into the stomach of Wistar rats that had been fasted for 24 hours (T). I
I), (II[) and (IV) were injected, blood was collected from the portal vein at regular intervals, the amino acid concentration was measured, and the amount absorbed per hour was measured. The results are shown in Table■ and Table■
Shown below. Note that the results are the average values of five rats each. Table ■ shows the average absorption rate of each amino acid until the absorption amount reaches its peak value, and Table ■ shows a comparison of the amino acid absorption patterns between sample (X) with almost ideal amino acid composition and samples (1) to (IV) above. It is a table.

Table ■ As is clear from this table, the blood cholesterol level lowering agent (II) of the present invention is not absorbed incompletely like egg white protein (I), and the absorption of amino acids is higher than that of the amino acid mixture (Iff). The degree of absorption antagonism is not large, and the initial absorption rate is about 30% higher than that of conventional protein decomposition products (IV).

Table ■ It is desirable that the absorption of amino acids be close to the ideal amino acid pattern (X). However, Table 1 shows that in egg white protein N) and amino acid mixture CIII), especially Phe T
It is clearly shown that the perpendicular rate of yr and old S from the ideal absorption pattern (X) is large. It was confirmed that the blood cholesterol level lowering agent (H) of the present invention has a close to ideal absorption pattern and achieves balanced absorption. The above facts are presumed to have a great influence on the metabolism of the absorbed amino acid itself or other amino acids.

One of the evidences for this seems to be a decrease in cholesterol levels.

As can be seen from the above test results, attention has not been focused on low-molecular-weight peptides that clearly lower blood cholesterol levels. Therefore, an object of the present invention is to provide a low molecular weight peptide composition that lowers blood cholesterol levels.

The present invention is an average molecule mainly composed of dipeptides and tripeptides with a free amino acid content and a peptide content of 700 or more molecular weight of 20% or less! This is a blood cholesterol level lowering agent whose main ingredient is a low molecular weight peptide composition of 700 or less.

Furthermore, preferably free amino acid content and molecules! 7
This is a blood cholesterol level lowering agent in which the content of peptides of 00 or more is 10% or less.

Furthermore, preferably, it is a blood cholesterol level lowering agent having an average molecular weight of 500 or less.

Hitherto, no problems have been raised regarding low-molecular-weight peptides mainly composed of dipeptides and tripeptides, which have many advantages as concluded from the test results listed in Tables I to (2) above. The present inventors attempted to produce such low-molecular-weight peptides from protein raw materials of arbitrary origin, and as a result, the following became clear (9)
(See table ■).

(1) Among protein hydrolases, pepsin has the strongest solubility, but degradation with pepsin does not proceed easily once the molecular weight reaches a certain level, and it is extremely difficult to reduce the average molecular weight to less than 1000. Have difficulty.

(2) The degrading power of neutral protease is weaker than that of acidic protease, and there is no enzyme other than pronase that can reduce the average molecular weight of the product to 1000 or less. However, because it is a complex enzyme, the production of free amino acids is extremely large, and at the stage when the average molecular weight approaches 500,
0% or more is free amino acids.

(3) Acidic protease is superior in terms of decomposition power2
Morsin (Fujisawa Pharmaceutical Origin Aspergi I Ius
saitoi) = Sanprose F (Adult Kyoei Bussan
Origin: Rh1zopus chinensis) etc. are useful as they produce less free amino acids.

(4) Although none of the currently known enzymes alone can reduce the molecular weight sufficiently to the desired extent, and the combination of Ceese's enzymes can sufficiently increase the molecular weight to the desired extent, these examples illustrate the present invention. However, the present invention is not limited to these, and various changes can be made thereto.

0 [Production Example I] 50 g of dried egg white (protein content 82 wt%) 11
of water, adjust the pH to 3 with hydrochloric acid, and dissolve 1 molsin in 1 ml of water.
g and 1 g of Sunprose F were added, and the reaction was carried out at 40° C. for 24 hours while maintaining Pll at 3. 10% of the reaction solution
After heating at 0°C for 10 minutes to inactivate the enzyme,
The mixture was centrifuged at 0 r, p, m (1500 G) for 10 minutes to remove insoluble matter, and the supernatant was freeze-dried. The yield of this product is 93.2% based on the raw protein,
The average molecular weight was 340. From the results of gel filtration of this product, a blood cholesterol level lowering agent of which 88-t% or more had a molecular weight of 700 or less could be produced. In addition, 700
The following distinctions cannot be made by gel filtration: The free amino acid content in the product was 8.5%.

[Production Example ■] Under the same conditions as Production Example ■ except for the enzyme used, first 1 g of morsin was added, then after 8 hours, 1 g of Sunprose F was added and the mixture was reacted for a further 10 hours, followed by the same treatment as in Production Example I. Did. The yield of the obtained product was 93.9%, the average molecular weight was 350, the content of M-free amino acids was 8.1%, and the gel filtration results showed that more than 90% had a molecular weight of 700 or less.

[Manufacturing example■]

Pepsin Ig was prepared under the same conditions as in Production Example I except for the enzyme used.
and 1 g of morsin were added at the same time, and the same treatment as in Production Example I was carried out. The yield of the product obtained was 96.1%, and the average molecular weight was 510. The free amino acid content was 7.8%, and the gel filtration results showed that more than 86% had a molecular weight of 700 or less.

[Manufacturing example■]

The same conditions as in Production Example I were used except for the enzymes used. First, 1 g of pepsin was added, and after 6 hours, 1 g of 1 morsine was added. The reaction was continued for another 10 hours, and then the same treatment as in Production Example I was carried out.

The yield of the product obtained was 98.3%, and the average molecular weight was 55.
0. The free amino acid content was 7.3%, and gel filtration results showed that more than 83% had a molecular weight of 700 or less.

[Production example V]

The same treatment as in Example I was carried out under the same conditions as in Production Example (1) except for the enzyme used, with 1 g of Vepsin and 1 g of Sunprose Fe-1 added at the same time. The yield of the obtained product was 94%, the average molecular weight was 430.3 and the amino acid content was 8.7%, and the gel filtration results showed that more than 90% had a molecular weight of 700 or less.

(Production Example ■) Under the same conditions as Production Example ■ except for the enzyme used, first 1 g of pepsin was added, then after 6 hours, 1 g of Sunprose F was added and the mixture was reacted for another 10 hours, followed by the same treatment as in Production Example I. The yield of the obtained product was 96%, the average molecular weight was 410. The free amino acid content was 9.2%, and the gel filtration results showed that more than 91% had a molecular weight of 700 or less.

[Manufacturing example■]

The conditions were the same as in Example I except for the enzyme used. First, 0.25 g of pepsin was added, and 3 hours later, 1 g of morsin was added.

After another 3 hours, add 1g of Sunprose F and add 1g of Sunprose F.
After reacting for a period of time, the same treatment as in Example I was carried out. The yield of the product obtained was 95%, and the average molecular weight was 370. The free amino acid content was 11.3%, and the gel 3 filtration results showed that more than 93% had a molecular weight of 700 or less.

The evaluation method of the products in the above examples is as follows.

(1) Product yield Amount of nitrogen in the product 100 Amount of nitrogen in raw materials Nitrogen was analyzed by Kjeldahl analysis method.

(2) Average molecular weight of product [average molecular weight of amino acids in raw protein] [product 1
[Number of moles of amino groups in 1 g of product] ×□ [Number of moles of amino groups in 1 g of product]
By the zen-3 ulfonic acid) method,
Complete hydrolysis of the product was performed in 6N HCl at 110
℃ for 24 hours by hydrolysis.

(3) Free amino acid quantification The product solution was treated with basic copper carbonate to form a copper complex with amino acid 4 and the peptide, which was adsorbed onto an anion exchange resin, and the free amino acid was eluted with 0.05M borate buffer. was quantified using an automatic amino acid analyzer. However, since acidic amino acids were not liberated by boric acid buffer, the product was directly subjected to an amino acid analyzer for quantification.

Accurate quantification is possible because there is no influence of peptides at the separation position of acidic amino acids in an amino acid analyzer.

(4) Determine the ratio of peptides with a molecular weight of 700 or less using 5ephadex G-10, which has the lowest molecular weight fraction of gel filtration.

In the manufacturing example, the raw protein is egg white, but is not limited to this and can also be ossein. In addition to soybean, wheat gluten, fish meal, chlorella, yeast protein, etc., protein substances enriched with specific amino acids by plastin reaction can also be used. The substrate concentration of the raw protein is 5-2
It is preferable to set it to about 0 w/v%. This is 5%
If it is less than 20%, it is not practical, and if it is more than 20%, it becomes too viscous5. The amount of enzyme to be added is 1 wt% or more, preferably 2 to 5 wt%, based on the substrate to achieve a degree of decomposition suitable for the purpose.
wt% is good. Reaction time is substrate concentration.

It is a function of the amount of enzyme, the degree of reactivity, etc., and the time is set at a time at which a peptide composition that does not degrade into amino acids can be obtained.

The reaction temperature is determined depending on the optimum temperature of the enzyme used. The acid used may be a strong acid or a weak acid. Table 3 below shows a case where a protein is degraded by a combination of two or more proteases as in the production example of the present invention and a case where a single protease is used for degradation.

Table 7 6 Comparison of Examples and Table V above 1 According to the method of the present invention, the target dipeptide and tripeptide with a molecular weight of 500 as the main component can be obtained from various protein sources in high yield.
It is broken down into the following, and the content of free amino acids is 10w
It is easy to understand that low-molecular-weight peptides with characteristics such as low t% or less, low amino acid absorption antagonism, and relatively high-molecular peptide content with a molecular weight of 700 or more and a low content of 20% or less can be reliably produced. can.

Patent Applicant: Terumo Co., Ltd. 1-day Procedural Amendment Application: Ro/V/1980: Mr. Kazuinu Wakasugi, Commissioner of the Patent Office], Case Description: 1980 Patent Application No. 110741 2, Name of the Invention: Lowering Blood Cholesterol Levels Agent 3, Person making the amendment 4, Date of amendment order Voluntary (1) Amend the name of the invention in the specification to "blood cholesterol level lowering agent."

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Claims (2)

[Claims] (1) Blood cholesterol level reduction using a low-molecular-weight peptide composition with an average molecular weight of 700 or less, which contains dipeptides and tripeptides as main components and has a free amino acid content and a peptide content of 700 or more in 20% or less agent (2) The blood cholesterol level lowering agent according to claim 1, wherein the content of free amino acids and the content of peptides having a molecular weight of 700 or more are 10% or less. (3) The average molecular weight is 5.
The blood cholesterol level lowering agent according to claim 1 or 2, which is 00 or less.