WO2004103391A1

WO2004103391A1 - Hepatic lipid metabolism accelerating agent, hepatic lipid metabolism accelerating health food and process for producing peptide mixture as active ingredient thereof

Info

Publication number: WO2004103391A1
Application number: PCT/JP2004/006525
Authority: WO
Inventors: Nobuhiro Fukuta; Masanobu Sakono; Nobuko Iritani; Toshihiro Nakamori; Hitoshi Furuta; Michihiro Sugano
Original assignee: Fuji Oil Company, Limited
Priority date: 2003-05-22
Filing date: 2004-05-14
Publication date: 2004-12-02
Also published as: JPWO2004103391A1

Abstract

A hepatic lipid metabolism accelerating agent or hepatic lipid metabolism accelerating health food that is effective in acceleration of hepatic lipid metabolism and thus amelioration to or prevention of, for example, life-style related diseases associated with hepatic lipid metabolism abnormality. In particular, a hepatic lipid metabolism accelerating agent or hepatic lipid metabolism accelerating health food, comprising as an effective ingredient a mixture of hydrophilic peptide wherein 30 to 70 wt.% of the constituent amino acids consist of acidic amino acids. There is further provided a process for producing a peptide mixture, characterized by decomposing a protein with the use of at least one type of protease and treating the decomposition product by means of a hydrophobic resin.

Description

Specification

Method for producing liver lipid metabolism promoter, liver lipid metabolism promoting health food, and peptide mixture as active ingredient thereof

Technical field

The present invention provides a liver lipid metabolism promoting agent, a liver lipid metabolism promoting health food and a peptide mixture as an active ingredient thereof.

Background art

[0002] The liver is the largest among human organs and corresponds to about 1/50 of body weight. Its physiological functions are extremely diverse and play a central role in carbohydrate, protein metabolism, and lipid / energy metabolism, and are indispensable for maintaining the life of living organisms, such as storage of substances, detoxification, hematopoiesis, and blood coagulation. There are many.

[0003] The lipid metabolism function assigned to the liver cells is deeply linked to the lipid metabolism function in the body, and one of the causes is a decrease in the lipid metabolism function of the liver and further dysfunction, resulting in hyperlipidemia, hypertension, and dysfunction. Diseases such as pulse sclerosis, obesity, fatty liver, diabetes and hypertension are caused. To improve the dysfunction of lipid metabolism in the liver, it is considered important to promote lipid metabolism and reduce the concentration of lipid accumulated in cells.

[0004] In recent years, attention has been paid to the preventive and therapeutic effects of ingredients contained in foods, and the preventive and therapeutic effects of food ingredients can be obtained for the aforementioned circulatory diseases. Development is expected.

[0005] Lipid metabolism improvers derived from some proteins and their hydrolysates (peptides) are already known. It is known that vegetable proteins, particularly soy protein, are superior to animal proteins in terms of the effect of proteins on improving lipid metabolism. As for these protein hydrolysates (peptides), the following are known, including those derived from proteins and proteins, and their functions and effects are various.

[0006] First, as a lipid improving agent other than the hydrolyzate of soybean protein, for example,

JP-A-52-83816 (new polypeptide), JP-A-04-149137 (diet), JP-A-06-211690 (feeding for casein-derived blood lipid suppression), JP-A-11-80006 JP (Lipid Absorption Inhibitor), JP-A-2000-264845 (Fish-Derived Cholesterol-Lowering Agent), JP-A-2000-228967 (Lipid Metabolism Processed Food), JP-A 2001-2577 (Derived from Royal Jelly) JP-A-2001-57868 (material for improving fat metabolism derived from wastewater of livestock processing), JP-A-2001-57869 (material for improving obesity and diet), JP-A-2001-58955 JP-A-08-157389 (a therapeutic agent for hypertriglyceridemia), JP-A-09-157290 (cholesterol-reducing peptide), JP-A-09-255698 ( Peptides inhibiting blood triglyceride concentration increase) and JP-A-07-188284 (peptides inhibiting blood triglyceride concentration increase) are known.

[0007] On the other hand, it is known that the effect of improving lipid metabolism is also various for peptides obtained by hydrolyzing proteins. For example, Japanese Patent Application Laid-Open No. 5-87052 states that a lipid metabolism improving agent has an effect of lowering the fat content and the amount of lipid droplets in the liver. This is due to a decrease in the fat content absorbed and a decrease in the serum fat content, which is not an effect of altering the lipid metabolic activity itself in the liver.

Japanese Patent Application Laid-Open No. 04-149137 (diet) is known as a proteolyzed low molecular weight peptide, which is composed mainly of dipeptide and tripeptide, and thus has an inhibitory effect on fat accumulation and weight reduction. It is said that it has an effect on suppressing the increase in lipid metabolism in the liver.

As a physiologically active composition, Japanese Patent Application Laid-Open No. 10-203994 discloses that a peptide having a molecular weight of 500-5000 derived from a soybean protein hydrolyzate has an effect of increasing neutral fat in blood and HDL-cholesterol. As mentioned, it promotes fatty acid metabolism in the liver.

[0008] In recent years, soybean protein is a food material which has excellent nutritional properties among vegetable proteins, has various physiological activities recently, and is attracting attention as a physiologically functional agent. Although it is known that soybean protein has an effect of suppressing obesity, the mechanism of lowering the body fat percentage is known to be due to the suppression of the activity of fatty acid biosynthetic enzymes in the liver (Iriya J. Nutr., 126, 380, 1996), and it is not known which component in soy protein is the active ingredient. [0009] The present applicant has continuously studied the physiological action of soybean protein hydrolyzate (peptide mixture), particularly on lipid metabolism. For example, as disclosed in Japanese Patent Publication No. 7-025796, soybean protein hydrolyzate (oligopeptide mixture) suppresses a rise in blood cholesterol, and is disclosed in Japanese Patent Application Laid-Open Nos. 1-269456 and 3-272694. Has been disclosed to be involved in cholesterol and triglyceride metabolism as disclosed in US Pat. However, among lipid metabolism in the body, it was not clear which peptide promotes lipid metabolism in the liver and improves lipid concentration in the liver. In addition, the fact that the triglyceride content in the liver of a soybean protein hydrolyzate is reduced is described only in the cited data in JP-A-4-51872. It is presumed that soybean protein and its hydrolyzate contain components that suppress the activity of liver fatty acid biosynthetic enzymes, but it has not yet been determined which components exert their effects. Power, ivy.

[0010] In recent years, attempts have been made to further purify hydrolyzed peptides and fractionate functional components. For example, Japanese Patent Application Laid-Open No. 07-188284 discloses that a fraction having a high hydrophobic amino acid content, fractionated by ion exchange resin + ultrafiltration, and further reversed phase chromatography, has a blood TG rise inhibitory effect. . JP-A-2002-212097 discloses a lipid metabolism-improving agent for a milk-derived basic peptide fraction, which is produced using a cation exchanger, and has a glutamic acid + aspartic acid content of 30%. The following is different from the present invention. There is no known lipid metabolism promoter in the liver that does not feel bitter and has a high acidic amino acid content as in the present invention.

Disclosure of the invention

Problems the invention is trying to solve

[0011] The present invention promotes lipid metabolism in the liver, and improves or prevents lifestyle-related diseases associated with abnormal lipid metabolism in the liver. Aimed to provide.

Means for solving the problem

[0012] The present inventors have conducted intensive studies to solve the above-mentioned problems, and obtained a finding that a peptide mixture rich in acidic amino acids (glutamic acid-aspartic acid) and having a large ratio promotes lipid metabolism in the liver. Thus, the present invention has been completed. That is, the present invention is a lipid metabolism-promoting agent in liver comprising a hydrophilic peptide and a peptide mixture in which 30 to 70% by weight of amino acids constituting the hydrophilic peptide are acidic amino acids as an active ingredient. Proline in the constituent amino acids of the hydrophilic peptide is preferably 5% or less of all the amino acids. More specifically, it is an enzymatically decomposed product of a protein, having an average molecular weight of 200-10 000, an acidic amino acid of 30 70% by weight (hereinafter referred to as “%”), a basic amino acid of 10% -30%, and a branched amino acid in the total amino acid composition. A liver lipid metabolism promoter containing as an active ingredient a peptide mixture containing 15.0% or less of chain amino acids, 9.0% or less of aromatic amino acids, and 5.3% or less of proline is preferable. The present invention also provides a method for producing a peptide mixture, which comprises decomposing a protein with one or more proteolytic enzymes and treating the resultant with a hydrophobic resin. The protein is preferably soy protein. As the hydrophobic resin, a styrenedibutylbenzene resin is preferable.

The present invention is also a health food which promotes lipid metabolism in the liver, comprising a peptide mixture comprising a hydrophilic peptide, wherein 30 to 70% by weight of the constituent amino acids of the hydrophilic peptide is an acidic amino acid as an active ingredient. Proline in the constituent amino acids of the hydrophilic peptide is preferably 5% or less of the total amino acids. More specifically, it is an enzymatically decomposed product of a protein, having an average molecular weight of 200-10000, an acidic amino acid content of 30-70% by weight (hereinafter referred to as "%"), a basic amino acid content of 10% -30%, and a branched amino acid. Healthy foods that promote lipid metabolism in the liver and contain a peptide mixture containing 15.0% or less of chain amino acids, 9.0% or less of aromatic amino acids, and 5.3% or less of proline as active ingredients are preferred.

The invention's effect

[0013] The present invention provides an oligopeptide mixture that promotes hepatic lipid metabolism and inhibits hepatic fatty acid biosynthesis, resulting in abnormal hepatic triglyceride concentration, resulting in obesity, hypertriglyceridemia, and fat. Prevents lifestyle-related diseases such as liver, diabetes or hypertension, coronary artery disease, cerebral artery disease, chronic nephritis, nephrosis, cirrhosis, obstructive jaundice, etc. It is now possible to apply it to pharmaceuticals.

BEST MODE FOR CARRYING OUT THE INVENTION

First, the liver lipid metabolism promoter of the present invention will be described. Lipids in this liver The metabolism promoter is composed of a hydrophilic peptide, and contains, as an active ingredient, a peptide mixture in which 30 to 70% by weight of the constituent amino acids of the hydrophilic peptide are acidic amino acids. What is important here is that the lipid metabolism-promoting peptide mixture in the liver contains no water-insoluble fraction and is composed of hydrophilic amino acids, and that the proportion of acidic amino acids is high. As hydrophilic amino acids, glycinin (Gly), serine (Ser), threonine (Thr), cystine (Cys), asparagine (acid) (Asp), glutamine (acid) (Glu), lysine (Lys), S is the force that arginine (Arg) and histidine (His) are raised. The ratio of acidic amino acids in the amino acids constituting the hydrophilic peptide is 0% by weight (hereinafter simply referred to as%), preferably 35 55%. It is. Examples of the acidic amino acid include asparagine, aspartic acid, gnoretamine, and gnoretamic acid. Glutamine and glutamic acid are preferred. The higher the proportion of glutamic acid or glutamine, the better the effect of promoting lipid metabolism in the liver. If the proportion of gnoretamic acid or glutamine is less than 30%, the effect of promoting lipid metabolism in the liver, which is superior to that of the known soybean peptide mixture, is not exhibited.

Further, among the constituent amino acids, it is particularly desirable that the proline content be 5% or less, preferably 3% or less, particularly 1% or less from the viewpoint of inhibiting the synthesis of fatty acids. More specifically, it is an enzymatically decomposed product of a protein, having an average molecular weight of 200-10000, an acidic amino acid content of 30-70% by weight (hereinafter referred to as "%"), a basic amino acid content of 10% -30%, and a branched amino acid. A lipid metabolism promoter in the liver containing, as an active ingredient, a peptide mixture containing 10.5% or less of chain amino acids, 8.0% or less of aromatic amino acids, and 5% or less of proline is suitable. It is not clear whether the average molecular weight of this peptide mixture contributes to the effect of promoting lipid metabolism in the liver. However, the lower the gastrointestinal absorption rate, the faster it can be. Depending on the disassembly method, it can be 500 3000. Although the effect of the free amino acid content on the promotion of lipid metabolism in the liver is not clear, it is generally appropriate that a small amount of the peptide mixture is preferably 35% or less, preferably 10% or less. The effect of promoting lipid metabolism in the liver is not known, but in the present invention, the basic amino acids are 10% 30%, the branched-chain amino acids are 15.0% or less, and the aromatic amino acids are 9.0% or less. Desirably. In terms of the flavor and stability of the lipid metabolism promoter and health food in the liver of the present invention and the stability thereof, the smaller the content of hydrophobic amino acids, the more favorable the branched chain amino acids and aromatic amino acids. The above range is preferable.

[0016] The promotion of lipid metabolism in the liver of the present invention is to suppress the biosynthesis of fatty acids in the liver. That in the liver, a series of enzymes der synthesize fatty acids from Asechiru CoA Acety Bok and _{oA-carboxylase, Fatty acid synthase Λ} ATPcitrate-lyase, inhibit the enzymatic activity of such Malic enzyme ^ Glucose-6-phosphate dehydrogenase, and

Acety is a lipid metabolism that suppresses the expression of mRNAs such as CoA_carboxylase and Fatty acid synthase, and has the effect of reducing the triglyceride and cholesterol content accumulated in the liver.

[0017] Accordingly, the peptide mixture for improving lipid metabolism in liver of the present invention can be used as a pharmaceutical or health food for preventing or treating diseases based on the suppression of fatty acid synthesis. The lipid metabolism promoter in the liver used as a medicament can be contained as an oligopeptide mixture in an amount of 0.01% or more, preferably 0.1% or more. If it is less than 0.01%, it is difficult to realize a substantial effect. Further, the daily intake or dosage is appropriately 0.1 mg / kg / day or more, preferably 1 mg / kg / day or more, more preferably 10 mg / kg / day or more as an oligopeptide mixture. 0. If less than lmg / kg / day, the effect may not be observed. In addition, a health food that promotes lipid metabolism in the liver as a health food exerts the same effect at the same content and intake of the peptide mixture. In this manner, the liver lipid metabolism promoter and the liver lipid metabolism promoting health food of the present invention significantly promote liver lipid metabolism, and thus, have abnormal liver lipid metabolism, specifically, It can be used as an agent or food for the purpose of preventing or improving obesity, hypertriglyceridemia, fatty liver, diabetes or hypertension caused by abnormal triglyceride concentration. When used as food, it can be used as a solution or as a raw material for concentrated liquids, freeze-dried products and spray-dried products, depending on the form of oral nutritional food, tube nutritional food, etc. Foods include a wide variety of foods, such as beverages, frozen desserts, tablets, and confectionery. It can also be used as a health food as capsules or tablets that are not in the form of ordinary food.

Next, a method for producing a peptide mixture effective for promoting lipid metabolism in the liver of the present invention will be described. The mixture of peptides for promoting lipid metabolism in the liver according to the present invention converts soybean protein with an enzyme. The power that can be obtained by disassembling S As the soy protein used in the present invention, soy milk, concentrated soy protein, isolated soy protein, defatted soybean, soy whey protein, or the like can be used as an inexpensive material. Among them, isolated soy protein is preferable. The concentration of the soybean protein solution to be subjected to the enzyme treatment is suitably 1% by weight to 30% by weight, preferably 515% by weight, more preferably 812% by weight. A low concentration does not hinder enzymatic degradation, but is not preferred because it reduces productivity. If the concentration of the soybean protein solution is too high, the polymerization of the once hydrolyzed protein hydrolyzate will be strong, so a large amount of enzyme will be required to sufficiently decompose it, which is not preferable.

In the enzymatic decomposition, it is appropriate to hydrolyze soybean protein in an aqueous system (soybean protein slurry or solution) using an enzyme. The proteolytic enzyme (protease) used in the present invention may be an exoprotease or an endoprotease alone or in combination, and may be of animal, plant or microbial origin. Specifically, serine proteases (trypsin, chymotrypsin, animal-derived subtilisin, carboxypeptidase, etc.), thiol proteases (plant-derived papain, fusin, bromelain, etc.), carboxy proteases (animal-derived pepsin, etc.) ) Can be used. More specifically, “Protin FN” derived from Aspergillus oryzae (manufactured by Daiwa Kasei Co., Ltd.), Streptomyces “actinase” derived from Griseus (manufactured by Kaken Pharmaceutical Co., Ltd.), and “ Alcalase "(manufactured by Novozymes Japan Ltd.) and" Protin A "(manufactured by Daiwa Kasei Co., Ltd.) derived from Bacillus subtilis. In addition, examples of enzymes containing endoprotease include “proteases 3” (manufactured by Amano Pharmaceutical Co., Ltd.) and “Protin AC-10” (manufactured by Daiwa Kasei Co., Ltd.). An example of the proteolytic enzyme is “Proteases 1 ^” (manufactured by Amano Pharmaceutical Co., Ltd.).

[0020] The hydrolysis conditions of the present invention vary somewhat depending on the type of protease used, but generally, an amount sufficient to hydrolyze soybean protein in the action pH range and the action temperature range of the protease is used. Preferably, it is used. Considering that the intrahepatic lipid metabolism promoting peptide mixture of the present invention is used for a salt-restricted diet (eg, a tube feeding diet), if the pH is 510, preferably pH 6-9, the salt by neutralization is used. It is preferable because the generation of can be reduced.

[0021] Removing the insoluble decomposed product from the enzyme decomposed product of soybean protein facilitates the subsequent resin treatment. This is preferable. Means for separating and removing insoluble degradation products from the enzyme-decomposed solution of soybean protein may be by filtration means such as filter press or membrane separation, but most preferably a method of using both centrifugation and membrane separation. In the case of enzymatic digestion under acidic conditions, for example, when the pH of the enzyme digest of soybean protein is in the range of 3-8, to increase the separability during this separation, the pH must be adjusted to increase the cohesion of insoluble substances. 4-6.2, preferably 4.5-5. This is because insoluble matter including undegraded matter tends to aggregate around the isoelectric point of soybean protein. Alternatively, salts or hydroxides such as chlorides and sulfates of calcium magnesium in the decomposed solution, alkaline earth metal compounds, or protein coagulants such as sodium polyacrylate, alginic acid, chitin chitosan, etc. Separation can be enhanced even if squeezed.

[0022] As described above, after separating and removing insoluble decomposed products from the enzyme-decomposed solution of soybean protein, the hydrophobic component is adsorbed on a hydrophobic adsorbent such as polystyrene beads, whereby the hydrophilic component is recovered. Suitable hydrophobic adsorbing resin materials include, for example, Amberlite XAD (registered trademark) made of organonone earth, Levatit OC (registered trademark) manufactured by Bayer, and Diaion (registered trademark) manufactured by Mitsubishi Kasei. More specifically, as an aromatic resin, a styrene-vinylinolebenzene-based resin (for example, HP-20, HP-21, SP-825, SP-206, SP-207, SP-800 (all are registered trademarks, Acrylic resins (HP1MG, HP2MG (both are registered trademarks, manufactured by Mitsubishi Kasei Corporation), etc.), phenolic resins (S874, S861 (both are registered trademarks, Sumitomo Chemical ( Etc.) are suitable.

[0023] As an embodiment of treating with a resin to adsorb and remove the hydrophobic component, the resin can be brought into contact with a soybean protein enzyme digestion solution. This contact method does not matter even in a batch process or a continuous column process. For example, when the treatment is performed by the notch method, the amount of the hydrophobic adsorbent used is about 0.5 to 100 times the weight of the dry weight of the soybean protein hydrolyzate, and more preferably about 260 times the weight. If the amount of the adsorbent is small, the hydrophobic amino acid cannot be sufficiently adsorbed, and if the amount is too large, the yield of the other oligopeptide mixture decreases. After stirring in a temperature range of 140 ° C for about 5 minutes to 12 hours, the mixture can be filtered with a filter that can pass water-insoluble polymer components but cannot pass resin.

[0024] In the method using a continuous column, the soybean protein hydrolyzate is used in an amount of 1 part by weight of dry matter. 2 to 200 parts by weight, more preferably about 25 to 100 parts by weight, of the soybean protein hydrolyzate solution as a hydrophobic adsorbent, and a linear velocity at which the contact time with the hydrophobic adsorbent is 5 minutes or more. It can be. At this time, the concentration of the soybean protein hydrolyzate is preferably 2 to 50%, more preferably 10 to 30%. To regenerate the hydrophobic adsorbent, 0.53N sodium hydroxide or about 1090% of organic solvent can be used. Ethanol, methanol, isopropanol, acetone, etc. are used as the organic solvent. It can be used about 120 times the capacity of the material.

[0025] The soybean protein peptide mixture solution obtained as described above can be used as it is or after being concentrated depending on the intended use. However, usually, it is sterilized and then subjected to powder drying by freeze-drying or the like. It can be used in the state.

[0026] The peptide mixture obtained by the above method is hydrolyzed to analyze the amino acid composition. Compared with the original soybean protein hydrolyzate, the composition of the hydrophilic amino acid is higher and the composition of the hydrophobic amino acid is lower. . Among the hydrophilic amino acids, especially, the content of glutamic acid, which is an acidic amino acid, is high, and the content of aspartic acid tends to be high next. The content of basic amino acids and other hydrophilic amino acids does not change much. The hydrophobic amino acid content is reduced, but the proline is particularly reduced, and the higher the decrease, the lower the levels of phenylalanine, leucine, isoleucine and tyrosine. In addition, the peptide thus prepared is characterized by having a good flavor and little browning even after long-term storage.

Example

Hereinafter, embodiments of the present invention will be described with reference to examples.

(Example 1) Method for producing peptide mixture

3 kg of isolated soybean protein (Fuji Oil Co., Ltd., “New Fuji Pro-R”) is added to water to make a 10% aqueous solution, and proteolytic enzyme (Daiwa Kasei Co., Ltd., “Protin AC-10”) 120 g After hydrolyzing at 50 ° C for 5 hours (15% TCA solubility: 85%), the resulting precipitated components were separated and removed by a high-speed centrifuge. The obtained supernatant was sterilized at 140 ° C. for 7 seconds by blowing steam at a pressure of 8 kg m 2, and the solution was further filtered through a filter of 0.22 μm (manufactured by Cuno Inc.) and powder-dried by spray drying.

[0028] The peptide powder obtained above was prepared in a 10% aqueous solution, and the mixture was kept at 7 ° C for about 1 hour. The mixture was treated in a batch system with the biosynthetic adsorbent HP-20 (manufactured by Mitsubishi Kasei Co., Ltd.), and passed through a 1-micron filter paper to remove the resin, and the treatment liquid was freeze-dried. The weight of the dried peptide was 1, 2, 4, 8, 16 and 32 times the weight of the dried peptide, and the resin was treated. The average molecular weight of the peptide mixture obtained by the above method is 700 (gel filtration method using HPLC), the average molecular weight of the peptide after fractionation is 700 800, and the molecular weight tends to increase slightly as the resin ratio increases. It was in the direction.

The lyophilized product obtained in Example 1 was hydrolyzed with 6N hydrochloric acid at 110 ° C. for 24 hours, and then its amino acid composition was analyzed with an amino acid analyzer (L-8500, manufactured by Hitachi, Ltd.). The results are shown in Table 1.

^[ Table 1 ^] Hydrophobic α / acid with amino acid content%>

Minutes 鑕

Ala Val He Leu Pro Met Phe Tyr Tip

A A Start of agriculture 4.1 3.8 3.8 8 5.5 1.5 4.9 4.2 9.9 15.6 9.9

X 1 4 33 3.4 6.9 5.1 1J 4 3.5 0.5 13.6 8

X 2 4 3.5 3.6 6.8 5 1.2 3.6 3.4 0.4 13.9 7.4

X 4 4.2 3.4 3.2 6.4 4.5 1.2 3.3 3.1 03 13 6.7

X 8 4.7 3.4 2.2 53 2.4 1 2.1 .2 0.11 10.9 4.21

X 16 4.9 3.1 2 4.6 1.7 0.9 1.7 1.5 0.1 9.7 3J 32 4.9 3.1 1.7 3.9 0.9 0.8 0.8 1.1 0.05 8.7 1.45 128 6 0.9 0 0 0 0 0 0 0 0.9 0

Proficiency

Asp His acidic

Gly Ser Thr Cys Glu Lys

A 57 »a ^» start 3.8 5.6 4.1 1.1 12.1 19.5 6.2 8.1 3 31.6 173

X 1 3.6 5.5 4.1 1 12.6 24 6.2 8.1 2.8 36.6 17.1

X 2 3.7 5.5 4.1 1 12.7 24.7 6.2 8 2.7 37.4 16.9

X 4 3.6 5.6 4.2 1 12.7 26.7 6.5 7.6 2.6 39.4 16.7

X 8 3.4 6 4.2 0.7 13 33.8 6.4 6.8 2.7 46.8 15.9

X 16 3.2 6 4.1 0.6 13.4 36.2 6.7 6.7 2.6 49.6 16 32 3 6.1 4.2 0.6 13.8 39 6.7 6.7 2.6 52.8 16

X 128 0 9 4.5 0 18 42.1 7 9 3.5 60.1 19.5

(Example 2) Test on cultured cells Part 1 / Measurement of mRNA amount of fatty acid synthase

Rat hepatocytes obtained by the collagenase method were plated with WE medium (20 mM glucose, 10-7 M Insulin) for 5 hours, and then the samples were added. Change the medium in the morning and evening and after 24 hours Total RNA was extracted and the amount of mRNA was measured. WE medium is 2.04g start as amino acid, XI, X4, X8tt0.2g / L.

[0032]

(Table 2-1) mRNA levels of fatty acid synthase Acety CoA-Carboxylase

(Change amount when WE is set to 1)

WE 1.0

Start 0.94

X 1 0.39

X4 0.33

X 8 0.19

(Table 2-2) mRNA level of Fatty acid synthase

WE 1 0

Star 'ho 75

X 1 0 78

X 4 0 72

X 8 0 19

(Example 3) Test on cultured cells Part 2

Rat hepatocytes obtained by the collagenase method were plated with WE medium (20 mM glucose, 10-7 M Insulin) for 5 hours, and the samples were added. The medium was changed in the morning and evening, and harvested after adjusting for 48 hours. After homogenizing the hepatocytes, the enzymatic activity after 105,000 X g supernatant was measured. WE medium is 2.04 g / L as amino acid, start, XI, X4, X8 is 0.2 g / L ₀

(Table 3) Measurement of activity of fatty acid synthase

Acety ト CoA- carboxylase Fatty acid synthase ATPcitrate-lyase

WE 0 WE 0 WE 0

X 1 3 X 1 0.73 X 1 0.76 X 4 0 92 X 4 0.73 X 40,36 X 8 0 42 X 8 0.55 X 8 0,24

Glucose-6-phosphate

Malic enzyme dehydrogenase

WE 1.0 WE 1.0

X I 1.18 X I 0.76

X 4 0.76 X 4 0.24

X 80.67 X80.42

(Example 4) Test in animals (Test of lipid concentration in liver)

For the rats, male Sprague-Dawley (SD) rats (4 weeks old, weighing 70-80 g) were purchased from Kudo Co., Ltd. (Kumamoto). After 6-10 days of preliminary breeding, they are divided into four groups: casein group, soy protein group, soy protein enzyme digestion (starting material), and resin fraction of soy protein enzyme digest (X32 above) as shown below. Food for 4 weeks. The temperature of the breeding room was maintained at 22-24 ° C and the light period was from 7:00 to 19:00. Feed and deionized water were given ad libitum. After two weeks of feeding, rats were sacrificed by decapitation and ingested liver and adipose tissue. In addition, adipose tissue ingested fat around the kidney and epididymis. Rats weighing 130-140 g

Male rats of the Sprague-Dawley (SD) strain were allowed to freely take a diet prepared according to AIN76 for 2 weeks. The digestion of soy protein enzymatic products does not cause any problem in the feeding ability of rats, but the feed containing the fractionated product (X32 above) showed strong suppression of food intake, so 50% of the protein was reduced by casein. The test was performed in place of In the sample composition, the protein content is 20% casein, 10% casein + soybean protein ("Fujipro R" manufactured by Fuji Oil Co., Ltd.) 9.75%, casein 10% + soybean protein degradation product in terms of nitrogen 9. 12.26% of a resin fraction (X32 above) of 64% and 10% casein + enzyme hydrolyzate of soybean protein were added. In addition, corn starch 15%, DL methionine 0.3%, cellulose 5%, corn oil 5%, mineral mixture 3.5%, vitamin mixture 1%, choline bitartrate 0.2% are added, and sucrose is added to 100% Like Made

[0036]

(Table 4)

(Unit: fflg / g liver) Casein Soy protein Proteolysis start Proteolytic fraction

TG 84.5 Sat 6 a 16.2 +1.8 b 21.0 k 2.5b 18.2 ± 1.4 b PL 25.7 ± 0.7 a 28.6 ± 0.7 b 26.3 ± 0.3 a 26.4 ± 0.7 a TC 2.95 Sat 0.11a 1.89 k 0.07b 2.15 ± 0.09b 2.02 ± 0.08 b FC 1.55 person 0.09a 1.37 ± 0.03ab 1.32 ± 0.06b 1.33 ± 0.05b ester 47.1 person 3.8 a 26.7 ± 3.2 b 38.2 person 2.6 ac 34.0 ± 3.2 be Content (%)

(Mean soil standard deviation) abc Significant difference between different alphabets

(Example 5)

A lipid metabolism-improved beverage having the composition shown in Table 5 below was produced. The flavor of the produced beverage was good, and the flavor did not deteriorate even after storage at room temperature for one year, and there was no problem such as precipitation.

[0038]

(Table 5) 15% of freeze-dried product treated in 8 times the amount of Example 1

Reduced maltose syrup 20%

m 0%

Sucralose 0 0045% Fumaric acid 0 12%

Cuenoic acid 0 2%

water

Claims

The scope of the claims

[1] An agent for promoting lipid metabolism in the liver, comprising a peptide mixture consisting of a hydrophilic peptide, wherein 30 to 70% by weight of the constituent amino acids of the hydrophilic peptide is an acidic amino acid.

[2] The agent for promoting lipid metabolism in liver according to claim 1, wherein proline in the constituent amino acids of the hydrophilic peptide is 5% or less of all amino acids.

[3] An enzymatically degraded product of protein, with an average molecular weight of 200,000, 30-70% by weight (hereinafter%) of acidic amino acids, 10% -30% of basic amino acids, and branched-chain amino acids in the total amino acid composition. 3. The liver lipid metabolism promoter according to claim 1 or 2, comprising a peptide mixture containing not more than 15.0%, not more than 9.0% of aromatic amino acids and not more than 5.3% of proline as an active ingredient.

[4] A method for producing a peptide mixture, which comprises decomposing a protein with one or more proteases and treating with a hydrophobic resin.

[5] The production method according to claim 4, wherein the protein is soy protein.

6. The method according to claim 4, wherein the hydrophobic resin is a styrenedivinylbenzene-based resin.

[7] A health food which promotes lipid metabolism in the liver, comprising a peptide mixture comprising a hydrophilic peptide, wherein 30-70% by weight of the amino acids constituting the hydrophilic peptide are acidic amino acids.

[8] The health food for promoting lipid metabolism in the liver according to claim 1, wherein proline in the constituent amino acids of the hydrophilic peptide is 5% or less of all amino acids.

[9] Enzymatically degraded protein, with an average molecular weight of 200,000, 3070% by weight (hereinafter%) of acidic amino acids, 10% -30% of basic amino acids, and 1% of branched-chain amino acids in the total amino acid composition. 5.0. / o or less, 9.0 of aromatic amino acids. / o and below, 5.3 for proline. 9. The health food for promoting lipid metabolism in the liver according to claim 7 or 8, which comprises a peptide mixture having a ratio of / ₀ or less as an active ingredient.