JP2007228875A - Anti-fatigue peptide derived from meat protein - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an anti-fatigue agent or a functional food safe and effective for fatigue prevention or fatigue relief. <P>SOLUTION: The functional food having anti-fatigue effect is obtained by adding enzyme decomposed substance having the anti-fatigue effect based on finding that the enzyme decomposed substance produced by treating animal muscle (skeletal muscle) with endo-form protease and/or exo-form protease has high anti-fatigue activity. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

  The present invention relates to an anti-fatigue agent or a functional food having anti-fatigue action, more specifically, an anti-fatigue agent or anti-fatigue action containing a peptide having anti-fatigue activity obtained by degrading pork red meat with protease. It relates to functional foods.

  In today's society, many people have problems related to fatigue, such as excessive work load, shortening of sleep time, stress due to changes in the surrounding environment, etc. Fatigue is defined as "a diminished state of physical function with a unique pathological discomfort caused by excessive physical and mental activity and a desire for rest". There are central fatigue and peripheral fatigue. Central fatigue is a mental fatigue of the cranial nervous system and is mainly caused by stress. Peripheral fatigue, on the other hand, is caused by muscle exercise, and is thought to be mainly caused by energy depletion, accumulation of fatigue substances, and internal environment malfunction.

  Recently, the presence of active oxygen has been attracting attention as a cause of fatigue, and it has come to be regarded as an important fatigue substance. The excessive inflammatory reaction associated with increased oxygen flow to active muscle tissue and muscle stretch during exercise is It is said to promote the generation of active oxygen in the living body. In addition, because the cell membranes that make up muscles are rich in lipids, they tend to produce lipid peroxides, which can damage muscles and lead to contractile dysfunction. It is said that the phenomenon becomes conscious of fatigue.

  In general, damage in vivo due to active oxygen can be reduced or eliminated by antioxidants, and antioxidants contained in foods include vitamin C, vitamin E, β-carotene, and the like. Data suggesting that substance intake is effective in relieving fatigue has also been shown, but no theory has been reached.

  On the other hand, peptide nutrition with a peptide chain length of 2-10, obtained by hydrolyzing protein such as soybean protein with protease, used to recover physical exhaustion due to intense sports and labor and to recover early physical strength after surgery When receiving agents or food compositions (see, for example, Patent Documents 1 and 2) and passive recovery (for some time in ships, vehicles, etc.) as well as recovery from fatigue in active exercise Dipeptides having a molecular weight of 700 or more and a peptide having a molecular weight of 700 or more and a free amino acid content of 5 wt% or less obtained by hydrolyzing isolated soybean protein, casein, ovalbumin, etc. A low molecular weight peptide composition comprising a low molecular weight peptide composition having an average molecular weight of 200-550 based on a tripeptide Fatigue agent with (e.g., see Patent Document 3) are known.

  In addition, the use of beef (including internal organs) from one or more of proteolytic enzymes derived from microorganisms, animals and plants to increase the physical strength and fatigue recovery from a water-soluble beef peptide mixture obtained by enzyme treatment (For example, refer to Patent Document 4), or a meat processing waste liquid and a meat extract, which is a concentrate thereof, are electrodialyzed in an acidic to neutral region, and are composed of a water-soluble peptide and an effective water-soluble component. -Carnitine and histidine-related dipeptides (carnosine, anserine, barenine) and taurine-containing material for enhancing physical fitness and fatigue and foods using the same (for example, see Patent Document 5) and imidazole dipeptides (anserine, carnosine, and barenine) And / or its salt as an active ingredient to improve exercise capacity and anti-fatigue Known anti-fatigue compositions having fruit (for example, see Patent Document 6) and meat products having an anti-fatigue effect by adding carnitine and creatine, which are muscle-derived components, to meat products (for example, see Patent Document 7) ing. Furthermore, an antioxidative peptide such as carnosine obtained by pulverizing, extracting and separating livestock products or livestock by-products, and a functional food or beverage containing the antioxidative peptide as an active ingredient (for example, see Patent Document 8) are known. It has been.

JP-A 63-287462 JP-A-6-14747 JP-A-4-149138 Japanese Patent No. 3563892 JP 2001-46021 A JP 2002-173442 A JP 2003-135033 A JP 2003-267992 A

  An object of the present invention is to obtain an effective solution for fatigue-related problems that many modern people suffer from. Specifically, the anti-fatigue agent is effective and effective in preventing fatigue and recovering from fatigue. And to provide functional food.

  Conventionally, meat is effective for recovery from fatigue, water-soluble peptide obtained by treating beef (including internal) with proteolytic enzyme is used as an anti-fatigue agent, L-carnitine and histidine-related in meat Although it has been known that dipeptides (carnosine, anserine, and valenin) and taurine are used as anti-fatigue agents, the present inventors have further studied and intensively investigated peptides that act more effectively as anti-fatigue agents. As a result, the peptide-containing fraction obtained by treating animal muscle (skeletal muscle) with a protease was found to have high anti-fatigue activity, and the present invention was completed.

  That is, the present invention includes [1] a functional food material or functional food and drink having an anti-fatigue effect, characterized by containing a composition for proteolytic treatment of pulverized and / or ground products of meat, and [2] meat. The anti-fatigue agent according to the above [1], wherein the pulverized and / or ground product is a pulverized and / or ground product of meat from which fat and connective tissue in the meat are removed as much as possible, [3] A functional food material or a functional food or drink having an anti-fatigue effect as described in [1] or [2] above, which is obtained by adding a protease to an enzyme treatment; [4] an enzyme The functional food material having an anti-fatigue effect according to the above [3], wherein the treatment is a treatment of inactivating the protease by heat treatment after the endo-type protease and / or exo-type protease is acted on Or function On food and drink.

  The present invention also provides [5] after crushing and / or grinding the meat, or when crushing and / or grinding the meat, adding protease to perform enzyme treatment, crushing the meat and / or protein of the ground product. A method for producing a functional food material or a functional food or drink product having an anti-fatigue effect characterized by obtaining a decomposition treatment composition, or [6] enzyme treatment caused endo-type protease and / or exo-type protease to act The method for producing a functional food material or functional food or drink having an anti-fatigue effect as described in [5] above, wherein the protease is inactivated by heat treatment, or [7] in meat The functional food material or function having an anti-fatigue effect according to any one of [5] or [6] above, wherein the meat from which fat and connective tissue have been removed as much as possible is pulverized and / or ground. sex Food process for the preparation of.

  ADVANTAGE OF THE INVENTION According to this invention, the anti-fatigue agent and functional food which are effective in the recovery | restoration and prevention of the fatigue which many people of the modern age suffer from using the muscle (skeletal muscle) of an animal as a raw material can be provided.

  The anti-fatigue agent of the present invention is not particularly limited as long as it is an anti-fatigue agent containing a peptide-containing fraction obtained by treating animal muscle (skeletal muscle) with a protease as an active ingredient. Especially as long as it is a food (food material) containing a peptide-containing fraction obtained by treating animal muscle (skeletal muscle) with a protease and having anti-fatigue activity on the package or instructions It is not limited. Pigs, cows, chickens and the like are suitable for the skeletal muscles to be used, but are not limited to these breeds. Moreover, the site | part and state of a muscle are not specifically limited. Further, the protease used for decomposing animal muscles (skeletal muscles) is not limited to a specific one, and any type can be used as long as it appropriately decomposes animal muscles (skeletal muscles).

  In the protease treatment, the crushed and / or ground meat is enzymatically treated with endo-type protease or exo-type protease. The endo-type protease and exo-type protease are not particularly limited, and commercially available ones can be used. However, proteases having an optimum working temperature around 50 ° C., and the meat itself has a slightly acidic pH. Proteases having an optimum pH on the weakly acidic side are preferable, and these can be used singly or in combination. When these enzymes are used in combination, they may be used simultaneously or sequentially. Also good. The enzyme treatment conditions, that is, the type of protease, the action temperature, the action pH, the reaction time, the amount used, etc. are not particularly limited as long as the proteolytic treatment composition having anti-fatigue activity is obtained. Is preferably 1 to 5 hours, particularly about 2 to 4 hours, since the mild reaction is preferred, and the reaction temperature is preferably 45 to 55 ° C.

  The heat treatment applied after the protease treatment inactivates the protease. Examples of the heat treatment include water dispersibility of the gel-like material after the heat treatment, sterilization of the gel-like material after the heat treatment, and heat treatment efficiency. In view of the above, a heat treatment at 85 to 95 ° C. for 120 to 45 minutes is preferable, and a heat treatment at 90 ° C. for 1 hour is particularly preferable. By such heat treatment, the protease treatment composition after the protease treatment usually exhibits a gel form. When water is added to this gel-like proteolytic treatment composition, for example, when water is added in the range of 90 to 40 parts by weight with respect to 10 to 60 parts by weight of the gel-like product after heat treatment, uniform suspension A dispersion can be obtained. In addition, since the meat processing material may be mixed with foreign matters such as hair, cartilage, bone, etc., it is preferable to subject the gel-like aqueous suspension after the heat treatment to a filtration treatment.

  Protease-treated animal muscle (skeletal muscle) protease degradation products contain peptides that have anti-fatigue effects, and there are no coexisting substances that greatly inhibit the effects, so they can be used as foods (food materials) as they are. it can. However, when it is necessary to increase the effectiveness, it is desirable to concentrate and purify the peptide having anti-fatigue activity. For example, a method of concentration by freeze-drying or fractionation by liquid chromatography can be mentioned. .

  The anti-fatigue agent containing a peptide or a salt thereof contained in the protease degradation product of animal muscle (skeletal muscle) described above as an active ingredient is usually administered (ingested) to humans orally. The amount necessary to show the anti-fatigue effect is about 50 to 100 mg / kg / day in terms of decomposition products. When the antioxidant of the present invention is orally administered (taken), excipients (lactose, starch, etc.), binders (syrup, gelatin, etc.), additives ( Nutritional additives such as vitamins and minerals, sweetening agents, flavoring agents, flavoring agents such as pigments, appearance improvers, etc.) can be used.

  The amount of intake necessary for the functional food containing the peptide or salt thereof contained in the protease degradation product of the animal muscle (skeletal muscle) described above as an active ingredient to have an anti-fatigue effect is, in terms of degradation product, It is about 50 to 100 mg / kg / day. Adding or processing (mixing, heating, etc.) the peptide or its salt as an active ingredient or a peptide-containing fraction into an appropriate food or food material does not cause any inconvenience.

  There are no particular restrictions on the types of such foods and food materials, such as yogurt, drink yogurt, juice, milk, soy milk, liquor, coffee, tea, sencha, oolong tea, sports drinks, pudding, cookies, etc. Baked confectionery such as bread, cake, rice cracker, Japanese confectionery such as sheep crab, bread and confectionery such as frozen confectionery, jelly and chewing gum, noodles such as udon and soba, fish paste products such as kamaboko, ham and fish sausage, miso Examples include seasonings such as soy sauce, dressing, mayonnaise and sweeteners, dairy products such as cheese and butter, various prepared dishes such as tofu, konjac, other boiled fish, dumplings, croquettes, salads, honey, royal jelly, etc. it can.

  Hereinafter, the present invention will be described with reference to examples. The following examples are given to illustrate the present invention and are not intended to limit the present invention.

(Method of preparing protease degradation product of animal muscle (skeletal muscle))
A protease degradation product of animal muscle (skeletal muscle) was prepared from porcine skeletal muscle. That is, 1 g of sodium chloride and 0.04 g of sodium ascorbate are added to porcine skeletal muscle (biceps femoris) that has been minced after removing fat and connective tissue as much as possible, and endo-type protease "Amano S" ) And exo-type protease “Amano P3G” (manufactured by Amano Pharmaceutical Co., Ltd.) were added in an amount of 50 ppm each, and the mixture was crushed with good mixing with a silent cutter. The crushed meat was vacuum-packed and subjected to an enzyme reaction at 50 ° C. for 3 hours, and then heated at 90 ° C. for 1 hour to inactivate the protease to obtain a protease degradation product.

(Measurement method of antioxidant activity)
Antioxidant activity was measured using a method in which superoxide ions were quantified by chemiluminescence. First, hypoxanthine was reacted with xanthine oxidase in the presence of a measurement sample (antioxidant or control water) to generate superoxide ions. The produced superoxide ion was allowed to react with 2-methyl-6-p-methoxyphenylethynylimidazopyrazinone (MPEC, manufactured by Ato Co., Ltd.) as a luminescence reagent, and the amount of luminescence was measured with a luminescence sensor. Antioxidant activity was calculated by the following formula.
Antioxidant activity (%) = (Measured value of control? Measured value of sample) ÷ Measured value of control × 100

(Measurement method of anti-fatigue effect)
The anti-fatigue effect was determined by the extent of the running time when the mice to which the sample (protease degradation product) was orally administered were forcibly run (compared to the control group to which water and non-enzyme-treated pork homogenate were orally administered). Preliminary breeding of ICR male mice up to 4 weeks of age, preliminarily running to familiarize them with the forced exercise running device (treadmill MK-680S, made by Muromachi Kikai Co., Ltd.) from 4 weeks of age to determine the anti-fatigue effect Used 5 to 6 weeks old. The forced running was performed with the inclination of the treadmill running surface being 15 degrees and the belt speed being 20 m / min.

  Before the sample was administered, the mice were forcibly run for 4 hours on a treadmill to bring them to a certain degree of fatigue, and then the sample solution was orally administered using a stainless gastric sonde. The sample volume administered orally was 0.2 ml. The sample concentration was 10 mg / ml (administration dry matter weight per mouse: 2 mg). For comparison, 0.2 ml of non-enzyme-treated pork homogenate was similarly administered at 10 mg / ml (administration dry matter weight per mouse: 2 mg). In addition, 0.2 ml of water was administered to the control group. Forced running was resumed 15 minutes after the aforementioned 4 hours of forced running (including the time required for oral administration of the sample), and the running time of the mice was measured. The time when the mouse stopped running and kept in contact with the electrode at the rear of the treadmill for 5 seconds or more was regarded as the end of running. The electrode was set at a voltage of 100 V so that the mouse continued to run to the limit of fatigue (since the mouse dislikes contact with the electrode, it continued to run until the fatigue level reached the limit).

(Antioxidant activity and anti-fatigue effect of protease degradation products)
Antioxidant activity (superoxide ion scavenging ability) of a protease degradation product of pig skeletal muscle (sample concentration in the reaction solution: 0.4 mg / mL) was measured. In addition, unenzyme-treated pork was used as a comparative control. As the result shown in FIG. 1, the protease degradation product showed a strong antioxidant activity compared to the non-enzyme-treated pork.

  Proteolytic degradation products showing strong antioxidant activity were orally administered to mice (n = 14-17) in an amount of 2 mg / mouse, and the anti-fatigue effect was examined by forced running test. As shown in the results of FIG. 2, oral administration of the protease degradation product significantly increased the running time of the mice. This running time is about 5 times that of the control group to which water was administered.

It is the graph which showed the result of having measured the antioxidant activity (superoxide ion scavenging ability) of protease degradation product. It is the graph which compared the anti-fatigue effect (extension of running time) at the time of orally administering a protease degradation product etc. to a mouse.

Claims (7)

A functional food material or a functional food or drink having an anti-fatigue effect, comprising a composition for proteolytic treatment of pulverized meat and / or ground product. The function having an anti-fatigue effect according to claim 1, wherein the ground and / or ground product of meat is a ground and / or ground product of meat from which fat and connective tissue in the meat are removed as much as possible. Food or functional food. A proteolytic treatment composition for pulverized and / or ground meat is obtained by enzymatic treatment after addition of protease after pulverizing and / or grinding meat or during pulverizing and / or grinding meat. A functional food material or a functional food or drink having an anti-fatigue effect according to claim 1 or 2. The functional food material having an anti-fatigue effect according to claim 3, wherein the enzyme treatment is a treatment in which the protease is deactivated by heat treatment after the endo-type protease and / or exo-type protease is allowed to act. Or functional food and drink. After pulverizing and / or grinding the meat, or when pulverizing and / or grinding the meat, protease is added to perform enzyme treatment to obtain a composition for proteolytic treatment of the pulverized meat and / or the ground product. A method for producing a functional food material or functional food or drink having a characteristic anti-fatigue effect. 6. The functional food material having an anti-fatigue effect according to claim 5, wherein the enzyme treatment is a treatment for inactivating the protease by heat treatment after allowing the endo-type protease and / or exo-type protease to act. Or the manufacturing method of functional food-drinks. 7. A functional food material having anti-fatigue effect or functionality according to any one of claims 5 to 6, wherein the meat from which fat and connective tissue in the meat are removed as much as possible is pulverized and / or ground. A method for producing food and drink.