JP6136295B2 - Muscle enhancer - Google Patents

Description

  The present invention relates to a muscle enhancer.

In recent years, the demand for animal protein has increased in Japan with changes in dietary habits. For example, breeding selection of dedicated varieties for meat, improvement of nutrition and feed, etc. have been carried out in poultry, and as a result, chicken, which is a high quality animal protein, is stably supplied. However, due to the pursuit of productivity improvement so far, in addition to problems such as fatty liver caused by increased intra-abdominal fat in meat chickens, fat is removed during the broiler treatment process, most of which is discarded. Therefore, there is a problem that increase in intraperitoneal fat leads to loss of energy in the feed.
Thus, in order to obtain meat of good quality livestock products, it is desired to grow individuals with a lot of muscles and without excess fat. Furthermore, it is desirable from the viewpoint of cost effectiveness if the feed can be efficiently replaced with muscle.
In addition, lifestyle-related diseases such as metabolic syndrome, obesity, diabetes, and hyperlipidemia have become a problem in recent society. However, if efficient production of low-fat and high-protein meat can be achieved, these measures can be taken. It can contribute to the maintenance of people's health, which in turn leads to an improvement in QOL.

  Regarding muscle augmentation, there is a technique using wheat germ (Patent Document 1), but there is no knowledge so far regarding the effects of carbohydrates such as disaccharides on muscle augmentation.

  On the other hand, the treatment of enzyme in palm kernel meal, copra meal, etc., and β-1,4-mannobiose content increased to 10% or more is added to the feed to suppress Salmonella colonization in the animal intestine. It is known to have a sterilizing effect (salmonella colonization suppression effect) excreted outside the body, and various techniques using this effect have been proposed (Patent Document 2).

  Similarly, the enzyme efficiency of palm kernel meal, copra meal, etc. is increased to 3% or more of β-1,4-mannobiose content, and the feed efficiency is improved by adding it to feed for pig farming or poultry farming. It is known (Patent Document 3)

JP 2012-77010 A Japanese Patent No. 4570959 WO2011 / 027753

Food and Development Vol 35, No. 2, p54-56, 2000

  However, this Patent Document 3 is intended to improve productivity and, consequently, economy by increasing the efficiency of increasing the total weight of livestock animals in feed applications, and does not mention any effect on individual skeletal muscles. . Therefore, an object of the present invention is to provide a muscle enhancer that exhibits effects such as enhancing muscles of livestock animals.

  As a result of intensive studies to solve the above problems, the present inventors have gained superiority in muscles of livestock animals by ingesting a muscle enhancer containing 20% by weight or more of β-1,4-mannobiose. The present invention has been completed.

That is, the present invention
(1) A muscle strengthening agent characterized by containing β-1,4-mannobiose in an amount of 20% by weight or more.
(2) The muscle strengthening agent according to (1), wherein the content of β-1,4-mannobiose is 90% by weight or more.
(3) The muscle enhancing agent according to (1) or (2), wherein the muscle is a muscle of the chest.
(4) The muscle enhancer according to (1) to (3), which is obtained by hydrolyzing a polysaccharide contained in a mannan-containing natural product, followed by solid-liquid separation and column purification.
It is.
(5) When the muscle enhancing agent according to (1) to (4) is ingested by a livestock animal, 0.001 g or more per day is consumed per 1 kg of the body weight of the livestock animal. Enhancement method.

  The muscle enhancer containing high-purity β-1,4-mannobiose of the present invention increases ribosomal capacity, which is an index of protein synthesis, and suppresses myostatin, which is a factor that inhibits protein synthesis. It is possible to increase the muscle weight of livestock animals such as pigs, that is, to strengthen the muscles, and the effect on the muscles of the chest is particularly high.

FIG. 1 is a graph showing breast weight per body weight after ingestion of β-1,4-mannobiose or control feed in a breeding test using chickens. FIG. 2 is a diagram showing ribosomal capacity, which is an index of muscle protein metabolism of breast meat after ingestion of β-1,4-mannobiose or control feed in a breeding test using chickens. FIG. 3 shows the expression level of myostatin mRNA, which is a regulator of suppression of differentiation and development of myoblasts of breast muscle after ingestion of β-1,4-mannobiose or control diet in a breeding test using chickens. FIG.

(Muscle enhancer)
The muscle strengthening agent according to the present invention contains β-1,4-mannobiose (hereinafter, also simply referred to as mannobiose) at a high content. The content of β-1,4-mannobiose is 20% by weight or more, preferably 90% by weight or more.

(Β-1,4-mannobiose)
In the present invention, β-1,4-mannobiose is formed by bonding two mannose molecules to β-1,4-glycoside. The β-1,4-mannobiose used in the present invention can be obtained, for example, by a method of synthesizing from mannose or a method of decomposing β-1,4-mannan (hereinafter also simply referred to as mannan).

  The method for decomposing β-1,4-mannan is more preferable in terms of the raw material resources and reaction efficiency, and β-1,4-mannobiose can be obtained more easily. In this method, for example, mannan-degrading enzyme acts on mannan-containing natural products such as palm kernel meal, copra meal, coffee bean meal, sesame meal, guar gum, locust bean gum, etc., which are rich in mannan, or mannan extracted from these natural products. Thus, β-1,4-mannobiose can be obtained.

  When the raw material containing mannan polysaccharide has a large particle size, it is preferably used after it has been granulated or powdered in an appropriate size by a crusher or pulverizer. The fat component in the raw material is preferably used after degreasing in advance using an organic solvent such as hexane or ethanol.

  The raw material is preferably treated with an alkaline solution to selectively solubilize and remove components other than the mannan polysaccharide, which is an impurity, before use. That is, for example, by directly suspending the raw material in a solution adjusted to be alkaline in advance, or by suspending the raw material in water and adjusting the suspension to be alkaline, the solution is allowed to stand by stirring or stirring for 5 to 120 minutes. Perform solution treatment. Next, if necessary, an acid such as hydrochloric acid, sulfuric acid, nitric acid, citric acid or propionic acid is added to the suspension to neutralize it to the original suspension pH, for example, about pH 5 to 7, and then centrifuged. Solid-liquid separation is performed by separation or filtration, and the residue is recovered. Moreover, it is good to perform 1 to several times of water washing as needed. That is, after adding water to the obtained residue, stirring and standing, the residue is recovered again by solid-liquid separation such as centrifugation or filtration. The washing process may be repeated as necessary.

  Next, the raw material of the mannan polysaccharide subjected to the above-described treatment is hydrolyzed to produce mannobiose which is a disaccharide. In order to produce mannobiose with higher purity, it is preferable to use a mannan degrading enzyme. The mannan degrading enzyme used in this method may be any mannanase, mannosidase, hemicellulase, etc., as long as it has an activity of degrading mannan to produce mannobiose, and is commercially available from Aspergillus niger. (For example, hemicellulase GM “Amano” (manufactured by Amano Pharmaceutical Co., Ltd.), Sumiteam ACH (manufactured by Shin Nippon Chemical Industry Co., Ltd.), cellulosin GM5 (manufactured by Hankyu Bioindustry Co., Ltd.), etc. can be preferably used. In addition to those commercially available as xylanase and cellulase, those having the hydrolysis activity can also be used, for example, cellulase Y-NC (manufactured by Yakult Pharmaceutical Co., Ltd.), in particular, mannosidase ( (exo type) activity is low, mannanase (endo type) activity is low High hemicellulase GM “Amano” (manufactured by Amano Pharmaceutical Co., Ltd.) and Sumiteam ACH (manufactured by Shin Nippon Chemical Industry Co., Ltd.) are preferable because they can suppress the production of mannose and can produce a large amount of mannobiose.

  Furthermore, in this method, the mannan degrading enzyme is allowed to act on mannan-containing natural products or mannan extracted therefrom as an enzyme solution dissolved or dispersed in water. In order to perform an efficient reaction in the case of using a mannan-containing natural product, it is important to adjust moisture in a reaction system including the mannan-containing natural product, a mannan-degrading enzyme, and water. The amount of water added for moisture adjustment is preferably 100 to 10000 parts by weight, more preferably 500 to 1000 parts by weight with respect to 100 parts by weight of mannan. By setting the amount of water to be in such a range, the mannan fibers can be sufficiently swollen in the presence of sufficient moisture, and the enzyme solution can be easily contacted.

  As conditions for performing the enzyme reaction, there is no particular problem as long as it is a normal enzyme reaction condition, and an optimum condition may be selected for the enzyme to be used. The reaction temperature is desirably a temperature range in which the enzyme is not inactivated and the microorganisms do not grow, and is preferably 40 to 80 ° C, more preferably 50 to 70 ° C. The pH in the reaction is preferably carried out under the optimum conditions of the enzyme to be used, but in the same way as the reaction temperature, an acidic or weakly acidic range of pH 2 to 6 is desirable in order to prevent spoilage by microorganisms. Although the reaction time depends on the amount of enzyme used, it is preferably 3 to 48 hours for work convenience. After hydrolysis, solid-liquid separation is performed, and the supernatant is recovered.

  Since the supernatant obtained above contains 40% by weight or more of mannobiose per dry matter weight, the supernatant liquid is concentrated as it is or concentrated by an evaporator or the like, and further lyophilized from these liquids to enhance muscle. It can be used as an agent. However, the above supernatant liquid contains up to about 60% of mannose, mannotriose, mannohexaose, other manno-oligosaccharides, and also glucose, galactose, sucrose and other oligosaccharides. Therefore, in order to separate these sugars and increase the purity of mannobiose, it is preferable to purify the column after solid-liquid separation. As the method, for example, using a column using a hydrophobic resin, a fraction collector, etc. And a method of fractionating only mannobiose with high purity can be employed. Examples of the hydrophobic resin include HP-20 (Mitsubishi Chemical Corporation), Bio-Gel P (BIO-RAD) and the like. As a result, 90% by weight or more of mannobiose per dry weight can be fractionated. By concentrating the mannobiose fraction with an evaporator or the like and further freeze-drying it, a muscle enhancer containing 90% by weight or more of mannobiose can be obtained.

  The muscle-enhancing agent of the present invention exerts the effect of increasing various muscles of livestock animals such as chickens and pigs, that is, the effect of strengthening muscles.

  The muscle strengthening agent of the present invention can be used not only for broiler species but also for various brand chickens and chickens whose selling points are differences in meat quality, taste and texture. It can also be used for pigs, cows, sheep, horses and the like.

  When the muscle strengthening agent of the present invention is used for livestock animals such as chickens and pigs, it is preferably added such that the blending amount in the feed is 0.005 wt% to 0.15 wt% as mannobiose. Is suitably from 0.006% to 0.1% by weight, most preferably from 0.007% to 0.05% by weight. Such feed can strengthen muscles by feeding livestock in solid or liquid form.

  In addition, the amount of the muscle-enhancing agent of the present invention ingested by livestock animals such as chickens and pigs is preferably 0.001 g or more, more preferably 0.005 g to 0.15 g per day with respect to 1 kg of the weight of livestock animals. More preferably, 0.006 g to 0.1 g, and most preferably 0.007 g to 0.05 g is appropriate.

  Hereinafter, more specific embodiments of the present invention will be described. In the examples, “parts” and “%” represent “parts by weight” and “% by weight” unless otherwise specified.

Example 1
The coconut powder obtained from the Philippines was mixed with hexane twice as much by weight as the weight ratio, and then filtered to recover the residue. This operation was further repeated twice to obtain a defatted coconut powder having a residual oil content of 0.1%. To 500 g of this defatted coconut powder, 7500 g of distilled water and 10 g of commercially available mannanase “Sumiteam ACH-L” (manufactured by Shin Nippon Chemical Co., Ltd.) are added, under conditions of pH 5, reaction temperature 50 ° C. and reaction time 20 hours. Hydrolysis reaction was performed. After the reaction, centrifugation (10,000 rpm, 20 minutes) was performed, and the supernatant was collected. After concentration by an evaporator, freeze drying was performed. The recovered amount of the lyophilized product was 75 g. Furthermore, a fraction collector was attached to a glass column (diameter: about 10 cm × length: about 100 cm) that was dissolved in water and packed in advance with a hydrophobic resin Bio-Gel P (BIO-RAD). After being dissolved in water and applied to the column, 6 L of water was passed at a flow rate of 5 mL / min for fractionation.
After the total sugar analysis of the sample of each column, the sugar analysis is performed by the ion chromatography ICS-3000 system of Dionex, the fraction containing mannobiose is collected, freeze-dried, and containing 99% or more of mannobiose 3 g of a muscle enhancing agent was obtained. This column operation was repeated 12 times to obtain about 40 g of a muscle enhancer containing 99% or more of mannobiose.

(Example 2)
To copramir (produced by Fuji Oil Co., Ltd.), twice the weight of hexane was added and mixed, followed by filtration to recover the residue. This operation was further repeated twice to obtain defatted copra meal having a residual oil content of 0.1%. To 500 g of this defatted copra meal, 7500 g of distilled water and 10 g of commercially available mannanase “Sumiteam ACH-L” (manufactured by Shin Nippon Chemical Co., Ltd.) are added, and water is added under the conditions of pH 5, reaction temperature 50 ° C. and reaction time 20 hours. A decomposition reaction was carried out. After the reaction, centrifugation (10,000 rpm, 20 minutes) was performed, and the supernatant was collected. After concentration by an evaporator, freeze drying was performed. The recovered amount of the lyophilized product was 100 g. Furthermore, a fraction collector was attached to a glass column (diameter: about 10 cm × length: about 100 cm) that was dissolved in water and packed in advance with a hydrophobic resin Bio-Gel P (BIO-RAD). After being dissolved in water and applied to the column, 6 L of water was passed at a flow rate of 5 mL / min for fractionation.
After the total sugar analysis of the sample of each column, the sugar analysis is performed by the ion chromatography ICS-3000 system of Dionex, the fraction containing mannobiose is collected, freeze-dried, and containing 99% or more of mannobiose 3 g of a muscle enhancing agent was obtained. This column operation was repeated 8 times to obtain about 25 g of a muscle enhancer containing 99% or more of mannobiose.

(Raising test using chicken)
24 broilers (Chunky, Male, Ishii Co., Ltd.) were fed 1 to 7 days of age with a commercial feed made by Nippon Compound Feed Co., Ltd. The test feed (Table 1) containing β-1,4-mannobiose corresponding to 0.01% of the feed or water for 13 days from 8 to 21 days of age was fed. It was measured. The pectoral muscle, which is the main muscle of breast meat, was frozen and then used for analysis of total RNA and protein. The total RNA amount and the protein amount were quantified to calculate the ribosomal capacity (total RNA amount / protein amount ratio), which is an index of muscle protein synthesis. In addition, total RNA was extracted, cDNA was synthesized by reverse transcription, and a complementary DNA of myositatin was amplified using primers as an indicator of skeletal muscle protein metabolism. Ribosomal protein S17 (RPS17) was similarly amplified as an internal standard, and the expression level of these mRNAs was measured using a quantitative device.

(Table 1) Test feed composition table

  As a result of this study, administration of mannobiose significantly increased the weight of breast meat per chicken body weight (FIG. 1). Moreover, in the measurement results of muscle protein metabolism factor of breast meat, ribosomal capacity (total RNA amount / protein amount ratio), which is an index of protein synthesis ability, was significantly increased, and the increase in muscle was due to induction of protein synthesis Was suggested (FIG. 2). From this, this agent can also be used as a muscle protein synthesis promoter.

  In addition, the measured value of myostatin mRNA, which is a regulatory regulator of muscle myoblast differentiation and neoplasia, is significantly reduced by administration of mannobiose, suppressing the inhibition of myoblast differentiation, that is, promoting myoblast differentiation. (Fig. 3). From this, this agent can also be utilized as a myoblast differentiation suppression reducing agent.

From the results of this study, it was found that the addition of β-1,4-mannobiose reinforces chest muscles and that the mechanism is due to an increase in ribosomal capacity, which is an indicator of protein synthesis ability. That is, it was suggested that β-1,4-mannobiose increases protein synthesis in the breast muscle.
It was also suggested that the decrease in myostatin promoted differentiation and differentiation of myoblasts.
Therefore, it has been shown by the present invention that mannobiose can be a useful muscle enhancer that can increase muscle weight based on protein biosynthesis and promotion of muscle differentiation at the cellular level.

Claims (5)

A muscle strengthening agent containing 20% by weight or more of β-1,4-mannobiose. The muscle strengthening agent according to claim 1, wherein the content of β-1,4-mannobiose is 90% by weight or more. The muscle strengthening agent according to claim 1 or 2, wherein the muscle is chest muscle. The muscle strengthening agent according to claims 1 to 3, which is obtained by hydrolyzing a polysaccharide contained in a mannan-containing natural product, followed by solid-liquid separation and column purification. A chicken muscle strengthening method, wherein when the chicken is ingested with the muscle strengthening agent according to any one of claims 1 to 4, 0.001 g or more is ingested per day per kg of chicken body weight.

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