Composition for increasing muscle mass

Abstract

[Problem] Provided is a composition which can be easily ingested and which can increase muscle mass. [Solution] This composition for increasing muscle mass contains (A) guar gum degradation products with an average molecular weight of 1.8 × 103 to 2.0 × 105 and with at least 70 mass% thereof being within this range, and (B) a guar bean protein, the viscosity of a 1 mass% water solution of the composition, measured at 25°C and 60 rpm using a B-type viscometer, is less than or equal to 50 mPa*s, the guar gum degradation products are contained in endosperm derived from guar, and the composition is obtained by using microbe-derived β-mannanase to hydrolyze and reduce the molecular weight of galactomannan polysaccharide that has a galactose and mannose content ratio (galactose:mannose) in the range 1:1.5-1:2.1, wherein the dietary fiber content, specified by enzymatic-HPLC method, is at least 70 mass%, the content of oligosaccharides of the guar gum degradation products is less than or equal to 15 mass%; of the amino acid compositions in the contained proteins, (glutamic acid + glutamine + aspartic acid + asparagine) ≥ 100 g / 100g, (cystine + tyrosine + serine + threonine) ≥ 40mg / 100g, all amino acids ≥ 150mg / 100g, and the ratio (A) / (B) is less than or equal 1000.

Description

[Technical Field] 【0001】 This invention relates to a composition for increasing muscle mass. [Background technology] 【0002】 Peroxisome proliferator-activated receptor γ coactivator-1 (hereinafter referred to as "PGC-1") is a molecule identified as a transcriptional coactivator that binds to the transcription factor PPARγ. It is known to be involved in enhancing mitochondrial biosynthesis and maintaining its function, and to regulate gene expression related to energy production. PGC-1 is thought to play a useful role in processes such as skeletal muscle transformation through exercise training, and research and development are underway to find substances that can promote its biosynthesis in the body (Patent Document 1). On the other hand, valine, leucine, and isoleucine are known as branched-chain amino acids, which have branched side chains. Branched-chain amino acids are essential amino acids that cannot be produced in the body, and are known to be endurance amino acids that act as energy in muscles during exercise (Patent Document 2). If PGC-1 and branched-chain amino acids can be effectively increased, they can have beneficial effects on muscle metabolism, including muscle mass increase, inhibition of muscle mass loss, anti-sarcopenia, anti-frailty, anti-metabolic syndrome based on improved metabolism, anti-fatigue, and anti-aging effects. Therefore, they could be beneficial not only for athletes but also for the elderly, the sick, livestock, and pets. [Prior art documents] [Patent Documents] 【0003】 [Patent Document 1] Republished Gazette No. 2018-207741 [Patent Document 2] Special Publication No. 2020-528402 [Disclosure of the Invention] [Problems that the invention aims to solve] 【0004】 However, Patent Document 1 uses a specific strain of Enterococcus faecium R30, which lacks versatility. Furthermore, Patent Document 2 involves ingesting multiple substances containing branched-chain amino acids, requiring the intake of large quantities of substances, which places a burden on the person taking the substance. This invention was made to solve the above problems and aims to provide a composition that can be easily ingested and can increase muscle mass. [Means for solving the problem] 【0005】 As a result of diligent research to solve the above problems, we discovered that a composition containing a specific galactomannan hydrolysate and guar bean protein has a muscle mass-increasing effect, and thus we have essentially completed the present invention. Thus, the muscle mass increasing composition according to the present invention has (A) an average molecular weight of 1.8 × 10 3 ~2.0×10 5 (B) Guar gum hydrolysate containing 70% or more by mass of materials within the average molecular weight range and guar bean protein, and having a viscosity of 50 mPa·s or less for a 1% aqueous solution when measured at 25°C and 60 rpm using a B-type viscometer, wherein the guar gum hydrolysate is obtained by hydrolyzing galactomannan polysaccharide, which is contained in the endosperm of guar and has a galactose-to-mannose content ratio (galactose:mannose) in the range of 1:1.5 to 1:2.1, using microbially derived β-mannanase to reduce its molecular weight, and having a dietary fiber content of at least 70% by mass as defined by the enzyme-HPLC method, and an oligosaccharide content of guar gum hydrolysate of 15% by mass or less, and among the amino acids in the contained protein, (glutamic acid + glutamine + aspartic acid + asparagine) ≥ 100 mg / 100 g, (cystine + tyrosine + serine + threonine) ≥ 40 mg / 100 g, and total amino acids ≥ 150 The concentration is mg / 100g, and the ratio of (A) / (B) is 1000 or less. In this composition, the ratio of (A) / (B) is 25 to 1000, preferably 100 to 600, and more preferably 200 to 500. Within this range, a sufficient muscle mass increase effect can be obtained. In this case, it is preferable that the above composition has the effect of increasing PGC-1 expression and promoting branched-chain amino acid production. 【0006】 Food and beverage products according to another invention are characterized by containing the above-mentioned composition for increasing muscle mass. Furthermore, it is preferable that the composition has the effect of increasing PGC-1 expression, promoting branched-chain amino acid production, suppressing muscle mass loss, anti-sarcopenia, anti-frailty, anti-metabolic syndrome, anti-fatigue, and anti-aging effects. Furthermore, a pharmaceutical product relating to another invention is characterized by containing the above-mentioned composition for increasing muscle mass. Preferably, it also has the effects of increasing PGC-1 expression, promoting branched-chain amino acid production, suppressing muscle mass loss, anti-sarcopenia, anti-frailty, anti-metabolic syndrome, anti-fatigue, and anti-aging. The food and beverages in this invention include food and beverages, and examples include nutritional supplements, health foods, foods for specified health uses, foods with functional claims, dietary therapy foods, comprehensive health foods, supplements, tea beverages, coffee beverages, juices, soft drinks, drinks, cooked rice, bread, noodles, dairy products, egg products, processed seafood and livestock products, confectionery, oils and fats and processed oils and fats products, seasonings, and prepared foods. Pharmaceuticals include pharmaceuticals or quasi-drugs, and are preferably oral or enteral preparations, and can be in the form of liquids, tablets, granules, pills, syrups, etc. 【0007】 In this invention, "feed" refers to food intended for consumption by organisms other than humans, and its form is not particularly limited. While there are no particular limitations on the organisms to which the feed can be applied, examples include farmed animals and pet animals. Examples of farmed animals include livestock such as horses, cattle, pigs, sheep, goats, camels, and llamas; laboratory animals such as mice, rats, guinea pigs, and rabbits; and poultry such as chickens, ducks, turkeys, and ostriches. Examples of pet animals include dogs and cats. 【Advantages of the Invention】 【0008】 According to the present invention, it is possible to provide a composition or the like that can be easily ingested and promotes an increase in muscle mass. 【Brief Description of the Drawings】 【0009】 [Figure 1] It is a graph showing the measurement results of muscle weights of the high-fat diet group and the PHGG group in a PHGG ingestion test using lifestyle disease model mice. [Figure 2] It is a graph showing the measurement results of LDL-C of the high-fat diet group and the PHGG group. [Figure 3] It is a graph showing the results of examining free amino acids of the control group (C group) and the PHGG group (P group) in a PHGG ingestion test for pigs. (A) Free leucine, (B) Free isoleucine, (C) Free valine, and (D) Results of free branched-chain amino acids are shown respectively. "*" in the graph indicates significance at a risk rate of 5% (p < 0.05), and "**" indicates significance at a risk rate of 1% (p < 0.01) (the same in FIG. 4). [Figure 4] It is a graph showing the results of examining free amino acids of the control group (C group) and the PHGG group (P group). (A) Free methionine, (B) Free threonine, and (C) Results of free essential amino acids are shown respectively. 【Modes for Carrying Out the Invention】 【0010】 Next, embodiments of the present invention will be described while referring to the charts. The technical scope of the present invention is not limited by these embodiments, and it can be implemented in various forms without changing the gist of the invention. Guar gum means a water-soluble natural polysaccharide obtained from the endosperm (more precisely, cotyledons) of guar beans, and it is a polysaccharide having a side chain of one molecule of galactose for two linearly bonded molecules of mannose, and the average molecular weight is 2.0×10 5 ~3.0×10 5is the degree. Guar gum is known to have physiological effects such as an effect of suppressing blood glucose level increase, an effect of reducing cholesterol, and an effect of improving bowel movement. In the present invention, the guar gum degradation product means a water-soluble dietary fiber obtained by using beans derived from the annual leguminous plant guar (scientific name: Cyanopsis tetragoloba) used for food in India, Pakistan, etc. as a raw material and hydrolyzing and reducing the molecular weight of the galactomannan polysaccharide contained in its endosperm. The method for hydrolyzing guar gum is not particularly limited, such as an enzymatic decomposition method or an acid decomposition method, but the enzymatic decomposition method is preferable from the viewpoint of easily adjusting the molecular weight of the degradation product. 【0011】 The enzyme used in the enzymatic decomposition method is not particularly limited as long as it is an enzyme that hydrolyzes a mannose straight chain, but it is preferable to use β-mannanase derived from Aspergillus genus bacteria or Rhizopus genus bacteria. The average molecular weight distribution of the guar gum degradation product has an upper limit value of 2×10 5 or less, preferably 1.0×10 5 or less, more preferably 2.5×10 4 or less. The lower limit value of the average molecular weight distribution of the guar gum degradation product is 1.8×10 [[ID=II]] 2 or more, preferably 3.0×10 3 or more. When the average molecular weight exceeds 2×10 5 , the viscosity is too high and it becomes difficult to contain it in food and drink products. The method for measuring the molecular weight distribution is not particularly limited, but for example, polyethylene glycol (average molecular weight: 2×10 2 , 2×10 3 , 2×10 4 and 1×10 5 ) is used as a molecular weight marker, and a method using high performance liquid chromatography method, etc. are available. The guar gum degradation product of the present invention is preferably one in which those within the above average molecular weight range are contained at 70% by mass or more, preferably 80% by mass or more. 【0012】 Galactose is a kind of monosaccharide classified as an aldohexose, and its molecular formula is C6H 12O6 has a molecular weight of 180 (the same as glucose). Its stereochemistry is such that the -OH groups at position 2 (second from the top in the Fischer projection) and position 5 are in the same direction, while those at positions 3 and 4 are in opposite directions. The stereochemistry at position 5 of D-galactose is the same as that of D-glyceraldehyde. Mannose is a type of monosaccharide classified as an aldohexose, with the molecular formula C6H 12 O6 has a molecular weight of 180 (the same as glucose). Its stereochemistry is such that the -OH groups at positions 2 and 3 are in the same direction, while those at positions 4 and 5 are in opposite directions. The stereochemistry at position 5 of D-mannose is the same as that of D-glyceraldehyde. Mannose is not metabolized much in humans and hardly enters the glycolysis pathway when ingested orally. The oligosaccharide content was 6-15% by mass (less than 15% by mass) according to the analysis chart of high-performance liquid chromatography. 【0013】 The amino acid composition of a protein can be measured by known methods, such as HPLC. During the pretreatment for amino acid analysis, asparagine and glutamine are converted to aspartic acid and glutamic acid, respectively, when the protein is hydrolyzed. Therefore, during measurement, asparagine and aspartic acid, and glutamic acid and glutamine, cannot be distinguished. For this reason, they are quantified together as aspartic acid and glutamic acid. The muscle mass increasing composition of the present invention can be taken orally as is or mixed with food, beverages, etc. The dosage for oral intake is not particularly limited, but is 0.5g to 70g per adult per day (preferably 3g to 30g, more preferably 6g to 18g). 【0014】 <Preparation of Guar Gum Hydrolysate (PHGG)> Example 1 900g of water was mixed with 0.1N hydrochloric acid to adjust the pH to 4.5. Then, 0.2g of β-mannanase derived from Aspergillus bacteria (manufactured by Novo Nordisk Bioindustry) and 100g of guar gum powder were added and mixed. This mixture was reacted at 40°C to 45°C for 24 hours. After the reaction, the enzyme was inactivated by heating at 90°C for 15 minutes. The reaction solution was filtered and separated by suction filtration, and the clear solution obtained by removing insoluble matter was concentrated under reduced pressure (using a Yamato evaporator). A solid content of 20% by mass was obtained. This was dried using a spray dryer (manufactured by Okawara Chemical Machinery Co., Ltd.) to obtain 65g of guar gum hydrolysate as powder. Guar gum hydrolysates were dissolved in water to obtain a 0.5 (w / v)% aqueous solution. Polyethylene glycol (average molecular weight: 2 × 10⁻⁶) was used as a molecular weight marker. 2 , 2×10 3 , 2×10 4 and 1 × 10 5 Using high-performance liquid chromatography (column: YMC-Pack Diol-120, detector: differential refractometer), the average molecular weight was determined to be approximately 20,000. The average molecular weight was 1.8 × 10⁶. 3 ~2.0×10 5 It contained more than 85% by mass of that substance. 【0015】 Furthermore, the viscosity of a 1% by mass aqueous solution was measured using a B-type viscometer at 25°C and 60 rpm, and was found to be 8 mPa·s. The ratio of galactose to mannose content (galactose:mannose) was measured to be 1:1.7. The dietary fiber content was measured by enzyme-HPLC and found to be 90% by mass. The oligosaccharide content was 7.3% by mass. Furthermore, an amino acid analysis of the guar bean protein in this example was performed, and the results are shown in Table 1. From these results, the values ​​for (glutamic acid + glutamine + aspartic acid + asparagine) / total amino acids, (glutamic acid + glutamine + aspartic acid + asparagine) / (arginine + lysine + histidine), (cystine + tyrosine) / total amino acids, and (valine + histidine + proline + leucine) / total amino acids were calculated to be 0.47, 5.0, 0.11, and 0.10, respectively. Furthermore, when the ratio of guar gum hydrolysate to guar bean protein was determined, it was found to be 279:1. 【0016】 [Table 1] Due to the characteristics of amino acid analysis, glutamic acid + glutamine (Glu + Gln) is detected as glutamic acid, aspartic acid + asparagine (Asp + Asn) is detected as asparagine, and cysteine ​​(Cys) is detected as cystine. This has been taken into consideration when describing the results in the table. 【0017】 Example 2 900g of water was mixed with 0.1N hydrochloric acid to adjust the pH to 3. Then, 0.15g of β-mannanase derived from Aspergillus bacteria (manufactured by Novo Nordisk Bioindustry) and 100g of guar gum powder were added and mixed. This mixture was reacted at 40°C to 45°C for 24 hours. After the reaction, the enzyme was inactivated by heating at 90°C for 15 minutes. The reaction solution was filtered and separated by suction filtration, and the clear solution obtained by removing insoluble matter was concentrated under reduced pressure (using a Yamato evaporator). A solid content of 20% by mass was obtained. This was dried using a spray dryer (manufactured by Okawara Chemical Machinery Co., Ltd.) to obtain 68g of guar gum hydrolysate as powder. The average molecular weight of the guar gum hydrolysate was determined in the same manner as in Example 1, and was found to be approximately 2.5 × 10⁻⁶. 4 The result was that the average molecular weight was 1.8 × 10⁶. 3 ~2.0×10 4 It contained more than 85% by mass of that substance. Furthermore, the viscosity of a 1% by mass aqueous solution was measured using a B-type viscometer at 25°C and 60 rpm, and was found to be 10 mPa·s. 【0018】 The ratio of galactose to mannose content (galactose:mannose) was measured to be 1:1.8. The dietary fiber content was measured by enzyme-HPLC and found to be 89% by mass. The oligosaccharide content was 10% by mass. Furthermore, when the amino acid analysis of the guar bean protein in this example was performed, the values ​​for (glutamic acid + glutamine + aspartic acid + asparagine) / total amino acids, (glutamic acid + glutamine + aspartic acid + asparagine) / (arginine + lysine + histidine), (cystine + tyrosine) / total amino acids, and (valine + histidine + proline + leucine) / total amino acids were calculated to be 0.56, 22.3, 0.16, and 0.03, respectively. Furthermore, when the ratio of guar gum hydrolysate to guar bean protein was determined, it was found to be 418:1. 【0019】 Example 3 900g of water was mixed with 0.1N hydrochloric acid to adjust the pH to 4. Then, 0.25g of β-mannanase derived from Aspergillus bacteria (manufactured by Novo Nordisk Bioindustry) and 100g of guar gum powder were added and mixed. This mixture was reacted at 50°C to 55°C for 12 hours. After the reaction, the enzyme was inactivated by heating at 90°C for 15 minutes. The reaction solution was filtered and separated by suction filtration, and the clear solution obtained by removing insoluble matter was concentrated under reduced pressure (using a Yamato evaporator). A solid content of 20% by mass was obtained. This was dried using a spray dryer (manufactured by Okawara Chemical Machinery Co., Ltd.) to obtain 70g of guar gum hydrolysate as powder. The average molecular weight of the guar gum hydrolysate was determined in the same manner as in Example 1, and was found to be approximately 1.5 × 10⁻⁶. 4 The result was that the average molecular weight was 1.8 × 10⁶. 3 ~2.0×10 5 It contained more than 86% by mass of that substance. 【0020】 Furthermore, the viscosity of a 1% by mass aqueous solution was measured using a B-type viscometer at 25°C and 60 rpm, and was found to be 9 mPa·s. The ratio of galactose to mannose content (galactose:mannose) was measured to be 1:2.0. The dietary fiber content was measured by enzyme-HPLC and found to be 88% by mass. The oligosaccharide content was 9% by mass. Furthermore, when the amino acid analysis of the guar bean protein in this example was performed, the values ​​for (glutamic acid + glutamine + aspartic acid + asparagine) / total amino acids, (glutamic acid + glutamine + aspartic acid + asparagine) / (arginine + lysine + histidine), (cystine + tyrosine) / total amino acids, and (valine + histidine + proline + leucine) / total amino acids were calculated to be 0.48, 9.6, 0.17, and 0.05, respectively. Furthermore, when the ratio of guar gum hydrolysate to guar bean protein was determined, it was found to be 357:1. 【0021】 <PHGG intake test using lifestyle-related disease model mice> 1. Test Method (1) Experimental animals, test feed, and rearing methods Four-week-old male C57BL / 6NJcl mice (from CREA Japan Co., Ltd.) were used. After a one-week preliminary rearing period on AIN-93G feed, the mice were divided into groups and reared for 12 weeks with ad libitum feeding of the test feed. The control group, a high-fat diet, had approximately 30% lard added to the AIN-93G composition so that 60% of the calories came from lipids. In the PHGG group, 5% of the cellulose (dietary fiber) in the high-fat diet composition was completely replaced with PHGG. The PHGG used was that which was the same as in Example 3. On day 77 of rearing, body fat percentage and muscle weight were measured by computed tomography (CT) scan. On day 84, dissection was performed, and blood was collected and organs were removed. A block of approximately 5 mm square was taken from the liver for microarray analysis, immersed overnight at 4°C in RNAlater (Thermo Fisher Scientific, Inc.), and then stored at -80°C. 【0022】 (2) Measurement of muscle weight On the 77th day of rearing, body composition was measured using a Latheta LCT-200 X-ray CT scanner (Hitachi, Ltd.). Under isoflurane anesthesia, measurements were taken at 2 mm intervals from the second rib from the bottom of the abdominal rib to the caudal tip of the pelvis, and muscle mass was calculated using the software provided with the Latheta scanner. (3) Contract analysis of serum components We commissioned the Nagahama Life Science Laboratory to measure total cholesterol (T-CHO), triglycerides (TG), non-esterified fatty acids (NEFA), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), glucose (GLU), and glycoalbumin (GA). (4) Liver DNA microarray After immersion in RNAlater and storage at -80°C, total RNA was extracted from liver samples. Fragmented and labeled sense strand cDNA was synthesized and subjected to DNA microarray analysis. After normalizing the obtained data, a two-group comparison was performed, and genes with significantly altered expression between the high-fat diet group and the PHGG group were extracted. Furthermore, enrichment analysis was performed to investigate the function of differentially expressed genes. 【0023】 2. Test Results (1) Muscle weight Figure 1 shows the results of muscle weight measurement. Muscle weight was significantly higher (p<0.05) in the PHGG group compared to the high-fat diet group. (2) Serum components Figure 2 shows the results of LDL-C measurement. Regarding LDL cholesterol, the PHGG group showed significantly lower levels (p<0.05) compared to the high-fat diet group. (3) DNA microarray analysis of the liver Table 2 shows the genes that were significantly increased (↑) or decreased (↓) in liver DNA microarray analysis. 【0024】 [Table 2] 【0025】 Compared to the high-fat diet group, the PHGG group showed decreased expression of genes involved in fat accumulation and cholesterol biosynthesis, including Cd36, SREBP1, Cyp51, Fdft1, Hmgscs1, and Nsdh1. Furthermore, enrichment analysis indicated that PHGG contributes to suppressing fat accumulation, inhibiting cholesterol elevation, and improving insulin resistance. PHGG intake significantly increased the expression of PGC1α and AMPK, which regulate mitochondrial biosynthesis and energy metabolism. 【0026】 <PHGG intake test in pigs> 1. Test Method (1) Experimental animals, test feed, and rearing methods The experiment was conducted on market pigs raised on general farms. The control group was fed a standard market pig feed, while the PHGG group was fed the same feed as the control group plus 0.06% PHGG, continuously from weaning until shipment. The PHGG used was that of Example 1. Three animals were randomly selected from each group, for a total of six animals, and slaughtered them on the 160th day of rearing. Five days after slaughter, approximately 30g of skeletal muscle was collected from the shoulder area and subjected to free amino acid analysis. (2) Measurement of free amino acids All compounds except tryptophan were analyzed using an automated amino acid analysis method, while tryptophan was analyzed using high-performance liquid chromatography. 【0027】 2. Test Results Table 3 shows the average free amino acid levels measured in the control group and the PHGG measurement group (n=3). Figure 3 shows the measurement results for leucine, isoleucine, valine, and branched-chain amino acids. 【0028】 [Table 3] 【0029】 Compared to the control group (C group), the PHGG group (P group) had significantly higher levels of free leucine, free isoleucine, free valine, and free branched-chain amino acids in skeletal muscle. Furthermore, they had significantly higher levels of free methionine, free threonine, and free essential amino acids. Branched-chain amino acids are essential amino acids that cannot be produced in the body and are known to be endurance amino acids that function as energy in muscles during exercise. They are also known to promote muscle synthesis, suggesting that PHGG may be beneficial for muscle building in elderly and frail individuals. Thus, according to this embodiment, we have been able to provide a composition that can be easily ingested and promotes muscle mass increase.

Claims

[Claim 1] (A) Average molecular weight is 1.8 × 10 3 ~2.0×10 5 It contains (B) guar gum hydrolysate and (B) guar bean protein, with a viscosity of 50 mPa·s or less for a 1% aqueous solution, measured at 25°C and 60 rpm using a B-type viscometer. The guar gum hydrolysate is obtained by hydrolyzing galactomannan polysaccharide, which is contained in the endosperm of guar and has a galactose-to-mannose content ratio (galactose:mannose) in the range of 1:1.5 to 1:2.1, using microbially derived β-mannanase to reduce its molecular weight, and the dietary fiber content as defined by the enzyme-HPLC method is at least 70% by mass, the oligosaccharide content of the guar gum hydrolysate is 15% by mass or less, and among the amino acids in the contained protein, (glutamic acid + glutamine + aspartic acid + asparagine) ≥ 100 mg / 100 g, (cystine + tyrosine + serine + threonine) ≥ 40 mg / 100 g, total amino acids ≥ 150 mg / 100 g, and the ratio of (A) / (B) is 1000 or less, in a composition for increasing muscle mass. [Claim 2] A food or beverage for increasing muscle mass containing the muscle mass increasing composition described in claim 1. [Claim 3] A pharmaceutical product for increasing muscle mass, comprising the muscle mass increasing composition described in claim 1. [Claim 4] A muscle-increasing feed containing the muscle-increasing composition described in claim 1.

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