JP2012012327A - Food for myopachynsis promotion, auxiliary food, and supplement - Google Patents
Food for myopachynsis promotion, auxiliary food, and supplement Download PDFInfo
- Publication number
- JP2012012327A JP2012012327A JP2010149911A JP2010149911A JP2012012327A JP 2012012327 A JP2012012327 A JP 2012012327A JP 2010149911 A JP2010149911 A JP 2010149911A JP 2010149911 A JP2010149911 A JP 2010149911A JP 2012012327 A JP2012012327 A JP 2012012327A
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- Prior art keywords
- isoquercitrin
- food
- glucosyl
- muscle
- whey
- Prior art date
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- Granted
Links
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Abstract
Description
本発明は、筋肥大促進のための食品、補助食品、及びサプリメントに関する。 The present invention relates to foods, supplements, and supplements for promoting muscle hypertrophy.
筋肥大のプロセスとして、筋肉に負荷をかけることによって筋繊維にある程度の筋損傷が生じると、筋繊維の回復の際に元のレベル以上に超回復し、筋肥大が生じるとされている。 As a process of muscle hypertrophy, when a certain amount of muscle damage occurs in muscle fibers by applying a load to the muscles, it is said that the muscle fibers are super-recovered to the original level or higher when muscle fibers are recovered.
一方、筋肉疲労回復や筋肉量増加に関するサプリメントとして、BCAA、アミノ酸、プロテイン、αリポ酸、カルニチンなどを含む商品が開発されている。 On the other hand, products containing BCAA, amino acids, proteins, α-lipoic acid, carnitine and the like have been developed as supplements for recovery from muscle fatigue and increase in muscle mass.
本発明は、筋肥大促進効果を有する、筋肥大促進剤、食品組成物、食品、サプリメントなどを提供することを目的とする。 An object of the present invention is to provide a muscle hypertrophy promoter, a food composition, a food, a supplement and the like having a muscle hypertrophy promoting effect.
本発明者らは、正常マウス、及び筋肥大モデルマウスにおいて、酵素処理イソクエルシトリンが筋肥大を促進する効果を有することを見出し、本発明の完成に至った。 The present inventors have found that enzyme-treated isoquercitrin has an effect of promoting muscle hypertrophy in normal mice and muscle hypertrophy model mice, and completed the present invention.
本発明の一実施態様では、筋肥大促進剤がα-グルコシルイソクエルシトリンを有効成分として含有する。α-グルコシルイソクエルシトリンとして、酵素処理イソクエルシトリンを含有してもよい。 In one embodiment of the present invention, the muscle hypertrophy promoter contains α-glucosyl isoquercitrin as an active ingredient. As α-glucosyl isoquercitrin, enzyme-treated isoquercitrin may be contained.
本発明の一実施態様では、食品組成物がα-グルコシルイソクエルシトリン、及び、カゼインプロテイン、クレアチン、グルタミン、ホエイプロテイン(乳清)、大豆プロテイン、BCAA、アルギニン、シトルリン、オルニチンからなる群から選択される少なくとも一つを含有する。α-グルコシルイソクエルシトリンとして、酵素処理イソクエルシトリンを含有してもよい。 In one embodiment of the present invention, the food composition is selected from the group consisting of α-glucosyl isoquercitrin and casein protein, creatine, glutamine, whey protein (whey), soy protein, BCAA, arginine, citrulline, ornithine. Containing at least one of As α-glucosyl isoquercitrin, enzyme-treated isoquercitrin may be contained.
本発明の一実施態様では、食品が、上記いずれかの食品組成物を含有する。 In one embodiment of the present invention, the food contains any of the food compositions described above.
本発明の一実施態様では、サプリメントがα-グルコシルイソクエルシトリン、及び、カゼインプロテイン、クレアチン、グルタミン、ホエイプロテイン(乳清)、大豆プロテイン、BCAA、アルギニン、シトルリン、オルニチンからなる群から選択される少なくとも一つを含有する。α-グルコシルイソクエルシトリンとして、酵素処理イソクエルシトリンを含有してもよい。 In one embodiment of the present invention, the supplement is selected from the group consisting of α-glucosyl isoquercitrin and casein protein, creatine, glutamine, whey protein (whey), soy protein, BCAA, arginine, citrulline, ornithine. Contains at least one. As α-glucosyl isoquercitrin, enzyme-treated isoquercitrin may be contained.
本発明によって、筋肥大促進効果を有する、筋肥大促進剤、食品組成物、食品、サプリメントなどを提供することができるようになった。 According to the present invention, it has become possible to provide a muscle hypertrophy promoter, a food composition, a food, a supplement, and the like having a muscle hypertrophy promoting effect.
以下、上記知見に基づき完成した本発明の実施の形態を、実施例を挙げながら詳細に説明する。 Hereinafter, embodiments of the present invention completed based on the above knowledge will be described in detail with reference to examples.
なお、本発明の目的、特徴、利点、及びそのアイデアは、本明細書の記載により、当業者には明らかであり、本明細書の記載から、当業者であれば、容易に本発明を再現できる。以下に記載された発明の実施の形態及び具体的な実施例等は、本発明の好ましい実施態様を示すものであり、例示又は説明のために示されているのであって、本発明をそれらに限定するものではない。本明細書で開示されている本発明の意図ならびに範囲内で、本明細書の記載に基づき、様々に修飾ができることは、当業者にとって明らかである。 The objects, features, advantages, and ideas of the present invention will be apparent to those skilled in the art from the description of the present specification, and those skilled in the art can easily reproduce the present invention from the description of the present specification. it can. The embodiments and specific examples of the invention described below show preferred embodiments of the present invention, and are shown for illustration or explanation. It is not limited. It will be apparent to those skilled in the art that various modifications can be made based on the description of the present specification within the spirit and scope of the present invention disclosed herein.
本発明に係る筋肥大促進剤、食品組成物、食品、及びサプリメントは、下式で表されるα-グルコシルイソクエルシトリンを含有する。α-グルコシルイソクエルシトリンの含有量は特に限定されないが、固形分換算で0.0006%以上、好ましくは0.003%以上、さらに好ましくは0.03%以上であることが好ましい。
α-グルコシルイソクエルシトリンの製造方法は、特に限定されないが、例えば、特公昭54−32073号公報に記載されているように、イソクエルシトリンと澱粉質などのグルコース供与体を含有する溶液に対して糖転移酵素を作用させることにより、イソクエルシトリンにグルコース供与体からグルコース残基を等モル以上転移させ、次いで、アミラーゼを作用させてα-グルコシルイソクエルシトリンを生成させればよい。この方法によって製造したα-グルコシルイソクエルシトリンは、酵素処理イソクエルシトリン(enzyme-modified isoquercitrin: EMIQ)と呼ばれる。さらに、多孔性合成吸着剤に接触させて、その吸着性の違いを利用することにより、α-グルコシルイソクエルシトリンを精製することも可能である。 The method for producing α-glucosyl isoquercitrin is not particularly limited. For example, as described in JP-B-54-32073, a solution containing isoquercitrin and a glucose donor such as starch can be used. Then, a glycosyltransferase is allowed to act to transfer equimolar or more of a glucose residue from the glucose donor to isoquercitrin, and then amylase is allowed to act to produce α-glucosylisoquercitrin. Α-Glucosyl isoquercitrin produced by this method is called enzyme-modified isoquercitrin (EMIQ). Furthermore, α-glucosyl isoquercitrin can be purified by contacting with a porous synthetic adsorbent and utilizing the difference in adsorbability.
特に、日本国特許第2926411号に記載されているように、反応基質として、0.5w/v%以上、好ましくは約1.0〜20.0W/V%の高濃度懸濁状イソクエルシトリンや、アルカリ側pHで溶解若しくは有機溶媒水溶液で溶解させた0.5w/v%以上、好ましくは約1.0〜20.0W/V%の高濃度溶液状イソクエルシトリンを用いることで、高濃度の酵素処理イソクエルシトリンを得ることができる。 In particular, as described in Japanese Patent No. 2926411, as a reaction substrate, 0.5 w / v% or more, preferably about 1.0 to 20.0 W / V% high-concentration suspended isoquercitrin or alkaline side By using high-concentration solution-like isoquercitrin dissolved at pH or dissolved in an organic solvent aqueous solution at 0.5 w / v% or more, preferably about 1.0 to 20.0 W / V%, a high concentration of enzyme-treated isoquercitrin can be obtained. Obtainable.
このように、EMIQの主成分はα-グルコシルイソクエルシトリンであるが、精製次第で、基質や生成過程で生じる中間体などの夾雑物を含有しうる。なお、イソクエルシトリンはアルカリ性水溶液には可溶であるが、水、酸性水溶液に難溶であるのに対し、EMIQは水溶性であり、従来、酸化防止剤、食品添加物などに用いられてきた。 Thus, although the main component of EMIQ is α-glucosyl isoquercitrin, it may contain impurities such as substrates and intermediates generated in the production process depending on the purification. In addition, isoquercitrin is soluble in alkaline aqueous solution, but hardly soluble in water and acidic aqueous solution, whereas EMIQ is water-soluble and has been conventionally used for antioxidants, food additives and the like. It was.
EMIQの製造に用いられるイソクエルシトリンは、高度に精製されたイソクエルシトリンに限定されず、イソクエルシトリンと、例えば、シトロニン、ナリンジン、ヘスペリジンなどのフラボノイド配糖体との混合物、更には、イソクエルシトリンを含有している各種植物由来の抽出物、またはその部分精製物などを適宜使用できる。ここで用いられる植物組織は、イソクエルシトリンを含有すれば特に限定されないが、例えば、ソバの葉茎、エンジュのつぼみ(槐花または槐米)、エニシダのつぼみ、ユーカリの葉茎、イチョウの葉茎、柑橘類果実などを用いることができる。 Isoquercitrin used for the production of EMIQ is not limited to highly purified isoquercitrin, and is a mixture of isoquercitrin and flavonoid glycosides such as citronin, naringin, hesperidin, Extracts derived from various plants containing ercitrin or partially purified products thereof can be used as appropriate. The plant tissue used here is not particularly limited as long as it contains isoquercitrin. For example, buckwheat leaf stem, Enju bud (crab or glutinous rice), Enishi bud, Eucalyptus leaf stalk, Ginkgo biloba leaf Stems, citrus fruits and the like can be used.
特に、イソクエルシトリンとして、日本国特許第2926411号に記載されているように、高濃度イソクエルシトリンを用いることが好ましい。高濃度イソクエルシトリンは、イソクエルシトリンを高濃度、例えば0.5w/v%以上、好ましくは約1.0〜20.0W/V%に含有するものであればよい。この高濃度イソクエルシトリンは、イソクエルシトリンを懸濁することにより、または、pH7.0を越えるアルカリ性水溶液や、有機溶媒を含有する水溶液にイソクエルシトリンを溶解させることにより作製することができる。 In particular, as described in Japanese Patent No. 2926411, it is preferable to use high concentration isoquercitrin as isoquercitrin. The high concentration isoquercitrin may be one containing isoquercitrin at a high concentration, for example, 0.5 w / v% or more, preferably about 1.0 to 20.0 W / V%. This high-concentration isoquercitrin can be produced by suspending isoquercitrin, or by dissolving isoquercitrin in an alkaline aqueous solution exceeding pH 7.0 or an aqueous solution containing an organic solvent.
アルカリ性水溶液としては、例えばカセイソーダ水、アンモニア水などを用いることができる。また、水溶液に含有させる有機溶媒としては、水と互いに溶解しうる有機溶媒であればよく、例えば、メタノール、エタノール、n−プロパノール、イソプロパノール、n−ブタノール、アセトール、アセトンなどの低級アルコール、低級ケトンなどを用いることができる。水溶液中の有機溶媒濃度は、イソクエルシトリンが高濃度に溶解し、かつ、澱粉質や糖転移酵素が不溶化せず、α−グルコシルイソクエルシトリンが生成する濃度であれば特に限定されないが、通常、約3〜70V/V%、好ましくは、約5〜60V/V%とする。また、イソクエルシトリンをできるだけ高濃度に容易に溶解するためには、予め、イソクエルシトリンを上記のアルカリ性水溶液に溶解し、これを有機溶媒水溶液と混合し、更に中和して糖転移反応に利用してもよい。 As alkaline aqueous solution, caustic soda water, ammonia water, etc. can be used, for example. The organic solvent to be contained in the aqueous solution may be an organic solvent that is soluble with water. For example, lower alcohols such as methanol, ethanol, n-propanol, isopropanol, n-butanol, acetol, and acetone, lower ketones Etc. can be used. The concentration of the organic solvent in the aqueous solution is not particularly limited as long as isoquercitrin is dissolved at a high concentration, and starch and glycosyltransferase are not insolubilized and α-glucosylisoquercitrin is generated. About 3 to 70 V / V%, preferably about 5 to 60 V / V%. In order to easily dissolve isoquercitrin at as high a concentration as possible, isoquercitrin is previously dissolved in the above alkaline aqueous solution, mixed with an organic solvent aqueous solution, and further neutralized for the transglycosylation reaction. May be used.
高濃度イソクエルシトリンと共に糖転移酵素を作用させるグルコース供与体は、その糖転移酵素によってイソクエルシトリンからグルコース残基を等モル以上結合したα−グルコシルイソクエルシトリンを生成することのできるものであれば限定されないが、α−グルコシルイソクエルシトリンの生成を容易にするためには、用いる糖転移酵素に適したグルコース供与体を用いることが好ましい。従って、一般的には、アミロース、デキストリン、シクロデキストリン、マルトオリゴ糖などの澱粉部分加水分解物、液化澱粉、糊化澱粉などを用いることができるが、糖転移酵素として、α−グルコシダーゼ(EC 3.2.1.20)を用いる際には、マルトース、マルトトリオース、マルトテトラオースなどのマルトオリゴ糖、またはDE約10〜70の澱粉部分加水分解物などを用いることが好ましく、シクロマルトデキストリン グルカノトランスフェラーゼ(EC 2.4.1.19)を用いる際には、シクロデキストリンまたはDE1以下の澱粉糊化物からDE約60の澱粉部分加水分解部など用いることが好ましく、α−アミラーゼ(EC 3.2.1.1)を用いる際には、DE1以下の澱粉糊化物からDE約30のデキストリン、澱粉部分加水分解物など用いることが好ましい。なお、糖転移反応時の澱粉質濃度は、イソクエルシトリンに対して約0.5〜50倍の範囲が好適である。 Glucose donors that act on glycosyltransferases together with high-concentration isoquercitrins should be able to produce α-glucosylisoquercitrins with equimolar or more glucose residues bound from isoquercitrins by the glycosyltransferases. Although not limited, in order to facilitate the production of α-glucosyl isoquercitrin, it is preferable to use a glucose donor suitable for the glycosyltransferase used. Accordingly, in general, partial starch hydrolysates such as amylose, dextrin, cyclodextrin, maltooligosaccharide, liquefied starch, gelatinized starch and the like can be used, but α-glucosidase (EC 3.2. 1.20), malto-oligosaccharides such as maltose, maltotriose, maltotetraose, or starch partial hydrolyzate having a DE of about 10 to 70 are preferably used, and cyclomaltodextrin glucanotransferase (EC 2.4) is preferably used. When using .1.19), it is preferable to use a starch partially hydrolyzed portion of DE about 60 from cyclodextrin or starch paste of DE1 or less. When α-amylase (EC 3.2.1.1) is used, DE1 It is preferable to use a dextrin having a DE of about 30 or a partially hydrolyzed starch from the following starch paste. The starch concentration during the transglycosylation reaction is preferably in the range of about 0.5 to 50 times that of isoquercitrin.
これらの糖転移酵素の由来は、特に限定されず、例えば、α−グルコシダーゼは、ブタの肝臓、ソバの種子などの動植物組織、または、ムコール(Mucor)属、ペニシリウム(Penicillium)属などに属するカビ、またはサッカロミセス(Saccharomyces)属などに属する酵母などの微生物を栄養培地で培養し得られる培養物などに由来してもよく、シクロマルトデキストリン グルカノトランスフェラーゼは、バチルス(Bacillus)属、クレブシーラ(Klebsiella)属などに属する細菌培養物に由来してもよく、β−アミラーゼは、バチルス属などに属する細菌、または、アスペルギルス(Aspergillus)属などに属するカビ培養物などに由来してもよい。なお、これらの糖転移酵素は、精製して使用してもよいが、通常は、粗酵素として用いることができる。また、市販の糖転移酵素を用いてもよい。 The origin of these glycosyltransferases is not particularly limited. For example, α-glucosidase is a mold belonging to the genus Mucor, Penicillium, or animal and plant tissues such as pig liver and buckwheat seeds. Alternatively, it may be derived from a culture obtained by culturing a microorganism such as yeast belonging to the genus Saccharomyces in a nutrient medium. It may be derived from a bacterial culture belonging to a genus or the like, and β-amylase may be derived from a bacteria belonging to the genus Bacillus or the like, or a mold culture belonging to the genus Aspergillus or the like. These glycosyltransferases may be used after purification, but can usually be used as crude enzymes. Commercially available glycosyltransferases may also be used.
高濃度の懸濁状イソクエルシトリンとグルコース供与体を含有するイソクエルシトリン高含有液に糖転移酵素を作用させる場合、反応液のpHは約4.5〜6.5とし、反応液の温度は、糖転移酵素の作用しうる限りで、できるだけ高温、具体的には、約70〜90℃に維持することが好ましい。反応が進行し、イソクエルシトリンがα−グルコシルイソクエルシトリンに変換するにつれて懸濁状イソクエルシトリンが徐々に溶解し、同時に、α−グルコシルイソクエルシトリン、α−マルトシル イソクエルシトリン、α−マルトトリオシル イソクエルシトリン、α−マルトテトラオシル イソクエルシトリン、α−マルトペンタオシル イソクエルシトリン、α−マルトヘキサオシル イソクエルシトリンなどのα−グルコシルイソクエルシトリンが容易に高濃度に生成する。 When glycosyltransferase is allowed to act on a high-concentration isoquercitrin-containing solution containing high concentrations of suspended isoquercitrin and glucose donor, the pH of the reaction solution is about 4.5 to 6.5, and the temperature of the reaction solution is sugar transfer It is preferable to maintain the temperature as high as possible, specifically about 70 to 90 ° C., as long as the enzyme can act. As the reaction proceeds and isoquercitrin is converted to α-glucosyl isoquercitrin, the suspended isoquercitrin gradually dissolves, and at the same time, α-glucosyl isoquercitrin, α-maltosyl isoquercitrin, α-maltoline. Α-Glucosyl isoquercitrin such as triosyl isoquercitrin, α-maltotetraosyl isoquercitrin, α-maltopentaosyl isoquercitrin, α-maltohexaosyl isoquercitrin is easily produced in high concentration To do.
また、イソクエルシトリンをpH7.0を越えるアルカリ性水溶液に溶解させた高濃度イソクエルシトリン溶液を用いた場合、pHが約7.5〜10.0の水に約0.5W/V%以上、好ましくは、約1.0〜5.0W/V%のイソクエルシトリンを加熱溶解し、これに適量のグルコース供与体を溶解して得られるイソクエルシトリン高含有液を、糖転移酵素の作用しうる限り、高いpHと高温、具体的には、pHを約7.5〜10.0、温度を約50〜80℃に維持し、これに糖転移酵素を作用させるとα−グルコシルイソクエルシトリンが容易に高濃度に生成する。この際、アルカリ性溶液中のイソクエルシトリンは、分解を起しやすいので、これを防ぐため、できるだけ遮光、嫌気下に維持するのが望ましい。必要ならば、L−アスコルビン酸、エリソルビン酸などの抗酸化剤を共存させてもよい。 In addition, when a high-concentration isoquercitrin solution in which isoquercitrin is dissolved in an alkaline aqueous solution exceeding pH 7.0 is used, the water having a pH of about 7.5 to 10.0 is about 0.5 W / V% or more, preferably about 1.0 A solution containing a high amount of isoquercitrin obtained by dissolving and dissolving an appropriate amount of glucose donor in a solution of ~ 5.0 W / V% isoquercitrin by heating, with a high pH and high temperature as long as glycosyltransferase can act. Specifically, when the pH is maintained at about 7.5 to 10.0 and the temperature is about 50 to 80 ° C., and glycosyltransferase is allowed to act on this, α-glucosyl isoquercitrin is easily produced at a high concentration. At this time, isoquercitrin in the alkaline solution is likely to be decomposed. Therefore, in order to prevent this, it is desirable to keep it as light-shielded and anaerobic as possible. If necessary, an antioxidant such as L-ascorbic acid or erythorbic acid may coexist.
また、イソクエルシトリンを有機溶媒水溶液中で溶解させた高濃度イソクエルシトリン溶液を用いた場合、イソクエルシトリンを、予め、有機溶媒に加熱溶解し、次いで、これを澱粉質水溶液と混合し、これに糖転移酵素を加えたり、または、イソクエルシトリンと澱粉質とを有機溶媒水溶液に加熱して溶解させ、所定温度に冷却し、これに糖転移酵素を加えたりして、反応させる。 In addition, when a high concentration isoquercitrin solution in which isoquercitrin is dissolved in an organic solvent aqueous solution is used, isoquercitrin is heated and dissolved in an organic solvent in advance, and then mixed with a starch aqueous solution. Glycosyltransferase is added thereto, or isoquercitrin and starch are dissolved in an organic solvent aqueous solution by heating, cooled to a predetermined temperature, and glycosyltransferase is added thereto to react.
更に、これらの条件を組み合せ、例えば、約2.0〜20.0W/V%の懸濁状イソクエルシトリンと適量のグルコース供与体を含有するイソクエルシトリン高含有液をpH約7.5〜10.0、温度約50〜80℃に維持し、これに糖転移酵素を作用させてもよく、同様に、α−グルコシルイソクエルシトリンが容易に高濃度に生成する。 Further, by combining these conditions, for example, a high isoquercitrin-containing solution containing about 2.0 to 20.0 W / V% suspended isoquercitrin and an appropriate amount of glucose donor is adjusted to a pH of about 7.5 to 10.0 and a temperature of about 50. Glucosyltransferase may be allowed to act on this at −80 ° C. Similarly, α-glucosyl isoquercitrin is easily produced at a high concentration.
また、イソクエルシトリンとして、例えば約0.1〜1.0規定のカセイソーダ水溶液、カセイカリ水溶液、炭酸ソーダ水溶液、水酸化カルシウム水、アンモニア水などの強アルカリ性水溶液に約5.0〜20.0W/V%の高濃度に溶解させたものを用い、これに塩酸、硫酸などの酸性水溶液を加えて酵素の作用しうるpHに調整するとともにグルコース供与体を加え、直ちに糖転移酵素を作用させてもよく、α−グルコシルイソクエルシトリンを容易に高濃度に生成させることができる。この際、高濃度に溶解させたイソクエルシトリン溶液も、酸性水溶液でpH調整するとイソクエルシトリンが析出し易いので、そのpH調整前に、グルコース供与体や少量のα−グルコシルイソクエルシトリンなどを共存させてイソクエルシトリンの析出を抑制しつつ糖転移反応を開始することが好ましい。 In addition, as isoquercitrin, for example, dissolved in caustic soda aqueous solution, caustic potash aqueous solution, sodium carbonate aqueous solution, calcium hydroxide aqueous solution, aqueous ammonia, etc. with high concentration of about 5.0 to 20.0 W / V%. In this solution, an acidic aqueous solution such as hydrochloric acid or sulfuric acid is added to adjust the pH so that the enzyme can act, and a glucose donor may be added to cause the glycosyltransferase to act immediately. Citrine can be easily produced at a high concentration. At this time, isoquercitrin solution dissolved in a high concentration is also likely to precipitate when pH is adjusted with an acidic aqueous solution. Therefore, before adjusting the pH, a glucose donor or a small amount of α-glucosylisoquercitrin may be added. It is preferable to start the transglycosylation reaction while suppressing the precipitation of isoquercitrin in the coexistence.
以上のようにして得られるα−グルコシルイソクエルシトリン含有溶液は、通常、室温下、中性付近で、大量のα−グルコシルイソクエルシトリンと少量の未反応イソクエルシトリンとを溶解含有しており、その合計量が、イソクエルシトリン換算で約5.0〜20.0 W/V%もの高濃度に達する。 The α-glucosyl isoquercitrin-containing solution obtained as described above usually contains a large amount of α-glucosyl isoquercitrin and a small amount of unreacted isoquercitrin at room temperature and near neutrality. The total amount reaches as high as about 5.0 to 20.0 W / V% in terms of isoquercitrin.
糖転移反応により生成させた比較的高分子のα−グルコシルイソクエルシトリンは、必要により、そのままで、または、多孔性合成吸着樹脂により精製した後、グルコアミラーゼ(EC 3.2.1.3)またはβ−アミラーゼ(EC 3.2.1.2)などのアミラーゼによって部分加水分解し、α−グルコシルイソクエルシトリンのα−D−グルコシル基の数を低減させることができる。例えば、グルコアミラーゼを用いれば、α−マルトシル イソクエルシトリン以上の高分子物を加水分解し、グルコースを生成するとともにα−グルコシル イソクエルシトリンを生成させることができ、また、β−アミラーゼを用いれば、α−マルトトリオシル イソクエルシトリン以上の高分子物を加水分解し、マルトースを生成するとともに主にα−グルコシルイソクエルシトリンとα−マルトシル イソクエルシトリンとの混合物を生成させることができる。 The relatively high molecular weight α-glucosyl isoquercitrin produced by the transglycosylation reaction may be used as it is or after purification with a porous synthetic adsorption resin, followed by glucoamylase (EC 3.2.1.3) or β-amylase. Partial hydrolysis with amylases such as (EC 3.2.1.2) can reduce the number of α-D-glucosyl groups of α-glucosyl isoquercitrin. For example, glucoamylase can be used to hydrolyze α-maltosyl isoquercitrin or higher macromolecules to produce glucose and α-glucosyl isoquercitrin, and β-amylase can be used. , Α-maltotriosyl isoquercitrin or higher polymer can be hydrolyzed to produce maltose and to produce a mixture of α-glucosylisoquercitrin and α-maltosyl isoquercitrin mainly.
以上のようにして得られたα−グルコシルイソクエルシトリンを含有する溶液は、そのままでα−グルコシルイソクエルシトリン製品にすることもできる。さらに、溶液を濾過、濃縮してシロップ状にしたり、更に乾燥、粉末化して粉末状にしたりすることもできる。 The solution containing α-glucosyl isoquercitrin obtained as described above can be directly used as an α-glucosyl isoquercitrin product. Furthermore, the solution can be filtered and concentrated to form a syrup, or further dried and pulverized into a powder.
さらに、精製されたα−グルコシルイソクエルシトリン製品を製造する場合には、多孔性合成吸着剤による吸着性の差を利用し、α−グルコシルイソクエルシトリンとグルコース供与体などの夾雑物を分離すればよい。また、多孔性合成吸着剤による精製は、グルコース供与体、水溶性糖類だけでなく、水溶性の塩類などの夾雑物も同時に除去できる特長を有している。 Furthermore, when producing a purified α-glucosyl isoquercitrin product, use the difference in adsorptivity by the porous synthetic adsorbent to separate α-glucosyl isoquercitrin and impurities such as glucose donors. That's fine. In addition, purification with a porous synthetic adsorbent has the advantage that not only glucose donors and water-soluble saccharides but also impurities such as water-soluble salts can be removed at the same time.
多孔性合成吸着剤は、多孔性で広い吸着表面積を有し、かつ非イオン性のスチレン−ジビニルベンゼン重合体、フェノール−ホルマリン樹脂、アクリレート樹脂、メタアクリレート樹脂などの合成樹脂であり、例えば、市販されているRohm & Haas社製造の商品名アンバーライトXAD−1、アンバーライトXAD−2、アンバーライトXAD−、アンバーライトXAD−7、アンバーライトXAD−8、アンバーライトXAD−11、アンバーライトXAD−12、三菱化成工業株式会社製造の商品名ダイヤイオンHP−10、ダイヤイオンHP−20、ダイヤイオンHP−30、ダイヤイオンHP−40、ダイヤイオンHP−50、IMACTI社製造の商品名イマクティSyn−42、イマクティSyn−44、イマクティSyn−46などが例示できる。 The porous synthetic adsorbent is a synthetic resin such as a non-ionic styrene-divinylbenzene polymer, a phenol-formalin resin, an acrylate resin, and a methacrylate resin, for example, commercially available. Product names manufactured by Rohm & Haas Amberlite XAD-1, Amberlite XAD-2, Amberlite XAD-, Amberlite XAD-7, Amberlite XAD-8, Amberlite XAD-11, Amberlite XAD- 12. Product names manufactured by Mitsubishi Chemical Industries, Ltd. Diaion HP-10, Diaion HP-20, Diaion HP-30, Diaion HP-40, Diaion HP-50, Product names manufactured by IMACTI Syn- Syn- 42, Imakti Syn-44, Imakti Syn-46, and the like.
これらの多孔性合成吸着剤をカラムに充填し、α−グルコシルイソクエルシトリン溶液をカラムに通液すると、α−グルコシルイソクエルシトリンおよび比較的少量の未反応イソクエルシトリンが多孔性合成吸着剤に吸着するのに対し、多量に共存するグルコース供与体、水溶性糖類は吸着されることなくそのまま流出する。 When these porous synthetic adsorbents are packed into a column and an α-glucosyl isoquercitrin solution is passed through the column, α-glucosyl isoquercitrin and a relatively small amount of unreacted isoquercitrin become porous synthetic adsorbents. While adsorbed, glucose donors and water-soluble saccharides coexisting in large quantities flow out without being adsorbed.
特に、α−グルコシルイソクエルシトリン溶液が有機溶媒を含有している場合には、その有機溶媒濃度を除去した後に多孔性合成吸着剤と接触させて精製することが好ましい。 In particular, when the α-glucosyl isoquercitrin solution contains an organic solvent, it is preferable to purify it by contacting with a porous synthetic adsorbent after removing the organic solvent concentration.
必要ならば、糖転移酵素反応終了後、多孔性合成吸着剤に接触させるまでの間に、例えば、反応液を加熱して生じる不溶物を濾過して除去したり、ケイ酸アルミン酸マグネシウム、アルミン酸マグネシウムなどで処理して反応液中の蛋白性物質などを吸着除去したり、強酸性イオン交換樹脂(H型)、中塩基性または弱塩基性イオン交換樹脂(OH型)などで処理して脱塩するなどの精製方法などによって部分精製してもよい。 If necessary, after the glycosyltransferase reaction and before contacting with the porous synthetic adsorbent, for example, the reaction solution is heated to remove insoluble matter, or magnesium aluminate silicate, aluminan. Treat with magnesium acid to adsorb and remove proteinaceous substances in the reaction solution, or treat with strong acidic ion exchange resin (H type), medium basic or weakly basic ion exchange resin (OH type), etc. Partial purification may be performed by a purification method such as desalting.
多孔性合成吸着剤カラムに選択的に吸着したα−グルコシルイソクエルシトリンと比較的少量の未反応イソクエルシトリンは、カラムを希アルカリ、水などで洗浄した後、比較的少量の有機溶媒または有機溶媒と水との混合液、例えば、メタノール水、エタノール水などを通液することにより、まず、α−グルコシルイソクエルシトリンを溶出させ、その後、通液量を増したり有機溶媒濃度を高めたりすることによって未反応イソクエルシトリンを溶出させることができる。 The α-glucosyl isoquercitrin selectively adsorbed on the porous synthetic adsorbent column and a relatively small amount of unreacted isoquercitrin are washed with a dilute alkali, water, etc. By passing a liquid mixture of a solvent and water, for example, methanol water, ethanol water, etc., first, α-glucosyl isoquercitrin is eluted, and then the flow rate is increased or the organic solvent concentration is increased. Thus, unreacted isoquercitrin can be eluted.
このα−グルコシルイソクエルシトリン高含有溶出液に対し、まず有機溶媒を溜去した後、適当な濃度にまで濃縮すればα−グルコシルイソクエルシトリンを主成分とするシラップ状製品が得られる。更に、これを乾燥し粉末化することによって、α−グルコシルイソクエルシトリンを主成分とする粉末状製品が得られる。 If the organic solvent is first distilled off from the eluate containing a high content of α-glucosyl isoquercitrin and then concentrated to an appropriate concentration, a syrup-like product containing α-glucosyl isoquercitrin as a main component can be obtained. Further, by drying and pulverizing this, a powdery product containing α-glucosyl isoquercitrin as a main component can be obtained.
この有機溶媒によるα−グルコシルイソクエルシトリンおよび未反応イソクエルシトリンの溶出操作は、同時に、多孔性合成吸着剤の再生操作にもなるので、この多孔性合成吸着剤の操り返し使用を可能にする。 Since the elution operation of α-glucosyl isoquercitrin and unreacted isoquercitrin with this organic solvent is also a regeneration operation of the porous synthetic adsorbent, the porous synthetic adsorbent can be used repeatedly. .
なお、EMIQは、市販品も入手でき、例えば、サンエミック、サンメリンAO-3000、サンメリンパウダーC-10(三栄源)などを用いてもよい。 In addition, EMIQ can also obtain a commercial item, for example, Sanemik, Sanmerin AO-3000, Sanmerin powder C-10 (Saneigen) etc. may be used.
以上のようにして得られたα−グルコシルイソクエルシトリンを主成分として含んだEMIQ、あるいはα−グルコシルイソクエルシトリンを、常法により剤形化し筋肥大促進剤の有効成分として用いてもよく、食品組成物、食品、及びサプリメントなどの成分として用いてもよい。そして、こうして作製された筋肥大促進剤、食品組成物、食品、及びサプリメントを摂取することによって、筋肥大を促進することができる。特に、筋損傷後、筋繊維の超回復で生じる筋肥大に適用することが好ましい。食品中のEMIQの含有濃度は、適宜決定できるが、固形分換算で0.003W/W%以上であることが好ましく、0.03 W/W%以上であることが好ましい。 EMIQ containing α-glucosyl isoquercitrin as a main component obtained as described above, or α-glucosyl isoquercitrin may be formulated by an ordinary method and used as an active ingredient of a muscle hypertrophy promoter, You may use as ingredients, such as a food composition, a foodstuff, and a supplement. And muscle hypertrophy can be accelerated | stimulated by ingesting the muscle hypertrophy promoter produced in this way, a food composition, a foodstuff, and a supplement. In particular, it is preferably applied to muscle hypertrophy caused by super-recovery of muscle fibers after muscle damage. The content of EMIQ in the food can be determined as appropriate, but is preferably 0.003 W / W% or more, and more preferably 0.03 W / W% or more in terms of solid content.
なお、筋肥大促進剤、食品組成物、食品、及びサプリメントには、α−グルコシルイソクエルシトリンまたはEMIQ以外に、周知の筋強化、筋損傷修復、筋肥大促進、筋疲労回復などの効果を有する物質を共存させても構わない。例えば、カゼインプロテイン、クレアチン、グルタミン、ホエイプロテイン(乳清)、大豆プロテイン、BCAA、アルギニン、シトルリン、オルニチンにα−グルコシルイソクエルシトリンまたはEMIQを加えて、食品組成物としてもよく、それらの少なくとも一つとα−グルコシルイソクエルシトリンまたはEMIQを含んだ食品やサプリメントとしてもよい。なお、これらの物質を含む食品やサプリメントの摂取量や摂取方法は特に限定されないが、物質自体の摂取量や摂取方法に基づいて摂取することが好ましい。 In addition to α-glucosylisoquercitrin or EMIQ, the muscle hypertrophy promoter, food composition, food, and supplement have effects such as well-known muscle strengthening, muscle damage repair, muscle hypertrophy promotion, and muscle fatigue recovery. Substances may coexist. For example, α-glucosyl isoquercitrin or EMIQ may be added to casein protein, creatine, glutamine, whey protein (whey), soy protein, BCAA, arginine, citrulline, ornithine to form a food composition, and at least one of them. It may be a food or supplement containing α-glucosyl isoquercitrin or EMIQ. In addition, although the intake amount and intake method of foods and supplements containing these substances are not particularly limited, it is preferable to take them based on the intake amount or intake method of the substance itself.
本発明において、筋肥大促進効果をもたらすのに有効なEMIQの摂取量は、通常、成人(15歳以上)1日当り1〜500mgであり、好ましくは成人1日当り 5〜350mgである。また体重1kgあたりの摂取量は通常0.015〜8.5mg/kg、より好ましくは0.080〜5.0mg/kgである。 In the present invention, the intake amount of EMIQ effective to bring about the effect of promoting muscle hypertrophy is usually 1 to 500 mg per day for adults (over 15 years old), preferably 5 to 350 mg per day for adults. The intake per kg body weight is usually 0.015-8.5 mg / kg, more preferably 0.080-5.0 mg / kg.
本発明のα−グルコシルイソクエルシトリンまたはEMIQを含む食品組成物は、上記の有効摂取量で、毎日摂取し続けることにより、運動前、運動後の筋肥大促進効果を一層もたらすことができる。 The food composition containing α-glucosyl isoquercitrin or EMIQ of the present invention can further bring about the effect of promoting muscle hypertrophy before and after exercise by continuing to take it daily at the above-mentioned effective intake.
特にホエイプロテインは、非常に吸収がよく、運動後の筋肉の修復、筋肉増強、疲労回復などに効果があるとされている。従って、ホエイプロテインに、筋肥大の促進効果を有するα−グルコシルイソクエルシトリンまたはEMIQを加えることによって、ホエイプロテインの筋肉発育に対する効果が増強される。 In particular, whey protein is very well absorbed and is said to be effective for muscle repair after exercise, muscle strengthening, and recovery from fatigue. Therefore, by adding α-glucosyl isoquercitrin or EMIQ, which has an effect of promoting muscle hypertrophy, to whey protein, the effect of whey protein on muscle growth is enhanced.
本発明の食品組成物や食品の製造に用いるホエイプロテインは、周知の方法によって牛乳や乳製品から製造されるホエイまたはその調製品(濃縮品、乾燥品、粉末品、精製品など)であって、例えば、ホエーパウダー、ホエイプロテインアイソレイトパウダー(WPI)及びホエイプロテインコンセントレイトパウダー(WPC)等の、ホエイプロテインを含む乳製品であればよい。ホエイプロテインを調製するためのホエイには、酸ホエイ、チーズホエイ、膜処理ホエイなどの種類があるが、特に限定されない。例えば、殺菌した牛乳にスターターを加えて発酵させた後、凝固材を加えてカードを収縮させる時に排出させたホエイを、限外濾過およびイオン交換法により濃縮することにより、タンパク質含量90重量%以上の分離ホエイプロテインを製造できる。また、市販のホエイプロテインを用いてもよく、アラセン472(フォンテラ社製)NZMP8899(フォンテラ社製)、WPI 9000(Protient社製)、ホエイプロテイン(森永乳業社製)などを用いてもよい。これらのホエイプロテインに、適宜形状を適合させたα−グルコシルイソクエルシトリンまたはEMIQを混合して、食品組成物、食品、サプリメントとすればよい。 The whey protein used in the production of the food composition or food of the present invention is whey produced from milk or dairy products by a well-known method or a preparation thereof (concentrated product, dried product, powder product, refined product, etc.). For example, it may be a dairy product containing whey protein such as whey powder, whey protein isolate powder (WPI) and whey protein concentrate powder (WPC). There are various types of whey for preparing whey protein, such as acid whey, cheese whey, and membrane-treated whey, but are not particularly limited. For example, after adding a starter to sterilized milk and fermenting it, the whey discharged when the curd is shrunk by adding a coagulant is concentrated by ultrafiltration and ion exchange, so that the protein content is 90% by weight or more. Isolated whey protein. Commercially available whey protein may be used, and Aracene 472 (manufactured by Fontera) NZMP8899 (manufactured by Fontera), WPI 9000 (manufactured by Protient), whey protein (manufactured by Morinaga Milk Industry Co., Ltd.) and the like may be used. These whey proteins may be mixed with α-glucosyl isoquercitrin or EMIQ having an appropriate shape to obtain a food composition, food, or supplement.
なお、本明細書で言及される食品は、健康食品や機能性食品を含み、特定保健用食品や栄養機能食品などの保険機能食品であっても一般食品であってもよい。また、サプリメントは、栄養補給を主な目的とする補助食品のことで、栄養補助食品や健康補助食品や機能性補助食品が含まれ、特定保健用食品や栄養機能食品などの保険機能食品であってもよい。これらは、ヒト用であっても、ヒト以外の哺乳動物用であっても、ヒト以外の脊椎動物用であっても、ヒト以外の動物用であってもかまわない。 The food referred to in this specification includes health foods and functional foods, and may be foods for insurance such as foods for specified health use or foods for nutritional functions, or general foods. Supplements are dietary supplements whose main purpose is nutritional supplements, including dietary supplements, health supplements, and functional supplements, and are insurance functional foods such as specified health foods and functional nutritional foods. May be. These may be for humans, for mammals other than humans, for vertebrates other than humans, or for animals other than humans.
[実施例1]
6週齢のICRマウスを2週間粉末飼料で飼育し、粉末飼料に馴化させた。足底筋に過負荷を与えるため、8週齢時に両脚の腓腹筋とヒラメ筋を切除して切開創を縫合し、その上から接着剤で接着した。対照群(非切除群)については飼育を行わず、その他の群の手術日に足底筋を採取した。各群9−10匹の手術前体重の平均が等しくなるように、均等に割り付けた後、表1に記載の試験食を3週間自由摂取させた。試験食のベースはCE2粉末飼料(日本クレア)とし、そこにEMIQとして0.02%サンエミック(三栄源、15%EMIQ)(EMIQ濃度で、0.003%)を添加した。試験期間中は週3回、体重と摂餌量の測定を行った。(実施例中の%は、全てW/W%を表す)
[Example 1]
Six-week-old ICR mice were bred on a powdered diet for 2 weeks and habituated to the powdered diet. In order to overload the plantar muscles, the gastrocnemius and soleus muscles of both legs were excised at the age of 8 weeks, the incision was sutured, and an adhesive was applied from above. The control group (non-resected group) was not reared, and plantar muscles were collected on the day of surgery for the other groups. After evenly allocating so that the average preoperative weights of 9-10 animals in each group were equal, the test meals listed in Table 1 were freely consumed for 3 weeks. The base of the test meal was CE2 powder feed (CLEA Japan), and 0.02% San-Emic (San-Eigen, 15% EMIQ) (EMIQ concentration, 0.003%) was added thereto as EMIQ. During the test period, body weight and food intake were measured three times a week. (% In the examples all represents W / W%)
手術より3週間後に頸椎脱臼を行い、左脚足底筋の筋腹部分を切り出し、足底筋の凍結切片を作製し、HE染色を行って筋線維断面積および最短直径を測定(NIHイメージ,ver,J1.42)した。データは統計解析用ソフトSPSS(Ver.13.0)を用い、解析を行い、全て平均+/-標準誤差(SE)で示した。足底筋筋線維面積と最短直径について、C群とR群間で、OC群とOR群間の比較をT検定で行った。なお、手術3週間後の総摂餌量、体重および体重増加量、飼料効率について、各群で差はみられなかった。また、手術3週間後の足底筋重量および足底筋重量の体重比も、各群で差が見られなかった。 Three weeks after surgery, cervical dislocation was performed, the abdominal part of the left leg plantar muscle was cut out, frozen section of the plantar muscle was prepared, and HE fiber staining was performed to measure the muscle fiber cross-sectional area and the shortest diameter (NIH image, ver, J1.42). Data was analyzed using the statistical analysis software SPSS (Ver. 13.0), and all data were expressed as mean +/- standard error (SE). Comparison of plantar muscle muscle fiber area and shortest diameter between group C and group R, and between group OC and OR was performed by T test. In addition, there were no differences among the groups regarding the total food intake, body weight and weight gain, and feed efficiency 3 weeks after surgery. In addition, there was no difference in the weight ratio between the plantar muscle weight and the plantar muscle weight 3 weeks after the operation in each group.
図1に示すように、C群よりR群で、またOC群よりOR群で、足底筋筋線維面積と最短直径の両方とも、有意に増加が見られた。即ち、この結果は、0.02%サンエミックの投与により、筋肥大が促進されたことを示している。 As shown in FIG. 1, both the plantar muscle muscle fiber area and the shortest diameter were significantly increased in the R group than in the C group and in the OR group than in the OC group. That is, this result shows that muscle hypertrophy was promoted by administration of 0.02% San-Emic.
[実施例2]
6週齢のICRマウスを、実施例1と同様に処理したが、各群7−8匹を用い、ここでは表2に記載の試験食を自由摂取させた。試験食のベースはAIN93G粉末飼料(日本クレア)のタンパク源をホエイプロテイン(フォンテラ社製、アラセン)に置換したホエイ食とし、そこにEMIQとして0.004%、0.02%、0.2%のサンエミック(三栄源、15%EMIQ)(EMIQ濃度で、0.0006%、0.003%、0.03%)を添加した。試験期間中は週3回、体重と摂餌量の測定を行った。
[Example 2]
Six-week-old ICR mice were treated in the same manner as in Example 1, except that 7 to 8 animals were used in each group, and the test meals listed in Table 2 were freely ingested here. The base of the test meal was a whey meal in which the protein source of AIN93G powder feed (Japan Clare) was replaced with whey protein (Fontera, Aracene), and EMIQ was 0.004%, 0.02%, 0.2% Sun Emic (San-Ei Gen, 15% EMIQ) (0.0006%, 0.003%, 0.03% by EMIQ concentration) was added. During the test period, body weight and food intake were measured three times a week.
実施例1と同様に、筋線維断面積および最短直径を測定し、解析を行った。なお、実施例1と同様、手術3週間後の総摂餌量、体重および体重増加量、飼料効率について、各群で差はみられなかった。また、手術3週間後の足底筋重量および足底筋重量の体重比も、各群で差が見られなかった。 Similar to Example 1, the muscle fiber cross-sectional area and the shortest diameter were measured and analyzed. As in Example 1, there were no differences between the groups in terms of total food intake, body weight and weight gain, and feed efficiency 3 weeks after surgery. In addition, there was no difference in the weight ratio between the plantar muscle weight and the plantar muscle weight 3 weeks after the operation in each group.
図2に示すように、筋線維横断面積については、EMIQを含まないOW群、EMIQを中濃度に含むMR群に対してEMIQを高濃度含むHR群で筋線維横断面積が有意に高値を示す。EMIQを低濃度に含むLR群に対しては、有意水準を5%にとると有意ではないが、p値は0.1であり、EMIQを高濃度含むHR群で、かなり高値を示す傾向が強いといえる。最短直径については、OW群、LR群、MR群のいずれに対しても、HR群は高値を示す傾向が強かった(p=0.159-0.165)。 As shown in FIG. 2, the myofiber cross-sectional area is significantly higher in the myofiber cross-sectional area in the OW group not containing EMIQ and the MR group containing EMIQ at a medium concentration than in the MR group containing EMIQ at a medium concentration. . For the LR group containing EMIQ at a low concentration, although the significance level is not significant at 5%, the p value is 0.1, and the HR group containing a high concentration of EMIQ tends to show a considerably high value. It can be said that it is strong. With regard to the shortest diameter, the HR group tended to show a high value for all of the OW group, the LR group, and the MR group (p = 0.159-0.165).
これらの結果より、ホエイプロテインを餌のベースとした場合、サンエミックの含有量が0.2%以上で効果が確認された。これは、ホエイプロテイン自体に筋損傷の修復効果があるため、EMIQが低濃度では、ホエイプロテインの効果に隠れてしまったのではないかと考えられる。しかしながら、その様な強い効果のある物質であっても、EMIQを適量添加することによって、筋肥大がさらに促進されることが証明された。 From these results, when whey protein was used as the base of the feed, the effect was confirmed when the content of sun emic was 0.2% or more. This is because whey protein itself has an effect of repairing muscle damage, and therefore, it is considered that at a low concentration of EMIQ, it was hidden by the effect of whey protein. However, it has been proved that muscle hypertrophy is further promoted by adding an appropriate amount of EMIQ even for such a substance having a strong effect.
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