JP4931375B2 - 抗筋肉疲労剤及び飲食品 - Google Patents
抗筋肉疲労剤及び飲食品 Download PDFInfo
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- JP4931375B2 JP4931375B2 JP2005183200A JP2005183200A JP4931375B2 JP 4931375 B2 JP4931375 B2 JP 4931375B2 JP 2005183200 A JP2005183200 A JP 2005183200A JP 2005183200 A JP2005183200 A JP 2005183200A JP 4931375 B2 JP4931375 B2 JP 4931375B2
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Description
しかし、ヤマブシタケから分離される複合成分に抗筋肉疲労作用があることは知られていない。
培養の培地としては、特に制限はなく、菌の発育に必要な諸栄養が含まれていればよい。すなわち、グルコース、シュークロース、マルトース、でんぷん等の炭素源;硫安、硝安、硝酸ソーダ、尿素等の窒素源;ジャガイモエキス、ニンジンエキス、麦芽エキス、ペプトン、V−8ジュース、麹エキス、酵母エキス、酵母粉、タマネギエキス、コーンスティープリカー等の天然の複合栄養源;リン酸、塩酸、マグネシウム、カルシウム、カリウム、鉄等の無機塩類等が利用できる。この他に生長に必要な微量元素、ビタミンなどは適宜添加してもよい。
培養は通常好気条件下がよく、例えば振とう培養法あるいは通気撹拌培養法が用いられる。培養温度は20〜37℃、特に30℃前後で培養するのが好ましい。培養pHは3.0〜9.0、好ましくは4.5〜7.0が生育に好適である。培養日数は培養条件によって異なるが菌糸体の生育があればよく、通常は2〜30日間で、最大の菌糸体の生産される日数がよい。通気撹拌培養では、通気量0.1〜10容量%、撹拌速度30〜800r.p.m.の範囲で行なうのが好ましい。
培養終了後、培養液を遠心分離あるいは濾過することにより培養菌糸体が培養濾液から分離採取される。
本発明の複合成分の有効投与量は、抽出物として1,000mg〜3,000mg/日(成人)とするのが好ましい。
食品の形態も特に制限されず、ドリンク剤、顆粒剤、錠剤、カプセル剤、ペースト剤等の形態が挙げられる。また、かまぼこ、ちくわ等の練り製品;パン等の発酵食品;粉乳、発酵乳等の乳製品;バター等の油脂製品;水、果汁、牛乳、清涼飲料等の飲料;菓子等の食品に添加して使用することもできる。
このような食品には、保存料、着色料、甘味料、酸化防止剤、増粘安定剤、乳化剤、調味料、防腐剤等の食品添加物、天然物等を用いることができる。
ヤマブシタケ乾燥子実体の細胞壁破砕粉末2,000gに抽出溶媒として85%濃度のエタノール8Lを加え、室温で撹拌しながら1日放置して溶媒抽出を行なった。エタノール可溶画分を吸引濾過して低分子成分を除去した後、残渣を70℃以下で通風しながら乾燥した。次いで、この乾燥したエタノール抽出残渣500gを、100℃の熱水3Lで3時間撹拌しつつ煮沸し、熱水抽出を行なった。
熱水抽出液を冷却し、1%シュウ酸アンモニウム(100℃)及び5%水酸化ナトリウム水溶液(30℃)で順次抽出し、遠心分離したその上澄液に5倍量の99%濃度のエタノールを加え、多糖類を沈殿させ、更に遠心分離して得た沈殿を流水中で透析後、凍結乾燥粉末として粗多糖類(1)を得た。収量は23gであった。
上記で得られた多糖類(1)及び(2)の理化学的性質を表1に示し、スペクトルデータを表2に示す。
また、多糖類(2)20mgを0.3M NaOD・D2Oに溶解して、JEOL製GSX−400スペクトルメーターによって測定した核磁気共鳴スペクトル(1H−NMRスペクトル分析、13C−NMRスペクトル分析)分析の結果、多糖類(2)はキシロース、グルコース、マンノース等を随伴するβ−(1→6):β−(1→3)−D−グルカン蛋白複合体であることが確認された。
ヤマブシタケ菌糸体を、蔗糖(グルコース)30g、ポリペプトン2.0g、酵母エキス5g、硝酸ソーダ7g、酸性リン酸−ソーダ7g、塩化カリウム0.3g、硫酸マグネシウム0.3gを水1Lに溶解し、pHを5.0〜7.0にした培地に植菌した。前記組成の液体培地200mLずつを1L容の三角フラスコに分注し、23〜25℃、3〜5日間通風培養あるいはロータリーシェカーを用いて200r.p.m.にて振盪培養し、菌糸体とともにやや粘稠性のある培養濾液を得た。
遠心分離した得た菌糸体に7倍量の水を加え、95〜100℃にて3時間熱水抽出を行なった。熱水抽出液を冷却し、5倍量の99%濃度のエタノールを加え、多糖類を沈殿させ、遠心分離によって得た沈殿を流水中で透析後、凍結乾燥粉末して粗多糖類(3)を得た。収量は菌糸体1g当り80mgであった。
次いで、上記の菌糸体を培養して得られた培養濾液を1/5容まで減圧濃縮し、これに等量の99%濃度のエタノールを加え、4℃にて一夜放置し生じた沈殿を遠心分離後、真空凍結乾燥して、粗多糖類(4)の粉末を得た。収量は培養濾液1L当り800mgであった。
上記で得られた多糖類(5)及び(6)の理化学的性質を表3に示し、スペクトルデータを表4に示す。
また、多糖類(5)及び(6)それぞれ各20mgを0.3M NaOD・D2Oに溶解して、JEOL製GSX−400スペクトルメーターによって測定した核磁気共鳴スペクトル(1H−NMRスペクトル分析、13C−NMRスペクトル分析)分析の結果、多糖類(5)はグルコキシラン−蛋白複合体でβ−(1→6):β−(1→3)−D−グルカン構造をもつことが確認された。
多糖類(6)もβ−(1→6):β−(1→3)−D−グルカン構造をもつガラクトキシログルカン−蛋白複合体であることが確認された。
ウレタン麻酔(1.0g/10mL/kg、腹腔内注射)を施した雄ラット(日本クレア製Jcl:SD系250〜300g)の十二指腸内にカニューレを挿入後、腹位に固定した。腓腹筋を剥離し、腱の切断末端に糸をつけ、日本光電製 FD−ピックアップを介して、筋の張力を記録した。電気刺激は、エム・イー・コマーシャル製(ME2100)刺激装置から注射針電極を介して、腓腹筋に直接行なった。記録開始後10分間は0.1Hzで安定させ、安定後4Hzで15分間刺激した。腓腹筋を0.1Hzで刺激した時は、筋収縮力は数時間安定し変化しないが、4Hzで刺激すると収縮力は一過性に増加した後に減少した。次いで、上記実施例1で得た多糖類(1)、(2)及び実施例2で得た多糖類(5)、(6)をそれぞれ300mg/kgずつ十二指腸内に注入し、30分間刺激を休止した後、4Hzで2分間刺激し筋収縮力を記録した。以後1時間毎に、3時間まで筋の収縮力の回復を、最初に4Hzで刺激した時の最大収縮力を100として測定した。
摘出心臓標本を用いた試験では、大動脈と後大動脈にカニューレを入れ、心臓内に栄養液であるリンゲル液(塩化ナトリウム6g、塩化カリウム0.07g、塩化カルシウム0.1g、炭酸水素ナトリウム0.1gを1Lの水に溶解した液)を灌流させる。なお、静脈洞(ペースメーカ)が残っているので自動能は残っている。また、この標本には交感神経節後線維(心臓機能に促進的に働く)の終末部と副交感神経節及び節後線維(心臓機能に抑制的に働く)が含まれているので、これらに働く薬剤と心筋に直接働く薬剤の検定ができる。
4週齢のSD系(雌/雄)ラット30匹を以下の3群(各群雌5例、雄5例)、すなわち対照(非投与)群、上記実施例1で得た多糖類(1)投与(3,000mg/kg)群及び多糖類(1)投与(1,000mg/kg)群に群分けし、1日1回30日間経口投与し、尿及び血液の各成分値に対する影響を試験した。その結果、全て正常範囲にあり、対照群との差は認められなかった。
製造例1
実施例1と同様にして得た子実体の熱水抽出液1kg、砂糖1kg、ハチミツ15g、カラメル5g、アスコルビン酸0.75g、クエン酸0.3g及びレモン系香料0.2gを調合し、健康飲料を製造した。
実施例1と同様にしてエタノール抽出残渣を得、次いで該エタノール抽出残渣を凍結乾燥させ、凍結乾燥物を得た。この凍結乾燥物10g、アスコルビン酸0.5g及びリンゴ搾汁液2kgを調合してリンゴジュースを製造した。
採肉後、水さらしして脱水した魚肉類2kgを予冷し、実施例1で得た多糖類(2)5g、若干量の塩化カルシウム及び第三リン酸ナトリウムを添加した。次いで、魚肉類をすりつぶし、成形後凍結して冷凍すり味を得た。
Claims (2)
- ヤマブシタケのきのこ子実体、その菌株の培養物又はそれらの処理物を抽出処理後、更にクロマト分画処理及び/又は膜分離処理して得られるβ−(1→6):β−(1→3)−D−グルカン蛋白複合体を有効成分とすることを特徴とする抗筋肉疲労剤。
- 筋肉疲労が腓腹筋疲労又は心筋疲労である請求項1記載の抗筋肉疲労剤。
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