JP2005027511A

JP2005027511A - Method for culturing p. yucatensis(murr.)imaz.

Info

Publication number: JP2005027511A
Application number: JP2003193127A
Authority: JP
Inventors: Tetsuo Otake; 哲夫大竹; Hiroshi Tanaka; 弘田中
Original assignee: Naris Cosmetics Co Ltd
Current assignee: Naris Cosmetics Co Ltd
Priority date: 2003-07-07
Filing date: 2003-07-07
Publication date: 2005-02-03
Anticipated expiration: 2023-07-07
Also published as: JP4227474B2

Abstract

<P>PROBLEM TO BE SOLVED: To provide a method for culturing P. yucatensis(Mur.)Imaz. relating to a mushroom-bed culturing method improving bleaching effect, specifically improving bleaching effect of P. yucatensis(Murr.)Imaz. belonging to the genus Phellinus. <P>SOLUTION: The method for culturing P. yucatensis(Murr.)Imaz. comprises culturing P. yucatensis(Murr.)Imaz. belonging to the genus Phellinus in a culture medium where mulberry sawdust, rice bran, wheat starch, corn starch are added to obtain bleaching effect which has hitherto not been obtained by a conventional mushroom-bed culturing method. <P>COPYRIGHT: (C)2005,JPO&NCIPI

Description

【０００１】
【発明の属する技術分野】本発明は、美白効果を向上させるキノコの菌床培養方法に関するものであり、更に詳しくは、キコブタケ（Ｐｈｅｌｌｉｎｕｓ）属のメシマコブ（Ｐ．ｙｕｃａｔｅｎｓｉｓ（Ｍｕｒｒ．）Ｉｍａｚ．）の美白効果を向上させるための菌床培養方法に関する。
【０００２】
【従来の技術及び発明が解決しようとする課題】メシマコブは抗腫瘍作用を持つことから、従来から健康食品または漢方薬として利用されている。しかし、その供給源としての天然の子実体は入手が難しく、また入手した後も、子実体の個体差による有用性の変動や量産化等実用面での問題点が多い。これらの問題点を回避する方法として、メシマコブの菌糸を液体培養し、その培養液及び培養菌糸が入浴剤、頭髪料及び皮膚外用剤に利用されている。（例えば、特許文献１、特許文献２、及び特許文献３参照。）
【０００３】
一方近年になって、メシマコブの栽培方法が検討され、一定の条件下で子実体が得られるようになってきた。これにより、天然の子実体において問題点として上がっていた個体差による有用性の変動が回避され、大量供給源の確保を可能にした。（例えば、特許文献４、及び特許文献５参照。）
【０００４】
担子菌類は、胞子が発芽した後、菌糸が成長し、ある特定条件下で子実体が形成される。一般には、この菌糸から分化した子実体が「キノコ」と呼ばれ、市場に出回っているのは菌糸から子実体への栽培が確立したものである。スーパー等で販売されているマイタケ等が一般に良く知られている。また、担子菌類は、菌糸や子実体自体に含まれる成分や代謝物は、摂取する有機物の種類により、分化の段階によってもその種類や量が変動することが知られており、培養、栽培した場合は、培地組成によって大きく左右される。そして、キノコ愛好家の中では、「天然物のマイタケは栽培物より香りが強く、味も良いため、一度食べたら忘れられない。」と言われている。これは、人工の栽培物が天然に生息・発生した子実体とは成分的にも大きく異なることに起因していると考えられている。
【０００５】
上述したように、菌糸は子実体とは形態的にも成分的にも大きく異なっている。更に本発明で言う菌床は、オガクズ培地に菌糸を接種し、培養後オガクズ培地中に菌糸が蔓延した状態のものを指している。これは一般的に、子実体栽培用として調製されたものであって、菌糸の増殖・増産を目的として行われている液体培養物とは明らかに成分的に異なったものである。
【０００６】
以上のように、液体培養により得られた菌糸と、子実体の構成組織として存在する菌糸、また菌床培養により得られた菌糸では各々含有する成分が大きく異なっていると考えられ、また、同じ菌床培養物においても、菌床に添加する組成物により、培養菌糸の成分は、大きく異なってくると考えられている。また、キコブタケ（Ｐｈｅｌｌｉｎｕｓ）属のメシマコブには、美白効果を有することが知られている。（例えば、特許文献６参照）しかし、従来の液体培養等の方法では、その美白効果が十分ではなく、高い美白効果が得られる培養方法の確立が望まれていた。
【０００７】
【特許文献１】
特開昭６１−１２９１１３号公報
【特許文献２】
特開平０７−３１６０２６号公報
【特許文献３】
特開平０７−３１６０３５号公報
【特許文献４】
特開平１１−２６２３２９号公報
【特許文献５】
特開２０００−２３６７４５号公報
【特許文献６】
特開２００３−７３２２５号公報
【０００８】
【問題を解決する手段】本発明者等は、鋭意研究を重ねた結果、キコブタケ（Ｐｈｅｌｌｉｎｕｓ）属のメシマコブ（Ｐ．ｙｕｃａｔｅｎｓｉｓ（Ｍｕｒｒ．）Ｉｍａｚ．）の菌床培養方法として、桑オガクズにその添加剤として、米ぬか及び小麦デンプン及びトウモロコシデンプンからなる組成分による培養抽出物を用いることで、高い美白効果が得られることを見いだした。また、特定の培養組成条件で培養することにより、美白効果が向上することを見いだし、本発明を完成するに至った。
【０００９】
一般的に菌床に用いられる組成物は、オガクズと、栄養分として、米ぬか（生ぬかまたは脱脂ぬか）、トウモロコシぬか、フスマ等が用いられてきた。しかし、本発明者等は種々の栄養物を検討する過程において、栄養物の多い菌床組成物は、培養された菌床培養物の抽出物の美白効果が弱く、栄養分の種類と量を特定することにより、飛躍的に美白効果が向上することをも見いだし、本発明を完成するに至った。
【００１０】本発明で使用するメシマコブ菌床は、特定の配合量の米ぬか及び小麦デンプン及びトウモロコシデンプンと水を添加した桑オガクズの培地より調製する。この桑オガクズ培地を栽培容器へ詰め込み、殺菌・冷却後、上記天然の子実体から公知の方法により分離した菌糸体を接種し、２０〜３０℃で３〜６ヶ月間培養後、オガクズ培地に菌糸が蔓延したものを用いる。
【００１１】本発明で使用する菌床に用いる米ぬかは、米ぬかであれば特に限定はなく、実施例では、太邦株式会社製の市販品を用いた。また、配合量についても有効量配合されていれば特に限定はないが、好ましくは菌床組成物中０．１〜２０重量％、更に好ましくは３．０〜１２重量％である。
【００１２】本発明で使用する菌床に用いるトウモロコシデンプンは、トウモロコシデンプンであれば特に限定はなく、実施例では、三和澱粉株式会社製の市販品を用いた。また配合量ついても有効量配合されていれば特に限定はないが、好ましくは菌床組成物中０．１〜２０重量％、更に好ましくは１．０〜１０重量％である。
【００１３】本発明で使用する菌床に用いる小麦デンプンは、小麦デンプンであれば特に限定はなく、実施例では、三和澱粉株式会社製の市販品を用いた。また配合量ついても有効量配合されていれば特に限定はないが、好ましくは菌床組成物中０．１〜２０重量％、更に好ましくは１．０〜１０重量％である。
【００１４】本発明に用いる桑オガクズは、クワ属のクワであれば特に限定はないが、ヤマクワ（ＭｏｒｕｓｂｏｍｂｙｃｉｓＫｏｉｄｚ）、ハチジョウグワ（ＭｏｒｕｓｋａｇａｙａｍａｅＫｏｉｄｚ）、ノグワ（ＭｏｒｕｓｔｉｌｉａｅｆｏｌｉａＭａｋｉｎｏ）、シマグワ（Ｍｏｒｕｓａｕｓｔｒａｌｉｓ）等が利用できる。
【００１５】本発明に用いるメシマコブ菌床の抽出物の調製法は、特に限定されないが、例えば種々の適当な有機溶媒を用いて低温下から加温下で抽出することができる。抽出溶媒としては、例えば、水；メチルアルコール、エチルアルコール等の低級１価アルコール；グリセリン、プロピレングリコール、１、３−ブチレングリコール等の液状多価アルコール；アセトン、メチルエチルケトン等のケトン；酢酸エチルなどのアルキルエステル；ベンゼン、ヘキサン等の炭化水素；ジエチルエーテル等のエーテル類；ジクロルメタン、クロロホルム等のハロゲン化アルカン等の１種または２種以上を用いることが出来る。そのうち、水、エチルアルコール、１、３−ブチレングリコールの１種または２種以上の混合溶媒が特に好適である。
【００１６】メシマコブ菌床の抽出は、菌床をそのままあるいは乾燥したものを重量比で１〜１０００倍量、特に１０〜１００倍量の溶媒を用いることが出来る。常温抽出の場合には、０℃以上、特に２０℃〜４０℃で１時間以上、特に３〜７日間行うのが好ましい。また、６０〜１００℃で０．５〜２４時間、加熱抽出しても良い。
【００１７】以上のような条件で得られるメシマコブ菌床の抽出物は、ろ過等の処理をして、溶液のまま用いても良いが、更に必要により、濃縮、粉末化したものを適宜使い分けて用いることが出来る。
【００１８】
【発明の実態】以下、本発明によるキコブタケ（Ｐｈｅｌｌｉｎｕｓ）属のメシマコブ〔Ｐ．ｙｕｃａｔｅｎｓｉｓ（Ｍｕｒｒ．）Ｉｍａｚ．〕菌床の抽出物のチロシナーゼ活性抑制効果、メラニン産生抑制効果、にかかわる実施例について説明するが、本発明は、ここに記載された実施例に限定されるものではない。なお、実施例中の数字は、特に断りのない限り重量％を示す。
【００１９】
＜実施例１＞チロシナーゼ活性抑制効果の測定
皮膚の黒化には色素のメラニンが深く関与していると考えられる。すなわち、メラニンが紫外線などの外的刺激を受けて肌の皮膚組織で生産され、その結果、肌の黒化が促進され、シミ・ソバカス・色黒等の症状が引き起こされると考えられている。よって、肌の美白作用の確認法としメラニンの生成に関与するチロシナーゼ（酵素）の活性化を抑制することに着目し、今回、チロシナーゼ（シグマ社製）によるチロシナーゼ活性抑制試験を行った。以下、メシマコブ菌床の美白効果に関するチロシナーゼ活性抑制試験について説明する。
【００２０】
＜菌床培養の組成例１＞
本発明に係る菌床を表１に示した組成にて作製した。米ぬか、フスマは太邦株式会社のものを使用した。小麦デンプン、トウモロコシデンプンは三和澱粉工業株式会社のものを使用した。酵母エキスはアサヒビール株式会社のスーパーミーストパウダーＡ００１を使用した。
【００２１】
【表１】

【００２２】
＜菌床の作製法＞前記組成物に水を加え均一に混合し、オートクレーブにて滅菌する。滅菌後冷却した組成物に長崎県男女群島女島より採取した天然のメシマコブ子実体から無菌的に切片を切除した切片をＰｏｔａｔｏ−ＤｅｘｔｒｏｓｅＢｒｏｔｈ（Ｄｉｆｃｏ社製）２．４％と寒天１．５％より調製したポテト・デキストロース寒天培地上に置き、２４℃で４週間培養後、菌糸を得た。得られた菌糸を表１に示した各菌床組成物に接種後２４℃で３ヶ月間培養し、培地中に菌糸が蔓延した状態の菌床を得た。
【００２３】
（ａ）試料の調製：メシマコブ菌床抽出物は、培養したメシマコブ菌床を５０％エタノール水溶液で常温にて７２時間抽出した。陽性対照として用いたアルブチンは、市販の試薬を用いてＰＢＳ（−）溶液に溶解した。試料は全て、０．２μｍメンブレンフィルターで除菌ろ過を行った。メシマコブ菌床抽出物の濃度は１．０％濃度に調整した。
（ｂ）測定方法：試験管にそれぞれＬ−チロシン溶液（濃度：０．３ｍｇ／ｍＬ）１ｍＬと、マッキルベイン緩衝液（ＭｃＩｌｖｉｎｅＢｕｆｆｅｒＳｏｌｕｔｉｏｎ）（ｐＨ６．８）１ｍＬとを入れておき、これらの各試験管に前記（ａ）で調製した各供試料の測定用溶液および対照液をそれぞれ０．９ｍＬ加え、これを３７℃の恒温水槽中で１０分間インキュベートする。前記インキュベートしたものにチロシナーゼ溶液（濃度：１ｍｇ／ｍＬ・マッキルベイン緩衝液）を０．１ｍＬ加えて、直ちに４７５ｎｍにおける吸光度を経時的に測定する。（各測定時点での吸光度値として、チロシナーゼ溶液添加直後の吸光度に対しては添字_０を、添加後Ｘ分インキュベート経過後の吸光度値に対しては添字ｘをそれぞれ付して示す）各吸光度値を数１に代入してチロシナーゼ活性抑制率を算出する。なお、この発明におけるのチロシナーゼ活性抑制率の算出には、反応液投入後１０分後の吸光度を使用する。なお、この吸光度値はドーパクロム（メラニンの前駆物質）の生成量により測定されるものである。また、この値が高くなるに従って、チロシナーゼ活性の抑制が高くなっていることを示している。
【００２４】
【数１】
チロシナーゼ活性抑制率（％）＝（（Ｂｘ−Ｂ_０）−（Ａｘ−Ａ_０））×１００／（Ｂｘ−Ｂ_０）
Ｂ_０：対照溶液の添加直後における吸光度値、Ｂ_１０：対照溶液の１０分後における吸光度値
Ａ_０：試験溶液の添加直後における吸光度値、Ａ_１０：試験溶液の１０分後における吸光度値
【００２５】
表１に示した菌床組成物のチロシナーゼ活性抑制効果試験の測定結果を表２に示す。表２からも明確なように、桑オガクズ及び米ぬか及び小麦デンプン及びトウモロコシデンプンをすべて添加した実施例１が、これらの添加物を２種組み合わせた比較例１、比較例２、比較例３と比較して、また、１種組み合わせた比較例４、比較例５、比較例６、桑オガクズだけからなる比較例７とそれぞれ比較して、最も効果が高いことが確認できた。即ち、上記の桑オガクスと添加物３種類をすべて添加した実施例１のチロシナーゼ活性抑制率は６７．８％であり、比較例１〜比較例７の組成物中、最も高いチロシナーゼ活性抑制率を示した。これに対して、桑オガクズと添加物を２種類組み合せた比較例１は、４９．７％、比較例２は、５１．２％、比較例３は、４４．６％であった。また、桑オガクズと添加物を１種組み合わせた比較例４は、４７．３％、比較例５は、４０．６％、比較例６は、４２．３％であった。さらに桑オガクズだけからなる比較例７は、３７．２％であった。以上の結果は、桑オガクズに米ぬか及び小麦デンプン及びトウモロコシデンプンを添加して培養することにより、チロシナーゼ活性抑制率が向上することを示すものである。
【００２６】
【表２】

【００２７】
＜実施例２＞メラニン産生抑制効果の測定
皮膚の黒化には色素のメラニンが深く関与していると考えられる。即ち、メラニンが紫外線などの外的刺激を受けて肌の皮膚組織で生産され、その結果、肌の黒化が促進され、シミ・ソバカス・色黒等の症状が引き起こされると考えられている。よって、肌の美白作用の確認法としメラニン産生抑制効果に着目し、マウスメラノーマ細胞によるメラニン産生抑制試験を行った。以下、メシマコブ菌床の美白効果に関するメラニン産生抑制試験について説明する。
【００２８】
＜菌床培養の組成例２＞
＜菌床の作製法＞組成物に水を加え均一に混合し、オートクレーブにて滅菌する。滅菌後冷却した組成物に長崎県男女群島女島より採取した天然のメシマコブ子実体から無菌的に切片を切除した切片をＰｏｔａｔｏ−ＤｅｘｔｒｏｓｅＢｒｏｔｈ（Ｄｉｆｃｏ社製）２．４％と寒天１．５％より調製したポテト・デキストロース寒天培地上に置き、２４℃で４週間培養後、菌糸を得た。得られた菌糸を表４、表６、表８、表１０、表１２に示した実施例及び比較例組成物に接種後２４℃で３ヶ月間培養し、培地中に菌糸が蔓延した状態の菌床を得た。
【００２９】
＜試料の調製＞
実施例１：メシマコブ菌床抽出物は、メシマコブ菌床を５０％エタノール水溶液で常温にて７２時間抽出した。溶媒を留去し、５％濃度になるように５０％エタノール水溶液に溶解した。陽性対照として用いたアルブチンは、市販の試薬を用いてＰＢＳ（−）溶液に溶解した。試料は全て、０．２μｍメンブレンフィルターで除菌ろ過を行った。
【００３０】
比較例３１（天然子実体の調整）：メシマコブ子実体の天然物は、長崎県男女群島女島より採取した物を用いた。このメシマコブ天然子実体を５０％エタノール水溶液で常温にて７２時間抽出した。その後、溶媒を留去し、５％濃度になるように５０％エタノール水溶液に溶解、調整した。
【００３１】
比較例３２（培養菌糸抽出液の調整）：培養菌糸は、天然の子実体から、上記比較例３１と同方法にて分離した菌糸をチャペック培地にポリペプトン０．２％、酵母エキス０．１％を添加した培地で、２４℃で２週間通気培養し、培養して得られた菌糸を５０％エタノール水溶液で常温にて７２時間抽出した。その後、溶媒を留去し、５％濃度になるように５０％エタノール水溶液に溶解、調整した。
【００３２】
比較例３３（菌糸培養液の調整）：菌糸培養液は、天然の子実体から、上記比較例３１と同方法にて分離した菌糸をチャペック培地にポリペプトン０．２％、酵母エキス０．１％を添加した培地で、２４℃で２週間通気培養し、培養物から得られた菌糸を除去した培養液を用いた。
【００３３】
＜細胞培養＞
培養液は、牛胎児血清５．０％を加えたダルベッコＭＥＭ（Ｄ−ＭＥＭ）培地を用いた。細胞は、マウスメラノーマＢ−１６Ｆ−１０を８ｃｍφのシャーレに植え付けた。植え付け量は４×１０^３／ｃｍ^２とした。植え付けの翌日、実施例２で調製した抽出物試料を乾燥物濃度にして１００ｐｐｍの濃度になるよう添加を行い、添加後３日後に試験を終了した。
【００３４】
＜評価方法＞
直径８ｃｍシャーレに増殖した細胞をセルスクレーパーで集め、透明な遠心チューブに入れ３、０００ｒｐｍ、１０分遠心し細胞を集めた。集まった細胞ペレットを２Ｎ−ＮａＯＨ溶液に溶解し、４０５ｎｍの吸光度を測定した。それぞれ吸光度をコントロールと比較することにより、８段階表示をした。
【００３５】
＜判定基準＞
表３に結果の判定基準を示す。メラニン量はコントロールと比較した吸光度値の割合で示した。なおアルブチンは、メラニン産生抑制の陽性対照物質として示している。
【００３６】
【表３】

【００３７】
本発明に係る菌床を表４に示した組成にて作製し、試料を調製した。
表４に示した菌床組成物及び天然の子実体、培養菌糸抽出液、菌糸培養液を用い場合のメラニン産生抑制効果試験の測定結果を表５に示す。表５より明らかなように、本発明に係わる桑オガクズと米ぬか及び小麦デンプン及びトウモロコシデンプンを添加した実施例１はメラニン産生抑制の判定が１であり、最も高いメラニン産生抑制効果が認められ、天然のメシマコブ子実体を用いた比較例３１は、メラニン産生抑制の判定が２であり、高いメラニン産生抑制効果が認められた。このメラニン産生抑制効果は、メラニン産生抑制効果の高いことで知られているアルブチンの効果（判定３）を上回る高い値である。また、メシマコブ培養菌糸抽出液を用いた比較例３２及びメシマコブ菌糸培養液を用いた比較例３３は、メラニン産生抑制の判定がともに６であり、メラニン産生抑制効果はほとんど認められなかった。同様に桑オガクズの代わりに広葉樹オガクズを添加した比較例８もメラニン産生抑制の判定が８であり、メラニン産生抑制する作用が認められなかった。
【００３８】
この様に、本発明の菌床組成物である桑オガクズと米ぬか及び小麦デンプン及びトウモロコシデンプンを添加した実施例１のメラニン産生抑制効果は、高いメラニン産生抑制効果があるとされているアルブチン及び天然のメシマコブ子実体と比較しても高い効果を示している。そして、桑オガクズを添加して培養した菌床の抽出物には広葉樹を添加して培養した菌床の抽出物には見られない高いメラニン産生抑制効果を認めた。
【００３９】
【表４】

【００４０】
【表５】

【００４１】
発明に係る菌床を表６に示した組成にて作製し、試料を調製した。表６に示した菌床組成物のメラニン産生抑制効果試験の測定結果を表７に示す。表７に示されているように、桑オガクズと米ぬか及び小麦デンプン及びトウモロコシデンプンを全て添加した実施例２は、桑オガクズに添加物を２種組み合わせた比較例１１、比較例１２と、桑オガクズに添加物を１種組み合わせた比較例１５、桑オガクズだけからなる比較例７と比較して、最も効果が高いことが確認できた。すなわち上記桑オガクズと３種類の添加物を全て添加した実施例２は、メラニン産生抑制の判定が１であった。
【００４２】
これに対し、桑オガクズに添加物を２種組み合わせた比較例１１、比較例１２は、メラニン産生抑制の判定が２、比較例９は、メラニン産生抑制の判定が２であった。桑オガクズに添加物を１種組み合わせた比較例１５は、メラニン産生抑制の判定が４であった。桑オガクズだけからなる比較例７は、メラニン産生抑制の判定が５であった。実施例２の桑オガクズに添加した添加物３種の内、小麦デンプンの代わりにフスマを同量添加した比較例１０は、メラニン産生抑制の判定が３であり、小麦デンプン添加のものより低かった。比較例１２の桑オガクズに添加した添加物２種の内、小麦デンプンの代わりにフスマを同量添加した比較例１３は、メラニン産生抑制の判定が４であり、小麦デンプン添加のものより低かった。以上の結果は、フスマを添加して培養することでは、メラニン産生抑制効果が向上しないことを示している。実施例２の桑オガクズに添加した添加物３種の内、小麦デンプンの代わりに酵母エキスを添加した比較例９は、メラニン産生抑制の判定が２であり、小麦デンプン添加のものより低かった。比較例１１の桑オガクズに添加した添加物２種の内、トウモロコシデンプンの代わりに酵母エキスを添加した比較例１４は、メラニン産生抑制の判定が４であり、トウモロコシデンプン添加のものより低かった。比較例１５の桑オガクズに添加した添加物２種の内、米ぬかの代わりに酵母エキスを添加した比較例１６は、メラニン産生抑制の判定が５であり、米ぬか添加のものより低かった。以上の結果は、酵母エキスを添加して培養することでは、メラニン産生抑制効果が向上しないことを示している。以上の結果は、桑オガクズと米ぬか及び小麦デンプン及びトウモロコシデンプンを添加して培養することが、他の添加物を用いて培養したものに比較してメラニン産生抑制効果が明らかに向上することを示している。
【００４３】
【表６】

【００４４】
【表７】

【００４５】
次に、米ぬかの最適添加量を決定する為に、桑オガクズと米ぬかからなる菌床を調製した。即ち、桑オガクズの量は同じで、米ぬかの添加量を変えた表８の組成物を調製した。表８に示した菌床組成物のメラニン産生抑制効果試験の測定結果を表９に示す。表９に示したように、米ぬかが無配合の比較例７は、メラニン産生抑制の判定が５であった。一方、米ぬかの添加量が１％の比較例１７は、メラニン産生抑制の判定が５であり、米ぬか無添加のものと変わらなかった。一方、米ぬか添加量が、３％の比較例１８、６％の比較例１９、１２％の比較例２０のメラニン産生抑制の判定はいずれも４であり、米ぬか無添加の組成物と比較して、メラニン産生抑制効果が高いことがわかった。さらに米ぬか添加量２４％の比較例２１のメラニン産生抑制の判定は５であり、米ぬか無添加のものと同等であった。以上の結果は、米ぬかの添加量は、３〜１２％がメラニン産生抑制効果が高いことを示している。
【００４６】
【表８】

【００４７】
【表９】

【００４８】
次に、小麦デンプンの最適添加量を決定する為に、桑オガクズと米ぬか及び小麦デンプンからなる菌床を調製した。即ち、桑オガクズと米ぬかの量は同じで、小麦デンプンの添加量を変えた表１０の組成物を調製した。表１０に示した菌床組成物のメラニン産生抑制効果試験の測定結果を表１１に示す。表１１に示したように、小麦デンプンが無配合の比較例１５は、メラニン産生抑制の判定が５であった。一方、小麦デンプンの添加量が１％の比較例２２は、メラニン産生抑制の判定が３であり、小麦デンプン無添加の比較例と比較して、メラニン産生抑制効果が高いことがわかった。また、小麦デンプン添加量が、４％の比較例２３、１０％の比較例２４のメラニン産生抑制の判定はいずれも２であり、小麦デンプン無添加の組成物と比較して、メラニン産生抑制効果が高いことがわかった。さらに小麦デンプン添加量１２％の比較例２５のメラニン産生抑制の判定は５であり、小麦デンプン無添加のものと同等であった。以上の結果は、小麦デンプンの添加量は、１〜１０％がメラニン産生抑制効果が高いことを示している。
【００４９】
【表１０】

【００５０】
【表１１】

【００５１】
次に、トウモロコシの最適添加量を決定する為に、桑オガクズと米ぬか及びトウモロコシデンプンからなる菌床を調製した。即ち、桑オガクズと米ぬかの量は同じで、トウモロコシデンプンの添加量を変えた表１２の組成物を調製した。表１２に示した菌床組成物のメラニン産生抑制効果試験の測定結果を表１３に示す。表１３に示したように、トウモロコシデンプンが無配合の比較例１５は、メラニン産生抑制の判定が５であった。一方、トウモロコシデンプンの添加量が１％の比較例２６は、メラニン産生抑制の判定が４であり、２％の比較例２７は、メラニン産生抑制の判定が２であり、４％の組成物２８は、メラニン産生抑制の判定が３であり、１０％の比較例２９は、メラニン産生抑制の判定が４であり、トウモロコシデンプン無添加の組成物と比較して、メラニン産生抑制効果が高いことがわかった。さらにトウモロコシデンプン添加量１２％の比較例３０のメラニン産生抑制の判定は５であり、トウモロコシデンプン無添加のものと同等であった。以上の結果は、トウモロコシデンプンの添加量は、１〜１０％がメラニン産生抑制効果が高いことを示している。
【００５２】
【表１２】

【００５３】
【表１３】

【００５４】
【発明の効果】
桑オガクズと米ぬか及び小麦デンプン及びトウモロコシデンプンからなる組成物にキコブタケ（Ｐｈｅｌｌｉｎｕｓ）属のメシマコブ（Ｐ．ｙｕｃａｔｅｎｓｉｓ（Ｍｕｒｒ．）Ｉｍａｚ．）を培養することにより、天然の子実体及び公知であるアルブチンと比較して、高い美白効果有する培養物を提供することができた。[0001]
BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method for culturing mushroom fungi that improves whitening effect, and more particularly, to P. yucatensis (Murr.) Imaz. The present invention relates to a fungus bed culture method for improving the whitening effect.
[0002]
2. Description of the Related Art Meshima Cobb has been used as a health food or a traditional Chinese medicine since it has an antitumor action. However, it is difficult to obtain a natural fruit body as a supply source, and even after it is obtained, there are many problems in practical use such as fluctuations in usefulness due to individual differences of fruit bodies and mass production. As a method for avoiding these problems, the mycelium of Meshimakobu is liquid-cultured, and the culture solution and the cultured mycelium are used as a bath preparation, a hair preparation, and a skin external preparation. (For example, see Patent Document 1, Patent Document 2, and Patent Document 3.)
[0003]
On the other hand, in recent years, cultivation methods of Meshimakobu have been studied, and fruit bodies have been obtained under certain conditions. As a result, fluctuations in usefulness due to individual differences that have been raised as problems in natural fruiting bodies were avoided, and it became possible to secure a large supply source. (For example, refer to Patent Document 4 and Patent Document 5.)
[0004]
In basidiomycetes, mycelium grows after spores germinate, and fruit bodies are formed under certain specific conditions. In general, fruit bodies differentiated from this mycelium are called “mushrooms”, and what is on the market is established cultivation from mycelia to fruit bodies. Maitakes sold at supermarkets are generally well known. In addition, basidiomycetes are known to vary in type and amount of components and metabolites contained in mycelia and fruit bodies themselves depending on the type of organic matter ingested and depending on the stage of differentiation. The case greatly depends on the medium composition. And among mushroom enthusiasts, it is said that “natural maitake has a stronger fragrance and better taste than cultivated products, so you can't forget it once you eat it.” This is thought to be due to the fact that artificial cultivated materials are significantly different from the naturally occurring fruiting bodies.
[0005]
As described above, the mycelium is greatly different from the fruit body both in form and component. Furthermore, the fungal bed referred to in the present invention refers to a bed in which hyphae are inoculated into a sawdust medium and the hyphae are spread in the sawdust medium after the culture. This is generally prepared for fruit body cultivation and is clearly different from the liquid culture that is used for the purpose of growing and increasing mycelia.
[0006]
As described above, the mycelia obtained by liquid culture, the mycelium present as a constituent tissue of the fruiting body, and the mycelia obtained by the mycelia culture are considered to be greatly different from each other, and the same It is considered that the components of the cultured mycelium vary greatly depending on the composition added to the fungus bed in the fungus bed culture. In addition, it is known that Meshimakobu belonging to the genus Phellinus has a whitening effect. (For example, refer to Patent Document 6) However, in the conventional methods such as liquid culture, the whitening effect is not sufficient, and it has been desired to establish a culture method capable of obtaining a high whitening effect.
[0007]
[Patent Document 1]
Japanese Patent Laid-Open No. 61-129113 [Patent Document 2]
Japanese Patent Laid-Open No. 07-316026 [Patent Document 3]
Japanese Patent Laid-Open No. 07-316035 [Patent Document 4]
JP 11-262329 A [Patent Document 5]
JP 2000-236745 A [Patent Document 6]
Japanese Patent Laid-Open No. 2003-73225
[Means for Solving the Problems] As a result of intensive research, the present inventors have added a method for cultivating P. yucatenisis (Murr.) Imaz. It has been found that a high whitening effect can be obtained by using a rice bran and a culture extract composed of wheat starch and corn starch as an agent. Moreover, it discovered that whitening effect improved by culture | cultivating on specific culture composition conditions, and came to complete this invention.
[0009]
As a composition generally used for a fungus bed, sawdust and rice bran (raw or defatted), corn bran, bran and the like have been used as nutrients. However, in the process of examining various nutrients, the present inventors have identified the type and amount of nutrients because the whitening effect of the extract of the cultured bacterial bed culture is weak in the bacterial bed composition rich in nutrients. As a result, it has been found that the whitening effect is dramatically improved, and the present invention has been completed.
The bed of Meshimakobu used in the present invention is prepared from a rice bran and a mulberry sawdust medium supplemented with wheat starch, corn starch and water. This mulberry sawdust medium is packed in a cultivation container, sterilized and cooled, inoculated with mycelium separated from the natural fruit body by a known method, cultured at 20-30 ° C. for 3-6 months, and then mycelia on the sawdust medium. Use the one that spreads.
The rice bran used in the fungus bed used in the present invention is not particularly limited as long as it is a rice bran. In the examples, commercially available products manufactured by Taiho Co., Ltd. were used. Moreover, there is no particular limitation on the blending amount as long as it is blended in an effective amount, but it is preferably 0.1 to 20% by weight, more preferably 3.0 to 12% by weight in the fungus bed composition.
The corn starch used in the fungal bed used in the present invention is not particularly limited as long as it is corn starch. In the examples, commercial products manufactured by Sanwa Starch Co., Ltd. were used. Further, the amount is not particularly limited as long as it is an effective amount, but preferably 0.1 to 20% by weight, more preferably 1.0 to 10% by weight in the fungus bed composition.
The wheat starch used in the fungus bed used in the present invention is not particularly limited as long as it is wheat starch. In the examples, commercial products manufactured by Sanwa Starch Co., Ltd. were used. Further, the amount is not particularly limited as long as it is an effective amount, but preferably 0.1 to 20% by weight, more preferably 1.0 to 10% by weight in the fungus bed composition.
The mulberry sawdust used in the present invention is not particularly limited as long as it is a mulberry belonging to the genus Mulberry. Etc. are available.
The method for preparing an extract of the bed of Meshimakobu used in the present invention is not particularly limited. For example, the extract can be extracted from a low temperature to a high temperature using various appropriate organic solvents. Examples of the extraction solvent include water; lower monohydric alcohols such as methyl alcohol and ethyl alcohol; liquid polyhydric alcohols such as glycerin, propylene glycol, and 1,3-butylene glycol; ketones such as acetone and methyl ethyl ketone; and ethyl acetate. One or more of alkyl esters; hydrocarbons such as benzene and hexane; ethers such as diethyl ether; and halogenated alkanes such as dichloromethane and chloroform can be used. Of these, water, ethyl alcohol, and one or more mixed solvents of 1,3-butylene glycol are particularly suitable.
[0016] For the extraction of the bed of Meshimakobu, the solvent can be used in an amount of 1 to 1000 times, particularly 10 to 100 times the weight of the dried or dried bed. In the case of normal temperature extraction, it is preferably performed at 0 ° C. or higher, particularly 20 ° C. to 40 ° C. for 1 hour or longer, particularly 3 to 7 days. Moreover, you may heat-extract at 60-100 degreeC for 0.5 to 24 hours.
[0018] The extract of the bed of Messimacob obtained under the above conditions may be used in the form of a solution after filtration or the like, but if necessary, the concentrated and powdered one can be used appropriately. Can be used.
[0018]
The present invention will be described below with reference to the present invention. yucatensis (Murr.) Imaz. Examples relating to the tyrosinase activity inhibitory effect and melanin production inhibitory effect of the fungal bed extract will be described, but the present invention is not limited to the examples described herein. In addition, the number in an Example shows weight% unless there is particular notice.
[0019]
<Example 1> Measurement of Tyrosinase Activity Inhibitory Effect It is considered that the pigment melanin is deeply involved in the darkening of the skin. That is, it is considered that melanin is produced in the skin tissue of the skin by receiving external stimuli such as ultraviolet rays, and as a result, the blackening of the skin is promoted and symptoms such as spots, freckles, and darkness are caused. Therefore, focusing on suppressing the activation of tyrosinase (enzyme) involved in the production of melanin as a method for confirming the skin whitening effect, a tyrosinase activity suppression test using tyrosinase (manufactured by Sigma) was conducted this time. Hereinafter, the tyrosinase activity suppression test relating to the whitening effect of the bed of Messimacob will be described.
[0020]
<Composition example 1 of bacterial bed culture>
The fungus bed according to the present invention was prepared with the composition shown in Table 1. The rice bran and the bran were from Taiko Co., Ltd. Wheat starch and corn starch were from Sanwa Starch Co., Ltd. As the yeast extract, Asahi Breweries Super Mist Powder A001 was used.
[0021]
[Table 1]

[0022]
<Preparation Method of Bacteria Bed> Water is added to the composition and mixed uniformly, and sterilized in an autoclave. A section obtained by aseptically excising a section from a natural fruit body collected from Mejima Island, Nagasaki Prefecture, to a composition cooled after sterilization was Potato-Dextrose Broth (Difco) 2.4% and agar 1.5% The mycelium was obtained after placing on a potato-dextrose agar medium prepared more and culturing at 24 ° C. for 4 weeks. The obtained mycelium was inoculated into each mycelial bed composition shown in Table 1 and cultured at 24 ° C. for 3 months to obtain a mycelium in which the mycelium spread in the medium.
[0023]
(A) Preparation of sample: As for the extract of the bed of Meshimakobu, the cultured bed of Meshimakobu was extracted with a 50% aqueous ethanol solution at room temperature for 72 hours. Arbutin used as a positive control was dissolved in a PBS (−) solution using a commercially available reagent. All samples were subjected to sterilization filtration with a 0.2 μm membrane filter. The concentration of the bed extract of Meshimakobu was adjusted to 1.0%.
(B) Measurement method: 1 mL of L-tyrosine solution (concentration: 0.3 mg / mL) and 1 mL of McIlvine Buffer Solution (pH 6.8) were put in a test tube, respectively. 0.9 mL each of the measurement solution and the control solution for each sample prepared in (a) above are added to the tube, and this is incubated for 10 minutes in a constant temperature water bath at 37 ° C. 0.1 mL of a tyrosinase solution (concentration: 1 mg / mL / Mcquilbain buffer) is added to the incubated product, and the absorbance at 475 nm is immediately measured over time. (As absorbance values at the time of each measurement, subscript ₀ is given for the absorbance immediately after addition of the tyrosinase solution, and subscript x is given for the absorbance value after X minutes of incubation after the addition) Is substituted into Equation 1, and the tyrosinase activity inhibition rate is calculated. In addition, in the calculation of the tyrosinase activity inhibition rate in this invention, the absorbance 10 minutes after the reaction solution is added is used. This absorbance value is measured by the amount of dopachrome (melanin precursor) produced. Moreover, it has shown that suppression of tyrosinase activity has become high as this value becomes high.
[0024]
[Expression 1]
Tyrosinase activity inhibition rate (%) = ((Bx−B ₀ ) − (Ax−A ₀ )) × 100 / (Bx−B ₀ )
B ₀ : Absorbance value immediately after addition of the control solution, B ₁₀ : Absorbance value immediately after the addition of the control solution A ₀ : Absorbance value immediately after the addition of the test solution, A ₁₀ : Absorbance value after 10 minutes of the test solution ]
Table 2 shows the measurement results of the tyrosinase activity inhibitory effect test of the fungus bed composition shown in Table 1. As is clear from Table 2, Example 1 to which all of mulberry sawdust and rice bran and wheat starch and corn starch were added was compared with Comparative Example 1, Comparative Example 2, and Comparative Example 3 in which these additives were combined. In addition, it was confirmed that the effects were the highest in comparison with Comparative Example 4, Comparative Example 5, Comparative Example 6, and Comparative Example 7 consisting only of mulberry sawdust. That is, the tyrosinase activity inhibition rate of Example 1 to which all of the above mulberry ogax and three kinds of additives were added was 67.8%, and the highest tyrosinase activity inhibition rate among the compositions of Comparative Examples 1 to 7 was obtained. Indicated. On the other hand, the comparative example 1 which combined two kinds of mulberry sawdust and additive was 49.7%, the comparative example 2 was 51.2%, and the comparative example 3 was 44.6%. Moreover, the comparative example 4 which combined 1 type of mulberry sawdust and the additive was 47.3%, the comparative example 5 was 40.6%, and the comparative example 6 was 42.3%. Furthermore, the comparative example 7 which consists only of mulberry sawdust was 37.2%. The above results show that tyrosinase activity inhibition rate is improved by adding rice bran, wheat starch and corn starch to mulberry sawdust and culturing.
[0026]
[Table 2]

[0027]
<Example 2> Measurement of melanin production inhibitory effect It is considered that the pigment melanin is deeply involved in the darkening of the skin. That is, it is considered that melanin is produced in the skin tissue of the skin under an external stimulus such as ultraviolet rays, and as a result, the blackening of the skin is promoted to cause symptoms such as spots, freckles, and darkness. Therefore, focusing on the melanin production inhibitory effect as a method for confirming the skin whitening effect, a melanin production inhibition test using mouse melanoma cells was conducted. Hereinafter, the melanin production suppression test regarding the whitening effect of the Meshimakobu fungus bed will be described.
[0028]
<Composition example 2 of bacterial bed culture>
<Method for preparing fungus bed> Add water to the composition, mix uniformly, and sterilize in an autoclave. A section obtained by aseptically excising a section from a natural fruit body collected from Mejima Island, Nagasaki Prefecture, to a composition cooled after sterilization was Potato-Dextrose Broth (Difco) 2.4% and agar 1.5% The mycelium was obtained after placing on a potato-dextrose agar medium prepared more and culturing at 24 ° C. for 4 weeks. The obtained mycelia were cultured for 3 months at 24 ° C. after inoculation into the Examples and Comparative Examples compositions shown in Table 4, Table 6, Table 8, Table 10, and Table 12, and the mycelia were infested in the medium. A fungus bed was obtained.
[0029]
<Preparation of sample>
Example 1: The extract of the fungus bed was extracted with a 50% ethanol aqueous solution at room temperature for 72 hours. The solvent was distilled off and dissolved in a 50% aqueous ethanol solution to a concentration of 5%. Arbutin used as a positive control was dissolved in a PBS (−) solution using a commercially available reagent. All samples were subjected to sterilization filtration with a 0.2 μm membrane filter.
[0030]
Comparative Example 31 (Adjustment of natural fruiting body): As a natural product of Meshimakobu fruiting body, a product collected from Mejima Island, Gender Islands, Nagasaki Prefecture was used. This natural fruit body was extracted with a 50% aqueous ethanol solution at room temperature for 72 hours. Thereafter, the solvent was distilled off, and the mixture was dissolved and adjusted in a 50% aqueous ethanol solution so that the concentration became 5%.
[0031]
Comparative Example 32 (Preparation of Cultured Mycelium Extract): Cultured mycelia were obtained by separating mycelia from natural fruit bodies by the same method as in Comparative Example 31 above in a Chapek medium with 0.2% polypeptone and 0.1% yeast extract. The medium was added to the culture medium and aerated at 24 ° C. for 2 weeks. The mycelium obtained by culturing was extracted with a 50% aqueous ethanol solution at room temperature for 72 hours. Thereafter, the solvent was distilled off, and the mixture was dissolved and adjusted in a 50% aqueous ethanol solution so that the concentration became 5%.
[0032]
Comparative Example 33 (Adjustment of mycelium culture solution): The mycelium culture solution was a mycelium separated from a natural fruiting body by the same method as in Comparative Example 31 above in a Chapek medium with 0.2% polypeptone and 0.1% yeast extract. A culture solution was used, which was cultured with aeration at 24 ° C. for 2 weeks and from which the mycelia obtained from the culture were removed.
[0033]
<Cell culture>
As the culture solution, Dulbecco's MEM (D-MEM) medium supplemented with fetal bovine serum 5.0% was used. For the cells, mouse melanoma B-16 F-10 was planted in an 8 cmφ petri dish. The planting amount was 4 × 10 ³ / cm ² . The day after planting, the extract sample prepared in Example 2 was added to a dry product concentration of 100 ppm, and the test was terminated 3 days after the addition.
[0034]
<Evaluation method>
Cells grown in an 8 cm diameter petri dish were collected with a cell scraper, placed in a transparent centrifuge tube, and centrifuged at 3,000 rpm for 10 minutes to collect the cells. The collected cell pellet was dissolved in 2N-NaOH solution, and the absorbance at 405 nm was measured. Each of the absorbances was compared with the control to display 8 levels.
[0035]
<Criteria>
Table 3 shows the determination criteria for the results. The amount of melanin was shown by the ratio of the absorbance value compared with the control. Arbutin is shown as a positive control substance for inhibiting melanin production.
[0036]
[Table 3]

[0037]
A fungus bed according to the present invention was prepared with the composition shown in Table 4, and a sample was prepared.
Table 5 shows the measurement results of the melanin production inhibitory effect test in the case of using the mycelial bed composition and natural fruit bodies, cultured mycelium extract, and mycelial culture shown in Table 4. As apparent from Table 5, Example 1 to which mulberry sawdust, rice bran and wheat starch and corn starch according to the present invention were added had a determination of suppression of melanin production of 1, and the highest inhibitory effect of melanin production was observed. In Comparative Example 31 using the fruit body of No. 1 melanin production, the suppression of melanin production was 2, and a high melanin production inhibitory effect was observed. This melanin production inhibitory effect is a high value that exceeds the effect of Arbutin (determination 3), which is known to have a high melanin production inhibitory effect. Moreover, in Comparative Example 32 using the Meshimakobu culture mycelium extract and Comparative Example 33 using the Meshimakobu mycelia culture solution, both the determination of melanin production suppression was 6, and the melanin production suppression effect was hardly recognized. Similarly, in Comparative Example 8 in which broad-leaved sawdust was added instead of mulberry sawdust, the determination of suppression of melanin production was 8, and no action of inhibiting melanin production was observed.
[0038]
Thus, the melanin production inhibitory effect of Example 1 to which mulberry sawdust and rice bran, wheat starch, and corn starch, which are the fungus bed composition of the present invention, was added is considered to have a high melanin production inhibitory effect. Compared with the fruiting body of, the effect is high. And the high melanin production inhibitory effect which was not seen in the extract of the fungus bed which added hardwood to the extract of the fungus bed cultured by adding mulberry sawdust was recognized.
[0039]
[Table 4]

[0040]
[Table 5]

[0041]
A fungus bed according to the invention was prepared with the composition shown in Table 6, and a sample was prepared. Table 7 shows the measurement results of the melanin production inhibitory effect test of the fungus bed composition shown in Table 6. As shown in Table 7, Example 2 in which all of mulberry sawdust, rice bran, wheat starch, and corn starch were added was Comparative Example 11, Comparative Example 12, and Mulberry sawdust in which two kinds of additives were combined with mulberry sawdust. Compared with Comparative Example 15 in which one kind of additive was combined and Comparative Example 7 consisting of only mulberry sawdust, it was confirmed that the effect was highest. That is, in Example 2 in which all of the above mulberry sawdust and three kinds of additives were added, the determination of suppression of melanin production was 1.
[0042]
In contrast, Comparative Example 11 and Comparative Example 12 in which two kinds of additives were combined with mulberry sawdust had a determination of suppression of melanin production of 2, and Comparative Example 9 had a determination of suppression of melanin production of 2. In Comparative Example 15 in which one kind of additive was combined with mulberry sawdust, the determination of suppression of melanin production was 4. In Comparative Example 7 consisting only of mulberry sawdust, the determination of suppression of melanin production was 5. Of the three types of additives added to the mulberry sawdust of Example 2, Comparative Example 10 in which the same amount of bran was added instead of wheat starch had a melanin production suppression determination of 3, which was lower than that of wheat starch added. . Of the two additives added to the mulberry sawdust of Comparative Example 12, Comparative Example 13 in which the same amount of bran was added instead of wheat starch had a melanin production inhibition determination of 4, which was lower than that of the wheat starch added. . The above results show that the effect of inhibiting melanin production is not improved by culturing with the addition of bran. Of the three types of additives added to the mulberry sawdust of Example 2, Comparative Example 9 in which yeast extract was added instead of wheat starch had a melanin production inhibition determination of 2, which was lower than that of wheat starch added. Of the two types of additives added to the mulberry sawdust of Comparative Example 11, Comparative Example 14 in which yeast extract was added instead of corn starch had a melanin production suppression determination of 4, which was lower than that of the corn starch added. Of the two types of additives added to the mulberry sawdust of Comparative Example 15, Comparative Example 16 in which yeast extract was added instead of rice bran had a melanin production suppression determination of 5, which was lower than that of rice bran added. The above result has shown that the melanin production inhibitory effect is not improved by adding and cultivating yeast extract. The above results show that mulberry sawdust, rice bran, wheat starch and corn starch are added and cultured, and the melanin production inhibitory effect is clearly improved compared to those cultured using other additives. ing.
[0043]
[Table 6]

[0044]
[Table 7]

[0045]
Next, in order to determine the optimum addition amount of rice bran, a fungus bed composed of mulberry sawdust and rice bran was prepared. That is, the composition of Table 8 was prepared with the same amount of mulberry sawdust and different amounts of rice bran added. Table 9 shows the measurement results of the melanin production inhibitory effect test of the fungus bed composition shown in Table 8. As shown in Table 9, Comparative Example 7 containing no rice bran had a melanin production inhibition determination of 5. On the other hand, in Comparative Example 17 in which the amount of rice bran added was 1%, the determination of suppression of melanin production was 5, which was not different from that of rice bran added. On the other hand, the amount of rice bran added was 3% for Comparative Example 18, 6% for Comparative Example 19 and 12% for Comparative Example 20 for determination of suppression of melanin production was 4 in comparison with the composition without any rice bran added. It was found that the melanin production inhibitory effect was high. Furthermore, the determination of the melanin production suppression of Comparative Example 21 in which the amount of rice bran added was 24% was 5, which was equivalent to that of the rice bran added. The above results indicate that 3-12% of the amount of rice bran added has a high melanin production inhibitory effect.
[0046]
[Table 8]

[0047]
[Table 9]

[0048]
Next, in order to determine the optimum addition amount of wheat starch, a fungal bed comprising mulberry sawdust, rice bran and wheat starch was prepared. That is, the compositions of Table 10 were prepared with the same amounts of mulberry sawdust and rice bran but with different amounts of added wheat starch. Table 11 shows the measurement results of the melanin production inhibitory effect test of the fungus bed composition shown in Table 10. As shown in Table 11, Comparative Example 15 containing no wheat starch had a melanin production inhibition rating of 5. On the other hand, Comparative Example 22 in which the addition amount of wheat starch was 1% had a determination of inhibition of melanin production of 3, and was found to have a higher melanin production inhibitory effect than the comparative example without addition of wheat starch. Moreover, the determination of melanin production suppression of Comparative Example 23 with 4% of wheat starch and Comparative Example 24 of 10% is all 2, and compared with the composition without wheat starch addition, the melanin production inhibitory effect Was found to be expensive. Furthermore, the judgment of the melanin production suppression of the comparative example 25 of wheat starch addition amount 12% was 5, and was equivalent to the thing without wheat starch addition. The above results show that 1 to 10% of the added amount of wheat starch has a high melanin production inhibitory effect.
[0049]
[Table 10]

[0050]
[Table 11]

[0051]
Next, in order to determine the optimum addition amount of corn, a fungal bed comprising mulberry sawdust, rice bran and corn starch was prepared. That is, the compositions of Table 12 were prepared with the same amounts of mulberry sawdust and rice bran but with different amounts of corn starch added. Table 13 shows the measurement results of the melanin production inhibitory effect test of the fungus bed composition shown in Table 12. As shown in Table 13, Comparative Example 15 containing no corn starch had a melanin production inhibition determination of 5. On the other hand, in Comparative Example 26 in which the amount of corn starch added is 1%, the determination of inhibition of melanin production is 4, and in Comparative Example 27 of 2%, the determination of inhibition of melanin production is 2 and 4% of the composition 28 The determination of suppression of melanin production is 3, 10% of Comparative Example 29 has a determination of suppression of melanin production of 4, and the melanin production suppression effect is higher than that of the composition without corn starch added. all right. Furthermore, the determination of the melanin production suppression of the comparative example 30 with a corn starch addition amount of 12% was 5, which was the same as that without corn starch addition. The above results show that 1 to 10% of the added amount of corn starch has a high melanin production inhibitory effect.
[0052]
[Table 12]

[0053]
[Table 13]

[0054]
【The invention's effect】
By cultivating P. yucatenisis (Murr. Imaz.) In a composition comprising mulberry sawdust, rice bran, wheat starch and corn starch, compared with natural fruit bodies and known arbutin Thus, a culture having a high whitening effect could be provided.

Claims

A method for cultivating a fungus bed characterized in that a fungus bed containing mulberry sawdust, rice bran, wheat starch and corn starch is used in a culture method for improving the whitening effect of mushrooms belonging to the genus Phellinus.

The mulberry used for the mulberry sawdust to be mixed in the fungus bed is the mulberry genus (Morus bombycis Koidz) and / or the bee mulberry (Morus kagayamae Koidz) and / or the mulberry (Morus tiliaaefolia morus) The method according to claim 1, wherein 3 to 12% by weight of rice bran is mixed with 1 to 10% by weight of wheat starch and 1 to 10% by weight of corn starch.

The mushroom bed culture method according to claim 1 or 2, wherein the mushroom of the genus Phellinus is P. yucatensis (Murr.) Imaz.