JP2014205621A - セリシンの抽出・精製方法 - Google Patents
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Abstract
【解決手段】(1)カイコ由来の繭糸または繭層を、90℃〜98℃かつ0.05〜0.07Mのアルカリ溶液に浸漬して、セリシンを含む精練液を得る工程、および(2)前記精練液を透析してセリシン溶液を得る工程を含む抽出・精製方法とする。
【選択図】図1
Description
カイコ由来セリシンの抽出、精製の工程を図1に沿って説明する。図1は、本発明のセリシン抽出・精製方法の一実施形態を例示したフローチャートである。
<実施例2>カイコ由来セリシンの分析
(1)SDS−PAGE電気泳動解析
実施例1で得たセリシンの可溶性パウダーを試料として、tricine-SDSポリアクリルアミド電気泳動を行った。具体的には、セリシンの可溶性パウダーを16%(T)、3%(C)の分離ゲル上に濃縮ゲルを重層した垂直型スラブゲルを用いて分離した。陰極槽は、0.2M Tris, 0.1M Tricine, 0.1%(W/V) SDSで、陽極槽は、0.2M Tris-HCl buffer(pH8.9)で、泳動用試料(10μl)は、等量のTricine SDS−PAGE用sample buffer[50mM trice HCl(pH6.8), 4%(w/v)SDS, 12%(w/v)glycerol, 2%(v/v)β-mercaptethanol, 0.01%(w/v)Serva blue G] を加え、3分間熱処理したものを用いた。分子量マーカーには、Proctin Marker Kit(M.W.Range 97,000-14,400 GEヘルスケアバイオサイエンス社)を使用した。泳動は、分離ゲルに入るまでは30V、その後分離ゲルの下端に達するまでは、80Vの低電圧で通電した。泳動後のゲルは、銀染色による染色でタンパク質の検出を行った。銀染色は、Oakleyの方法を改良した銀染色法に従って行った(井上ら1986)。
(2)アミノ酸組成分析
実施例1で得たセリシンの可溶性パウダーを6Nの塩酸1mlに溶解し、減圧封管した。封管したものを加水分解(110℃/24時間)を経て真空乾燥処理した後、Lithium citrate サンプルバッファー(pH2.2)で溶解し、孔径0.45μmのフィルター(直径4mm, Millipore社)で濾過し、分析試料とした。分析試料は、アミノ酸アナライザー(JLC-500/v, 日本電子社)に注入してアミノ酸組成を測定した。
<実施例3>精練時間の検討
精練時間を変更した場合のセリシン溶液への影響について検討した。
<実施例4>吸光度の測定
実施例1で得たセリシンパウダー0.1mgを純水1mlに溶解し、この水溶液を吸光度計(日本分光株式会社)により吸収量を波長スペクトルとして求めた。
Claims (6)
- 以下の工程:
(1)カイコ由来の繭糸または繭層を、90℃〜98℃かつ0.05〜0.07Mのアルカリ溶液に浸漬して、セリシンを含む精練液を得る工程、および
(2)前記精練液を透析してセリシン溶液を得る工程、
を含むことを特徴とするセリシンの抽出・精製方法。 - 精練工程において、繭糸または繭層をアルカリ溶液へ浸漬する時間は、1〜3分間であることを特徴とする請求項1のセリシンの抽出・精製方法。
- 精練工程における繭糸または繭層と、アルカリ溶液との浴比は、1:25〜1:100であることを特徴とする請求項1または2のセリシンの抽出・精製方法。
- 精練工程におけるアルカリ溶液は、NaOHまたはKOHであることを特徴とする請求項1から3のいずれかのセリシンの抽出・精製方法。
- 請求項1から4のいずれかのセリシンの抽出・精製方法によって得られたセリシン。
- 請求項5のセリシンを含む化粧料。
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Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN107484722A (zh) * | 2016-06-12 | 2017-12-19 | 苏州大学 | 分泌的丝素原纤中含有丝胶蛋白质的家蚕品种选育方法 |
WO2021103358A1 (zh) * | 2019-11-25 | 2021-06-03 | 上海科技大学 | 一种连续功能化纤维、装置及其制备方法和用途 |
CN113372426A (zh) * | 2020-03-10 | 2021-09-10 | 森田生医股份有限公司 | 丝胶蛋白萃取物、其制备方法与用途以及包含其之抗氧化组合物 |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH04202435A (ja) * | 1990-11-30 | 1992-07-23 | Seiren Co Ltd | セリシン微粉体およびその製造法 |
JP2001039999A (ja) * | 1999-08-02 | 2001-02-13 | Nisshinbo Ind Inc | セリシンの抽出方法及び粉末状セリシン |
JP2002128691A (ja) * | 2000-10-24 | 2002-05-09 | National Institute Of Agrobiological Sciences | セリシン含有素材、その製造方法およびその使用方法 |
JP2014058464A (ja) * | 2012-09-14 | 2014-04-03 | Iwate Univ | 天蚕セリシンの抽出・精製方法および天蚕セリシンを含む医薬組成物、食品、化粧料 |
-
2013
- 2013-04-10 JP JP2013082512A patent/JP6261109B2/ja active Active
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH04202435A (ja) * | 1990-11-30 | 1992-07-23 | Seiren Co Ltd | セリシン微粉体およびその製造法 |
JP2001039999A (ja) * | 1999-08-02 | 2001-02-13 | Nisshinbo Ind Inc | セリシンの抽出方法及び粉末状セリシン |
JP2002128691A (ja) * | 2000-10-24 | 2002-05-09 | National Institute Of Agrobiological Sciences | セリシン含有素材、その製造方法およびその使用方法 |
JP2014058464A (ja) * | 2012-09-14 | 2014-04-03 | Iwate Univ | 天蚕セリシンの抽出・精製方法および天蚕セリシンを含む医薬組成物、食品、化粧料 |
Non-Patent Citations (1)
Title |
---|
東北蚕糸・昆虫利用研究報告, vol. 第35・36号, JPN6016022189, December 2011 (2011-12-01), pages p.21 * |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN107484722A (zh) * | 2016-06-12 | 2017-12-19 | 苏州大学 | 分泌的丝素原纤中含有丝胶蛋白质的家蚕品种选育方法 |
WO2021103358A1 (zh) * | 2019-11-25 | 2021-06-03 | 上海科技大学 | 一种连续功能化纤维、装置及其制备方法和用途 |
CN113372426A (zh) * | 2020-03-10 | 2021-09-10 | 森田生医股份有限公司 | 丝胶蛋白萃取物、其制备方法与用途以及包含其之抗氧化组合物 |
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