TW201828919A - 發酵物及其製造方法 - Google Patents
發酵物及其製造方法 Download PDFInfo
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- TW201828919A TW201828919A TW106146186A TW106146186A TW201828919A TW 201828919 A TW201828919 A TW 201828919A TW 106146186 A TW106146186 A TW 106146186A TW 106146186 A TW106146186 A TW 106146186A TW 201828919 A TW201828919 A TW 201828919A
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- skin
- improvement
- fermented product
- honey
- german chamomile
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Abstract
本發明提供一種發酵物，其係藉由昆氏乳桿菌發酵蜂蜜與德國洋甘菊或其處理物而獲得。
Description
本發明係關於一種發酵物及其製造方法。
已知皮膚、特別是表皮具有障壁功能。所謂障壁功能係防止自生物內過度蒸散水分、或細菌等異物自體外侵入之功能。認為存在於表皮顆粒層之緊密連接(tight junction)於皮膚之障壁功能中擔負重要之作用。緊密連接不僅使鄰接之細胞彼此密接，而且密封細胞與細胞之間隙而控制物質之透過。細胞膜蛋白質之密連蛋白(claudin)、緊連蛋白(occludin)及緊密連接蛋白(tight-junction protein)構成緊密連接。認為該等蛋白質構成緊密連接鏈之骨架，控制緊密連接之障壁功能。於專利文獻1中，記載有一種含有植物之萃取物之密連蛋白產生促進劑及緊連蛋白產生促進劑。於專利文獻2中，記載有一種皮膚障壁功能改善劑，其包含自屬於仙人掌科團扇仙人掌屬之植物之果實獲得之仙人掌果(cactus fruit)果實萃取物。 先前技術文獻 專利文獻 專利文獻1：日本專利5860577號公報 專利文獻2：日本專利特開2012-121871號公報
[發明所欲解決之問題] 於專利文獻1中，記載有使用德國洋甘菊萃取物研究緊連蛋白之產生促進作用。然而，於專利文獻1中，揭示有於德國洋甘菊萃取物中幾乎無緊連蛋白產生促進作用。 本發明之目的在於提供一種具有優異之皮膚改善作用之新穎蜂蜜發酵物。 [解決問題之技術手段] 本發明者等人新發現：使用昆氏乳桿菌發酵蜂蜜與德國洋甘菊或其處理物而獲得之發酵物具有優異之皮膚改善作用。 本發明提供一種發酵物，其係藉由昆氏乳桿菌發酵蜂蜜與德國洋甘菊或其處理物而獲得。該發酵物具有優異之皮膚改善作用。 於上述發酵物中，上述德國洋甘菊或其處理物較佳為德國洋甘菊花之乾燥粉末。藉由使用德國洋甘菊花之乾燥粉末，可更簡便地獲得具有更高之皮膚改善作用之發酵物。 於上述發酵物中，上述蜂蜜與上述德國洋甘菊或其處理物之質量比較佳為26：1～4：1。若質量比為上述範圍，則獲得進而高之皮膚改善作用。 上述發酵物較佳為選自由皮膚障壁功能改善用、皮膚保濕用、皮膚之彈力改善用、皮膚之暗沈改善用、皮膚之皺紋改善用、肌膚粗糙改善用、皮膚之老化防止用、肌膚之肌理改善用及敏感肌膚改善用所組成之群中之至少一種。 本發明亦可理解為用於選自由皮膚障壁功能改善、皮膚保濕、皮膚之彈力改善、皮膚之暗沈改善、皮膚之皺紋改善、肌膚粗糙改善、皮膚之老化防止、肌膚之肌理改善及敏感肌膚改善所組成之群中之至少一種的藉由昆氏乳桿菌發酵蜂蜜與德國洋甘菊或其處理物而獲得之發酵物。 本發明亦可理解為包括將藉由昆氏乳桿菌發酵蜂蜜與德國洋甘菊或其處理物而獲得之發酵物經口投予至需要其之對象之步驟、且應用於該對象之皮膚之步驟的選自由皮膚障壁功能改善方法、皮膚保濕方法、皮膚之彈力改善方法、皮膚之暗沈改善方法、皮膚之皺紋改善方法、肌膚粗糙改善方法、皮膚之老化防止方法、肌膚之肌理改善方法及敏感肌膚改善方法所組成之群中之至少一種。 本發明亦可理解為用於製造選自由皮膚障壁功能改善劑、皮膚保濕劑、皮膚之彈力改善劑、皮膚之暗沈改善劑、皮膚之皺紋改善劑、肌膚粗糙改善劑、皮膚之老化防止劑、肌膚之肌理改善劑及敏感肌膚改善劑所組成之群中之至少一種的藉由昆氏乳桿菌發酵蜂蜜與德國洋甘菊或其處理物而獲得之發酵物之用途。 本發明提供一種食品、化妝品、醫藥品或準藥品，其包含上述發酵物。 又，本發明提供一種發酵物之製造方法，其包括藉由昆氏乳桿菌發酵蜂蜜與德國洋甘菊或其處理物而獲得發酵物之步驟。藉由該製造方法獲得之發酵物具有優異之皮膚改善作用。 於上述製造方法中，德國洋甘菊或其處理物較佳為德國洋甘菊花之乾燥粉末。 於上述製造方法中，上述蜂蜜與上述德國洋甘菊或其處理物之質量比較佳為26：1～4：1。 藉由上述製造方法獲得之發酵物較佳為選自由皮膚障壁功能改善用、皮膚保濕用、皮膚之彈力改善用、皮膚之暗沈改善用、皮膚之皺紋改善用、肌膚粗糙改善用、皮膚之老化防止用、肌膚之肌理改善用及敏感肌膚改善用所組成之群中之至少一種。 又，本發明提供一種食品、化妝品、醫藥品或準藥品之製造方法，其包括添加藉由上述製造方法而獲得之發酵物之步驟。 [發明之效果] 本發明之發酵物具有優異之皮膚改善作用。
以下，詳細地對用以實施本發明之形態進行說明。然而，本發明並不限定於以下之實施形態。於本說明書中，「%」只要未特別提及，則表示「質量%」。 本實施形態之發酵物係藉由昆氏乳桿菌(Lactobacillus kunkeei)發酵蜂蜜與德國洋甘菊或其處理物(以下，亦稱為「德國洋甘菊原料」)而獲得者。 蜂蜜係蜜蜂以自植物之花蜜、樹液、寄生於植物之蟲之分泌液等收集之蜜為主原料製作者。於本實施形態中，供發酵之原料之蜂蜜之種類並無特別限制，例如可使用麥盧卡蜂蜜、洋槐蜂蜜、甘露蜂蜜、苜蓿蜂蜜、橙花蜂蜜、荷花蜂蜜、迷迭香蜂蜜、葵花蜂蜜、油菜花蜂蜜、咖啡花蜂蜜等。蜂蜜可單獨使用1種，亦可併用2種以上。用於採集蜂蜜之蜜蜂之種類、及蜂蜜之產地並無特別限定。 德國洋甘菊係菊科(Compositae)之植物，學名為Matricaria chamomilla L.。作為使用之德國洋甘菊之部位，並無特別限制，可適當地選擇，例如可列舉葉、莖、根、花、蕾、種子、果實、果皮、果核、地上部、全草、該等之混合物等。使用之德國洋甘菊之部位較佳為花。 供發酵之德國洋甘菊原料可為採集德國洋甘菊而獲得之德國洋甘菊其本身，亦可為德國洋甘菊之處理物。作為德國洋甘菊處理物，例如可列舉對德國洋甘菊進行乾燥、粉碎、萃取等處理而獲得之處理物。該等處理可單獨進行1種，亦可併用2種以上。 德國洋甘菊處理物例如可為德國洋甘菊之乾燥物、粉碎物、破碎物、粉末、萃取物等。德國洋甘菊之乾燥例如可藉由太陽光乾燥、熱風乾燥、冷凍乾燥、噴霧乾燥等而進行。粉碎及破碎可分別使用例如粉碎機、破碎機等進行。德國洋甘菊原料較佳為德國洋甘菊之乾燥粉末。德國洋甘菊之乾燥粉末例如可藉由對德國洋甘菊進行乾燥，進而進行粉碎而獲得。德國洋甘菊較佳為於採集後立即進行乾燥、粉碎所得者。作為德國洋甘菊處理物，可使用市售品。 德國洋甘菊之萃取物可藉由通常用於萃取植物之方法自德國洋甘菊萃取而獲得。例如，可藉由如下方式獲得萃取液：於裝滿萃取溶劑之處理槽中投入作為萃取原料之德國洋甘菊之萃取部位，一面視需要適當地進行攪拌，一面溶出可溶性成分，之後進行過濾而去除萃取殘渣。 作為德國洋甘菊之萃取物，可直接使用自德國洋甘菊萃取而獲得之萃取液，亦可使用進而進行例如稀釋、濃縮、乾燥、過濾、純化等處理所得者。 作為萃取溶劑，例如可為水系溶劑、親水性有機溶劑等。作為水系溶劑，例如可列舉純水、純化水、自來水、井水、礦泉水、礦水、溫泉水、泉水、淡水、熱水、離子交換水、生理鹽水、磷酸緩衝液、磷酸緩衝生理鹽水等。作為親水性有機溶劑，例如可列舉甲醇、乙醇、丙醇、異丙醇等碳數為1～5之低級醇；丙酮、甲基乙基酮等低級脂肪族酮；1,3-丁二醇、丙二醇、甘油等碳數為2～5之多元醇等。萃取溶劑可單獨使用該等之1種，亦可併用2種以上。 發酵可使用昆氏乳桿菌進行。用於獲得本實施形態之發酵物之昆氏乳桿菌較佳為表現出對葡萄糖及果糖之合成代謝性者，較佳為表現出對葡萄糖、果糖、蔗糖、海藻糖及葡萄糖酸鹽之合成代謝性者。昆氏乳桿菌亦可為昆氏乳桿菌BPS402。昆氏乳桿菌BPS402係於2011年10月3日以寄存編號FERM P-22177寄存於獨立行政法人產業技術綜合研究所特許生物寄託中心(日本茨城縣筑波市東1丁目1番地1中央第6(郵遞區號305-8566))且可獲取。又，該菌株目前移交於國際寄存，寄存編號為FERM BP-11439。 用於發酵之昆氏乳桿菌之培養可根據常法而進行。培養基只要為可培養該菌者，則無特別限制，可使用天然培養基、合成培養基、半合成培養基等。作為培養基，可使用含有氮源及碳源者，作為氮源，可列舉肉汁、蛋白腖、酪蛋白、酵母萃取液、麩質、大豆粉、大豆水解物、胺基酸等，作為碳源，可列舉葡萄糖、乳糖、果糖、肌醇、山梨糖醇、飴糖、澱粉、麴汁、麥麩、甘蔗渣、糖蜜等。除此之外，可添加無機質(例如，硫酸銨、磷酸鉀、氯化鎂、食鹽、碳酸鈣、鐵、錳、鉬)、各種維生素類等。 培養溫度例如可為4～45℃，較佳為25～40℃，更佳為28～33℃。培養時間例如可為8～72小時。亦可進行通氣振盪或通氣攪拌而進行培養。培養基之pH值例如可為4.0～9.0，較佳為6.0～8.0。作為培養方法，例如可列舉將菌體植菌至MRS培養基，於30℃下培養48小時。 培養昆氏乳桿菌而獲得之菌液可直接接種至包含蜂蜜及德國洋甘菊原料之發酵培養基而用於發酵，亦可藉由培養基等稀釋而使用。 利用昆氏乳桿菌進行之蜂蜜及德國洋甘菊原料之發酵例如可於4～45℃下進行，較佳為於25～40℃下進行。發酵時間例如可為12～60小時，較佳為24～48小時。發酵開始時之含蜂蜜物質之pH值例如可為3～8，較佳為4～6。亦可進行通氣振盪或通氣攪拌而進行發酵。 蜂蜜及德國洋甘菊原料之發酵可藉由在包含蜂蜜及德國洋甘菊原料之發酵培養基中培養昆氏乳桿菌而進行。包含蜂蜜及德國洋甘菊原料之發酵培養基只要為昆氏乳桿菌可發酵之形態即可，例如可為液體、糊劑、凝膠等形態。發酵培養基例如可為於水等溶劑中溶解或懸浮有蜂蜜及德國洋甘菊原料者。 發酵開始時之發酵培養基中之蜂蜜濃度相對於發酵培養基總量例如可為0.1～40%，且可為1～30%，亦可為8～13%。又，發酵開始時之發酵培養基中之蜂蜜濃度以固形物成分計相對於發酵培養基總量例如可為0.08～32%，可為0.8～24%，亦可為6.4～10.4%。 發酵開始時之發酵培養基中之德國洋甘菊原料濃度以固形物成分計相對於發酵培養基總量例如可為0.01～20%，可為0.1～5%，亦可為0.5～2%。發酵開始時之發酵培養基中之蜂蜜(總量)與德國洋甘菊原料(乾燥質量)之質量比例如可為4000：1～1：200，可為26：1～4：1。發酵培養基可進而包含例如維生素類、胺基酸類、礦物質類、鹽類、界面活性劑、脂肪酸、金屬類等在培養乳酸菌時必需之公知之營養素。用於發酵之蜂蜜亦可藉由加熱等而經殺菌處理。 本實施形態之發酵物可為藉由發酵而獲得者本身，亦可為實施過加熱、過濾、純化、濃縮、蒸乾、冷凍乾燥、噴霧乾燥等處理者。本實施形態之發酵物可包含昆氏乳桿菌之活菌體及/或死菌體，亦可藉由過濾等分離去除菌體。本實施形態之發酵物於發酵培養基中包含蜂蜜，故而可期待維生素、胺基酸等成分相較於原料之蜂蜜增加，且產生原料之蜂蜜所不含之有機酸。 本實施形態之發酵物具有選自由緊連蛋白、密連蛋白-1、密連蛋白-4、緊密連接蛋白-1、緊密連接蛋白-2、緊密連接蛋白-3、III型膠原蛋白α1鏈、IV型膠原蛋白、水孔蛋白(aquaporin)3、絲胺酸棕櫚醯轉移酶2及TIMP(tissue inhibitor of metalloproteinase，組織金屬蛋白酶抑制物質)2所組成之群中之至少1種蛋白質之產生促進作用。因此，本實施形態之發酵物亦較佳作為緊連蛋白產生促進用、密連蛋白-1產生促進用、密連蛋白-4產生促進用、緊密連接蛋白-1產生促進用、緊密連接蛋白-2產生促進用、緊密連接蛋白-3產生促進用、III型膠原蛋白α1鏈產生促進用、IV型膠原蛋白產生促進用、水孔蛋白3產生促進用、絲胺酸棕櫚醯轉移酶2產生促進用及/或TIMP2產生促進用。又，上述發酵物亦較佳作為編碼該等蛋白質之各基因之表現促進用。 本實施形態之發酵物具有選自由介白素-1α、介白素-1β、環加氧酶2及基質金屬蛋白酶-9所組成之群中之至少1種蛋白質之產生抑制作用。因此，本實施形態之發酵物較佳作為介白素-1α產生抑制用、介白素-1β產生抑制用、環加氧酶2產生抑制用及/或基質金屬蛋白酶-9產生抑制用。又，上述發酵物亦較佳作為編碼該等蛋白質之各基因之表現抑制用。 關於上述各種蛋白質，已知以下。緊連蛋白、密連蛋白-1、密連蛋白-4、緊密連接蛋白-1、緊密連接蛋白-2及緊密連接蛋白-3係構成緊密連接之蛋白質，參與障壁功能。若緊密連接變脆弱，則引起水分蒸散亢進、保濕成分及鈣離子之流出、與角質層水分量有關之絲聚蛋白及細胞間脂質之合成不良、角質肥厚等，導致肌膚乾燥、肌膚敏感、肌膚粗糙、肌膚無透明感、肌膚之暗沈等。可藉由強化緊密連接而期待預防肌膚乾燥、肌膚敏感、肌膚粗糙、及暗沈、提高透明感等效果。 III型膠原蛋白大量包含於胎兒之皮膚，擔負保持肌膚之彈性之作用。IV型膠原蛋白係存在於表皮與真皮之間之基底膜之主要構成成分。絲胺酸棕櫚醯轉移酶2係合成腦醯胺時之出發速率限制酶。TIMP2係MMP(matrix metalloproteinases，基質金屬蛋白酶)之抑制劑。 已知介白素-1α及介白素-1β促進免疫細胞之活化，引起炎症反應。特別是介白素-1α大量地存在於角化細胞內。已知環加氧酶2係參與炎症之前列腺素之合成酶，因紫外線等而表現上升。環加氧酶2之產生之下降抑制作用於黑色素細胞之前列腺素之產生，抑制因紫外線形成之黃褐斑之產生。 報告有MMP-9分解構成基底膜之蛋白質，因紫外線而表現上升。藉由防止MMP-9之增加，基底膜之破壞得到抑制，鬆弛及皺紋之形成得到抑制。又，藉由防止基底膜之破壞而防止生成於表皮之黑色素向真皮側轉移，藉此抑制難以消除之黃褐斑之產生。 因此，本實施形態之發酵物之皮膚改善作用優異，具體而言，例如皮膚障壁功能改善作用、皮膚保濕作用、皮膚之彈力改善作用、皮膚之暗沈改善作用、皮膚之皺紋改善作用、肌膚粗糙改善作用、皮膚之老化防止作用、肌膚之肌理改善作用及敏感肌膚改善作用優異。因此，本實施形態之發酵物例如較佳作為選自由皮膚障壁功能改善用、皮膚保濕用、皮膚之彈力改善用、皮膚之暗沈改善用、皮膚之皺紋改善用、肌膚粗糙改善用、皮膚之老化防止用、肌膚之肌理改善用及敏感肌膚改善用所組成之群中之至少一種。所謂皮膚之老化防止例如包含預防或改善皮膚之皺紋、黃褐斑、暗沈、鬆弛、粗糙等、改善肌理等。 本發明之一實施態樣係用於選自由皮膚障壁功能改善、皮膚保濕、皮膚之彈力改善、皮膚之暗沈改善、皮膚之皺紋改善、肌膚粗糙改善、皮膚之老化防止、肌膚之肌理改善及敏感肌膚改善所組成之群中之至少一種的藉由昆氏乳桿菌發酵蜂蜜與德國洋甘菊或其處理物而獲得之發酵物。 本發明之一實施態樣係包括將藉由昆氏乳桿菌發酵蜂蜜與德國洋甘菊或其處理物而獲得之發酵物經口投予至需要其之對象之步驟、且應用於該對象之皮膚之步驟的選自由皮膚障壁功能改善方法、皮膚保濕方法、皮膚之彈力改善方法、皮膚之暗沈改善方法、皮膚之皺紋改善方法、肌膚粗糙改善方法、皮膚之老化防止方法、肌膚之肌理改善方法及敏感肌膚改善方法所組成之群中之至少一種。 本發明之一實施態樣係用於製造選自由皮膚障壁功能改善劑、皮膚保濕劑、皮膚之彈力改善劑、皮膚之暗沈改善劑、皮膚之皺紋改善劑、肌膚粗糙改善劑、皮膚之老化防止劑、肌膚之肌理改善劑及敏感肌膚改善劑所組成之群中之至少一種的藉由昆氏乳桿菌發酵蜂蜜與德國洋甘菊或其處理物而獲得之發酵物之用途。 本實施形態之發酵物可直接用作化妝品、醫藥品或準藥品，亦可用作化妝品、醫藥品或準藥品之成分。化妝品、醫藥品或準藥品例如可為溶液、乳液、軟膏、乳霜、糊劑、啫喱、氣溶膠、粉體、油、固形、泡沫等形態。上述發酵物可藉由滴加、散佈、塗抹等而直接應用於皮膚，亦可進行經口投予。本實施形態之發酵物可為皮膚外用劑。作為上述化妝品或準藥品，例如可列舉化妝水、洗面乳、乳液、乳霜、凝膠、精華、美容液、敷劑(pack)、面膜(mask)、化妝油、軟膏等基礎化妝品，粉餅、口紅、胭脂、眼影、眼線、睫毛膏、白粉等彩妝化妝品，洗面乳、按摩用劑、清潔用劑、鬍後乳液、鬍前乳液、刮鬍膏、沐浴乳、身體乳霜、肥皂、洗髮精、潤絲精、護髮素、日曬乳液、防曬乳液、日曬乳霜、防曬乳霜、日曬凝膠、防曬凝膠、整髮劑、養髮劑、生髮劑、止汗劑、沐浴劑、口腔清洗劑、漱口水等。 於將本實施形態之發酵物用作化妝品、醫藥品或準藥品之成分之情形時，發酵物之含量例如可為化妝品、醫藥品或準藥品中之0.01～100%，較佳為0.1～50%，更佳為0.3～3.0%，進而較佳為0.4～1.5%。 於將本實施形態之發酵物用作醫藥品或準藥品之成分之情形時，該醫藥品或準藥品例如亦可包含藥學上容許之成分(例如，賦形劑、結合材、潤滑劑、崩解劑、乳化劑、界面活性劑、基劑、溶解助劑、懸浮劑)作為其他成分。 醫藥品或準藥品可為固體、液體、糊劑等之任一形狀，亦可為錠劑(包含素錠、糖衣錠、發泡錠、膜衣錠、咀嚼錠、口含劑等)、膠囊劑、丸劑、粉末劑(散劑)、細粒劑、顆粒劑、液劑、懸浮液、乳濁液、糖漿、糊劑、注射劑(包含於使用時調配至蒸餾水或者胺基酸輸液或電解質輸液等輸液而製備成液劑之情形)等劑形。該等各種製劑例如可藉由混合作為有效成分之上述發酵物與視需要使用之其他成分，成形為上述劑形而製備。 本實施形態之發酵物可直接用作食品，亦可添加至食品而使用。可藉由攝取上述發酵物而期待上述皮膚改善。作為食品，較佳為強調食品之3次功能(健康狀態調節功能)者。作為強調食品之3次功能之食品，例如可列舉健康食品、功能性食品、營養組合物(nutritional composition)、營養輔助食品、補充品、保健用食品、特定保健用食品、營養功能食品、或功能性標示食品等。 於將上述發酵物添加至食品而使用之情形時，食品亦可包含作為食品所容許之成分。關於作為食品所容許之成分，例如可列舉礦物質類、維生素類、類黃酮類、醌類、多酚類、胺基酸、核酸、必需脂肪酸、清涼劑、結合劑、甜味料、崩解劑、潤滑劑、著色料、香料、穩定劑、防腐劑、緩釋調整劑、界面活性劑、溶解劑、濕潤劑等。 作為食品，可列舉如下者，可於該等之製造步驟中之半成品、或最終製品中混合上述發酵物而獲得用於上述目的之食品：咖啡、果汁及茶飲料等清涼飲料、乳飲料、乳酸菌飲料、酸乳酪飲料、碳酸飲料、以及日本酒、洋酒、果酒及蜂蜜酒等酒等飲料；卡士達奶油等塗醬；果糊等糊；巧克力、甜甜圈、餡餅、奶油泡芙、口香糖、果凍、糖果、曲奇、蛋糕及布丁等西式點心；大福餅、年糕、饅頭、長崎蛋糕、豆沙水果涼粉及羊羹等日式點心；冰淇淋、冰棒及冰雪泥等冷凍點心；咖喱、牛肉蓋飯、雜燴粥、味噌汁、湯、肉醬、麵食、醃菜、果醬等經調理之食品；調味醬、香松、鮮味調味料及湯料等調味料。 本實施形態之由發酵物所構成之食品、化妝品、醫藥品或者準藥品、或包含該發酵物之食品、化妝品、醫藥品或者準藥品可為選自由皮膚障壁功能改善用、皮膚保濕用、皮膚之彈力改善用、皮膚之暗沈改善用、皮膚之皺紋改善用、肌膚粗糙改善用、皮膚之老化防止用、肌膚之肌理改善用及敏感肌膚改善用所組成之群中之至少一種。又，該食品、化妝品、醫藥品或準藥品可為緊連蛋白產生促進用、密連蛋白-1產生促進用、密連蛋白-4產生促進用、緊密連接蛋白-1產生促進用、緊密連接蛋白-2產生促進用、緊密連接蛋白-3產生促進用、III型膠原蛋白α1鏈產生促進用、IV型膠原蛋白產生促進用、水孔蛋白3產生促進用、絲胺酸棕櫚醯轉移酶2產生促進用及/或TIMP2產生促進用，且可為編碼該等蛋白質之基因之表現促進用。又，上述食品、化妝品、醫藥品或準藥品可為介白素-1α產生抑制用、介白素-1β產生抑制用、環加氧酶2產生抑制用及/或基質金屬蛋白酶-9產生抑制用，亦可為編碼該等蛋白質之基因之表現抑制用。 於本實施形態之由發酵物所構成或包含該發酵物之食品、化妝品、醫藥品或準藥品例如亦可附有緊密連接之改善；細胞間隙路徑之透過性改善；美化肌膚；皮膚之保濕；皮膚之彈力或緊致感下降、皮膚之乾燥、肌膚粗糙、皺紋、老化之防止；肌膚之緊致；肌膚之肌理、暗沈之改善；敏感肌膚之改善；光老化之防止等標示。再者，所謂光老化之防止具體係指預防或改善因紫外線等光之影響引起之黃褐斑、皺紋等症狀。 實施例 以下，基於實施例而更具體地對本發明進行說明。然而，本發明並不限定於以下之實施例。 [蜂蜜及德國洋甘菊花粉末之發酵液之製備] 製備包含10%(w/w)之羅馬尼亞產洋槐蜂蜜(山田養蜂場公司)、1%(w/w)之德國洋甘菊花粉末之水溶液(pH值為5)。將該水溶液於110℃下處理5分鐘進行殺菌而獲得發酵前液。 於MRS瓊脂培養基中，於30℃下將昆氏乳桿菌BPS402靜置培養48小時。自所形成之菌落用一白金耳植菌至發酵前液50 mL，於30℃下振盪培養24小時，藉此獲得預培養液。 將1 mL之上述預培養液接種至上述含發酵基質之液體1000 mL，以最佳溫度振盪培養24小時。於將所獲得之發酵液於110℃下處理5分鐘進行殺菌後進行過濾，以成為0.5%之濃度之方式添加苯氧基乙醇而獲得試驗用蜂蜜及德國洋甘菊花粉末之發酵液。 [DNA(Deoxyribonucleic Acid，脫氧核糖核酸)微陣列試驗] 使用人類皮膚三維模型套組(EFT-412，Kurabo公司製造)進行DNA微陣列試驗。根據操作流程，將皮膚模型移至6孔培養盤，使用分析培養基以37℃於5% CO2
下進行一夜之預培養。於預培養後，將200 μL之上述發酵液添加至角質層側，於真皮側添加2.5 ml之分析培養基而繼續進行3天培養。於培養結束後，藉由PBS(Phosphate Buffered Saline，磷酸鹽緩衝液)(-)清洗皮膚組織，使用Derma-Punch(直徑為8 mm)回收組織，之後實施DNA微陣列分析。使用添加有苯氧基乙醇0.5%之純化水作為對照組而同樣地進行分析。又，藉由MTT(Methyl Thiazolyl Tetrazolium，甲基噻唑基四唑鎓鹽)法評價受檢物質對皮膚三維模型之細胞毒性。 對42450個基因進行網羅性分析，結果與對照組相比，藉由發酵液處理而表現上升至1.5倍以上之基因為3012個基因，減少至2分之1以下之基因為2826個基因。自其中如表1般抽選編碼15個蛋白質之基因作為與皮膚功能有關之基因。未確認到由上述發酵液產生之細胞毒性。 [表1]
如表1所示，於蜂蜜及德國洋甘菊花粉末發酵液中，編碼構成緊密連接之蛋白質、膠原蛋白、水孔蛋白3、腦醯胺合成速率限制酶等參與肌膚之水分保持或障壁功能之提高之蛋白質的基因確認到與純化水相比1.5倍以上之有意義之表現上升。另一方面，對於編碼參與炎症反應之介白素1、報告有因紫外線而上升之環加氧酶-2、及分解構成皮膚基底膜之蛋白質之MMP-9之各基因，確認到蜂蜜及德國洋甘菊花粉末發酵液有意義地抑制表現。 [蛋白質產生促進試驗] 作為受驗物質，使用上述蜂蜜與德國洋甘菊花粉末發酵液、及10%蜂蜜水(羅馬尼亞產洋槐蜂蜜10%、純化水90%)。進而，將該等受驗物質之濃度調整為1.0%或0.5%之水溶液用於以下之試驗。 ＜緊連蛋白產生促進試驗＞ 於使用正常人類表皮角化細胞增殖培養基(KGM)培養正常人類新生兒表皮角化細胞(NHEK)後，藉由胰蛋白酶處理而回收細胞。於以成為2×105
cells/ml之濃度之方式藉由KGM稀釋所回收之細胞後，以每孔(well)100 μL接種至96孔培養盤並培養一夜。於培養結束後，在各孔中添加100 μL之藉由KGM溶解之受驗試樣並培養24小時。於培養結束後，去除培養基，將細胞固定至培養盤，藉由使用多株抗人類緊連蛋白抗體之ELISA(Enzyme Linked Immunosorbent Assay，酵素結合免疫吸附分析)法測定於細胞表面表現之緊連蛋白之量。緊連蛋白產生促進率之計算方法如下。將結果示於表2。 緊連蛋白產生促進率(%)＝A/B×100 A：添加受驗試樣時之波長405 nm下之吸光度 B：未添加受驗試樣時(對照組)之波長405 nm下之吸光度 [表2]
於蜂蜜及德國洋甘菊花粉末發酵液中，在0.5%及1.0%之濃度下發現較高之緊連蛋白產生促進作用。 ＜密連蛋白-4產生促進試驗＞ 於使用正常人類表皮角化細胞增殖培養基(KGM)培養正常人類新生兒表皮角化細胞(NHEK)後，藉由胰蛋白酶處理而回收細胞。於以成為2×105
cells/ml之濃度之方式藉由KGM稀釋所回收之細胞後，以每孔100 μL接種至96孔培養盤並培養一夜。於培養結束後，於各孔中添加100 μL之藉由KGM溶解之受驗試樣並培養24小時。於培養結束後，去除培養基，將細胞固定至培養盤，藉由使用單株抗人類密連蛋白-4抗體之ELISA法測定於細胞表面表現之密連蛋白-4之量。密連蛋白-4產生促進率之計算方法如下。將結果示於表3。 密連蛋白-4產生促進率(%)＝A/B×100 A：添加受驗試樣時之波長405 nm下之吸光度 B：未添加受驗試樣時(對照組)之波長405 nm下之吸光度 [表3]
於蜂蜜及德國洋甘菊花粉末發酵液中，在0.5%及1.0%之濃度之濃度下發現密連蛋白-4產生促進作用。 ＜緊密連接蛋白-2產生促進試驗＞ 於使用正常人類表皮角化細胞增殖培養基(KGM)培養正常人類新生兒表皮角化細胞(NHEK)後，藉由胰蛋白酶處理而回收細胞。於以成為2×105
cells/ml之濃度之方式藉由KGM稀釋所回收之細胞後，以每孔100 μL接種至96孔培養盤並培養一夜。於培養結束後，在各孔中添加100 μL之藉由KGM溶解之受驗試樣並培養24小時。於培養結束後，去除培養基，將細胞固定至培養盤，藉由使用多株抗人類ZO-2抗體之ELIZA法測定於細胞表面表現之ZO-2之量。再者，緊密連接蛋白-2亦稱為ZO-2。ZO-2產生促進率之計算方法如下。將結果示於表4。 ZO-2產生促進率(%)＝A/B×100 A：添加受驗試樣時之波長405 nm下之吸光度 B：未添加受驗試樣時(對照組)之波長405 nm下之吸光度 [表4]
於蜂蜜及德國洋甘菊花粉末發酵液中，發現0.5%濃度下之ZO-2產生促進作用。 如上所述，蜂蜜及德國洋甘菊花粉末發酵液於0.5%及1.0%之濃度下發現緊連蛋白及密連蛋白-4之產生促進作用，於0.5%之濃度下發現ZO-2之產生促進作用。另一方面，10%蜂蜜水於1.0%之濃度下增加緊連蛋白之產生，但未對密連蛋白-4及ZO-2之產生量產生影響。
[主張利用生物材料1] [寄存國家]JP日本 [寄存機構]獨立行政法人製品評價技術基盤機構特許生物寄託中心(International Patent Organism Depositary(IPOD), National Institute of Advanced Industrial Science and Technology(AIST)) [寄存日期]2011/10/03 [寄存號碼]FERM BP-11439
Claims (10)
- 一種發酵物，其係藉由昆氏乳桿菌(Lactobacillus kunkeei)發酵蜂蜜與德國洋甘菊(Matricaria chamomilla L.)或其處理物而獲得。
- 如請求項1之發酵物，其中上述德國洋甘菊或其處理物為德國洋甘菊花之乾燥粉末。
- 如請求項1或2之發酵物，其中上述蜂蜜與上述德國洋甘菊或其處理物之質量比為26：1～4：1。
- 如請求項1至3中任一項之發酵物，其係選自由皮膚障壁功能改善用、皮膚保濕用、皮膚之彈力改善用、皮膚之暗沈改善用、皮膚之皺紋改善用、肌膚粗糙改善用、皮膚之老化防止用、肌膚之肌理改善用及敏感肌膚改善用所組成之群中之至少一種。
- 化妝品、醫藥品或準藥品，其包含如請求項1至4中任一項之發酵物。
- 一種發酵物之製造方法，其包括藉由昆氏乳桿菌(Lactobacillus kunkeei)發酵蜂蜜與德國洋甘菊(Matricaria chamomilla L.)或其處理物而獲得發酵物之步驟。
- 如請求項6之製造方法，其中上述德國洋甘菊或其處理物為德國洋甘菊花之乾燥粉末。
- 如請求項6或7之製造方法，其中上述蜂蜜與上述德國洋甘菊或其處理物之質量比為26：1～4：1。
- 如請求項6至8中任一項之製造方法，其中上述發酵物係選自由皮膚障壁功能改善用、皮膚保濕用、皮膚之彈力改善用、皮膚之暗沈改善用、皮膚之皺紋改善用、肌膚粗糙改善用、皮膚之老化防止用、肌膚之肌理改善用及敏感肌膚改善用所組成之群中之至少一種。
- 化妝品、醫藥品或準藥品之製造方法，其包括添加藉由如請求項6至9中任一項之製造方法而獲得之發酵物之步驟。
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| CN110446786A (zh) * | 2016-12-28 | 2019-11-12 | 株式会社山田养蜂场本社 | 蜂蜜发酵物及其制造方法 |
| WO2020090321A1 (ja) | 2018-10-30 | 2020-05-07 | 株式会社村田製作所 | 熱ショックタンパク質遺伝子発現調整剤、医薬品、化粧品、及び熱ショックタンパク質遺伝子発現調整剤の製造方法 |
| CN114450019A (zh) | 2019-09-25 | 2022-05-06 | 株式会社村田制作所 | 胶原蛋白产生促进剂、药物、化妆品和胶原蛋白产生促进剂的制造方法 |
| JP7481748B2 (ja) | 2021-01-29 | 2024-05-13 | 株式会社山田養蜂場本社 | サーカディアン・リズム改善剤 |
| KR102453319B1 (ko) * | 2021-08-09 | 2022-10-12 | 주식회사 한국화장품제조 | 락토바실러스 쿤키 Hankook-001 균주를 이용한 항산화, 항염, 미백, 항주름용 발효추출물 및 그 제조 방법, 그 발효추출물을 포함하는 항산화, 항염, 미백, 항주름용 화장료 조성물 |
| CN113679655B (zh) * | 2021-08-26 | 2023-01-31 | 浙江辰海生命科学有限公司 | 蜂蜜发酵物、含其皮肤外用剂及其制备方法和应用 |
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| JPH07187991A (ja) * | 1993-12-27 | 1995-07-25 | Taisho Pharmaceut Co Ltd | シソエキスおよびカミツレエキス含有浴用剤 |
| US20060018867A1 (en) * | 2004-05-12 | 2006-01-26 | Ichimaru Pharcos Co., Ltd | Cosmetic composition and production thereof |
| JP4982052B2 (ja) * | 2005-06-07 | 2012-07-25 | オリザ油化株式会社 | 美肌用組成物 |
| EP2142635B1 (en) * | 2007-05-03 | 2018-05-23 | Tobias Olofsson | Novel bacteria isolated from the honey producing tract of honey bees |
| JP5860577B2 (ja) | 2008-04-17 | 2016-02-16 | 丸善製薬株式会社 | クローディン産生促進剤及びオクルディン産生促進剤 |
| JP2012121871A (ja) | 2010-12-10 | 2012-06-28 | Shiseido Co Ltd | 皮膚バリア機能改善剤 |
| US9856451B2 (en) * | 2011-12-28 | 2018-01-02 | Yamada Bee Company Inc. | Lactic acid bacterium having IgA production promoting activity, and use thereof |
| JP6125771B2 (ja) * | 2012-07-31 | 2017-05-10 | 株式会社ゼンシン | 化粧料の製造方法、及び、同製造方法により製造した化粧料 |
| CN103564592B (zh) * | 2013-11-21 | 2015-06-24 | 成都冯氏蜂业有限公司 | 一种蜂蜜发酵饮料的生产方法 |
| JP6321402B2 (ja) * | 2014-02-21 | 2018-05-09 | 株式会社ナリス化粧品 | 細胞分化促進剤及び化粧料 |
| JP6486612B2 (ja) * | 2014-05-27 | 2019-03-20 | 株式会社山田養蜂場本社 | 抗アレルギー作用を有する乳酸菌及びその用途 |
| KR101719223B1 (ko) | 2014-10-28 | 2017-03-23 | 주식회사 코리아나화장품 | 김치유산균으로 발효된 인삼, 꿀 및 프로폴리스의 발효액을 함유한 세라마이드 생합성 촉진용 화장료 조성물 |
| CN105076934A (zh) * | 2015-08-31 | 2015-11-25 | 孙培力 | 一种乳酸蜜制作方法 |
| CN105176770A (zh) * | 2015-10-21 | 2015-12-23 | 西安木本贞生物科技有限公司 | 一种含有益菌营养保健的蜂蜜饮料及其制作方法 |
| CN105997815B (zh) * | 2016-07-07 | 2019-03-01 | 广州莱丽奇生物科技有限公司 | 一种多功能美容浓缩液的制备方法 |
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| CN110249055A (zh) | 2019-09-17 |
| WO2018123828A1 (ja) | 2018-07-05 |
| US20190365828A1 (en) | 2019-12-05 |
| EP3564382A4 (en) | 2020-09-02 |
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| CN110249055B (zh) | 2023-01-03 |
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