TW201542215A - 戊糖素(pentosidine)生成抑制劑 - Google Patents
戊糖素(pentosidine)生成抑制劑 Download PDFInfo
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- TW201542215A TW201542215A TW103123148A TW103123148A TW201542215A TW 201542215 A TW201542215 A TW 201542215A TW 103123148 A TW103123148 A TW 103123148A TW 103123148 A TW103123148 A TW 103123148A TW 201542215 A TW201542215 A TW 201542215A
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- Prior art keywords
- lactic acid
- acid bacteria
- pentosidine
- fermentation product
- production inhibitor
- Prior art date
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Abstract
作為具有抑制戊糖素生成之作用的成分,具有更強作用、安全性高之以乳酸菌發酵物為有效成分戊糖素生成抑制劑。
Description
本發明係關於以乳酸菌發酵物為有效成分之戊糖素生成抑制劑。
戊糖素係具有離胺酸殘基與精胺酸殘基以五碳糖交聯之構造的高度糖化終產物(Advanced Glycation End products:AGEs)。
此戊糖素係累積於人類硬膜或皮膚膠原蛋白。又,戊糖素於風濕病、或重症之異位性皮膚炎患者中亦有檢測出高值者,亦顯示了與此等疾病的關聯性。
因此藉由抑制戊糖素之生成,可進行與戊糖素關聯之疾病的預防或治療。
但是,至今為止,作為具有抑制戊糖素生成之作用的成分,雖已知有依達拉奉(edaravone)(專利文獻1)等,但效果或安全性等未能充分達到滿足。
[專利文獻1]日本特開2004-300153號公報
因此,本發明之課題為提供具有更強的作用,且安全性高者,作為具有抑制戊糖素生成之作用的成分。
本發明者等人,為了解決上述課題努力研究的結果,發現了乳酸菌發酵物意外地具有抑制戊糖素生成的作用。又,本發明者等人,發現了於乳酸菌發酵物中,尤其以將含有乳成分或蘆薈之培養基以乳酸菌發酵而得之乳酸菌發酵物為有效成分時,抑制戊糖素之生成的作用較高,而完成了本發明。
亦即,本發明係以乳酸菌發酵物為有效成分之戊糖素生成抑制劑。
又,本發明係以將含有乳成分之培養基以乳酸菌發酵而得之乳酸菌發酵物為有效成分的戊糖素生成抑制劑。
進一步地,本發明係以將含有蘆薈之培養基以乳酸菌發酵而得之乳酸菌發酵物為有效成分的戊糖素生成抑制劑。
又進一步地,本發明係人類中之戊糖素生成
之抑制方法,其特徵為將上述以乳酸菌發酵物為有效成分之戊糖素生成抑制劑對人類投與。
本發明之戊糖素生成抑制劑,即使於低濃度其作用亦強,而且係來自天然物,因此安全性高,可進行與戊糖素關聯之疾病的預防、治療。
本發明之戊糖素生成抑制劑(以下稱為「本發明抑制劑」),係以乳酸菌發酵物為有效成分者。本說明書中,乳酸菌發酵物係指將能夠以乳酸菌發酵之原料以乳酸菌發酵而得者。又,乳酸菌發酵物可含有乳酸菌、亦可不含有乳酸菌,亦包含藉由過濾等,由乳酸菌之培養物中去除乳酸菌後的上清等。
又,本說明書中,戊糖素生成抑制,係指減少所生成之戊糖素的量者,戊糖素生成抑制率,係指根據論文(D.R.Sell and V.M.Monnier,J.Biol.Chem 264,21597-21602,(1989))記載之方法,於等量混合有核糖、離胺酸鹽酸鹽、精胺酸鹽酸鹽之基質溶液中,藉由測定、比較被測物質存在下或非存在下所生成之戊糖素的量所算出的值。更具體而言,係於等量混合有核糖、離胺酸鹽酸鹽、精胺酸鹽酸鹽之基質溶液中混合被測物質,於60℃反應24小時後,測定照射330nm之激發光所產生的
390nm之螢光強度,由實施例所記載之數1的式子算出之值。
作為乳酸菌發酵物之原料,只要係能夠以乳酸菌發酵者則無特殊限定,可列舉例如含有蘆薈、豆乳等植物由來的原料;或牛乳、人乳、山羊乳等之獸乳、乳脂、脫脂粉乳等之動物由來的原料之培養基、MRS(Man,Rogosa and Sharpe)培養基或M-17培養基等乳酸菌用培養基等。此等原料之中,特佳為使用含有牛乳、人乳、山羊乳等之獸乳、乳脂、脫脂粉乳等之乳成分的培養基;或含有蘆薈之培養基。又,作為此等原料,亦可進行過濾或離心分離處理;溶劑之溶解或稀釋處理;混合器等之粉碎處理;澱粉酶、纖維素酶、果膠酶、蛋白酶等之酵素處理;溶劑之萃取處理。
又,發酵所用之乳酸菌亦無特殊限定,可列舉例如乾酪乳酸桿菌(Lactobacillus‧casei)、卷曲乳酸桿菌(Lactobacillus‧crispatus)、胚芽乳酸桿菌(Lactobacillus‧plantarum)、噬澱粉乳酸桿菌(Lactobacillus‧amylovorus)、短乳酸桿菌(Lactobacillus‧brevis)、布氏乳酸桿菌(Lactobacillus‧buchneri)、發酵乳酸桿菌(Lactobacillus‧fermentum)、馬里乳酸桿菌(Lactobacillus‧mali)、類布氏乳酸桿菌(Lactobacillus‧parabuchneri)、副乾酪乳酸桿菌(Lactobacillus‧paracasei)、戊糖乳酸桿菌(Lactobacillus‧pentosus)、雷曼氏乳酸桿菌(Lactobacillus‧
rhamnosus)、犢乳酸桿菌(Lactobacillus‧vitulinus)、玉米乳酸桿菌(Lactobacillus‧zeae)等之乳酸桿菌(Lactobacillus)屬;乳酸乳酸球菌(Lactococcus‧lactis)等之乳酸球菌(Lactococcus)屬;腸膜明念珠菌(Leuconostoc‧mesenteroides)、肉色明串珠菌(Leuconostoc‧carnosum)、檸檬明串珠菌(Leuconostoc‧citreum)、硬明串珠菌(Leuconostoc‧gelidum)、乳明串珠菌(Leuconostoc‧lactis)等之白念珠球菌(Leuconostoc)屬;戊糖片球菌(Pediococcus‧pentosaceus)等之小球菌(Pediococcus)屬;糞腸球菌(Enterococcus‧faecalis)、屎腸球菌(Enterococcus‧faecium)等之腸球菌(Enterococcus)屬;融合魏斯氏菌(Weissella‧confusa)、類腸膜魏斯氏菌(Weissella‧paramesenteroides)、綠色魏斯氏菌(Weissella‧viridescens)等之魏斯氏菌Weissella屬;嗜熱鏈球菌(Streptococcus‧thermophilus)等之鏈球菌(Streptococcus)屬等之乳酸菌。此等乳酸菌可使用1種或2種以上。
以乳酸菌發酵含有上述原料之培養基的條件亦無特殊限定,例如,對前述培養基接種乳酸菌0.01~10質量%、較佳為0.1~5質量%,將之於20~45℃、較佳為25~42℃培養1~96小時、較佳為3~96小時即可。
進一步地,上述發酵時,亦可於培養基中添加例如酵母萃取物、綠藻萃取物、維生素類、蛋白分解物、胺基酸類、礦物質類、鹽類、界面活性劑、脂肪酸、
金屬類等。
關於以上述方式所得之乳酸菌發酵物,可直接作為本發明抑制劑使用,亦可使用進一步經周知之過濾、透析、沈澱、離心分離等之精製、分離處理;或進一步經溶劑等之萃取處理、加熱處理者。又,亦可施以冷凍乾燥或濃縮乾燥。
如此所得之乳酸菌發酵物,戊糖素生成抑制活性高,乳酸菌發酵物之濃度以乾燥固體成分計為0.05質量%時,戊糖素生成抑制率為25%以上、較佳為26~60%。
本發明所用之乳酸菌發酵物的較佳態樣,可列舉將含有乳成分之培養基以乳酸菌發酵者、或將含有蘆薈之培養基以乳酸菌發酵者。再者,此處所稱之蘆薈,係不僅指蘆薈本身,亦包含來自蘆薈之成分。將含有乳成分之培養基以乳酸菌發酵者之中,尤以由將含有乳成分之培養基以乳酸菌發酵者中所得之上清較佳;特佳為由將含有乳成分之培養基以乳酸菌發酵者中所得之上清的低分子區分(fraction)、由將含有乳成分之培養基以乳酸菌發酵者中所得之上清的高分子區分。又,將含有蘆薈之培養基以乳酸菌發酵者,特佳為由將含有蘆薈之培養基以乳酸菌發酵者中所得之上清。
以下,說明本發明所用之乳酸菌發酵物的較佳態樣,亦即將含有乳成分之培養基以乳酸菌發酵者、及將含有蘆薈之培養基以乳酸菌發酵者。
使用於將含有乳成分之培養基以乳酸菌發酵者(以下亦有稱為「乳酸菌發酵物(乳成分)」者)的乳酸菌,較佳例如為嗜熱鏈球菌等之鏈球菌(Streptococcus)屬;乾酪乳酸桿菌、胚芽乳酸桿菌等之乳酸桿菌(Lactobacillus)屬等之乳酸菌。此等乳酸菌之中尤以鏈球菌屬之乳酸菌較佳;更佳為嗜熱鏈球菌。又,其中尤以嗜熱鏈球菌YIT2084株(FERM BP-10879、寄存日:2006年8月18日)、嗜熱鏈球菌YIT2085株(FERM BP-10880、寄存日:2006年8月18日)、嗜熱鏈球菌YIT2021株(FERM BP-7537、寄存日:平成8年(1996年)11月1日)、嗜熱鏈球菌YIT2059株(FERM BP-10878、寄存日:2006年8月18日)、嗜熱鏈球菌YIT2001株(FERM BP-7538、寄存日:平成13年(2001年)1月31日)等較佳;特佳為嗜熱鏈球菌YIT2084株(FERM BP-10879、寄存日:2006年8月18日)。再者,此等乳酸菌可使用1種或2種以上。又,上述寄存編號之乳酸菌株,係寄存於獨立行政法人產業技術總合研究所 專利生物寄存中心(日本國 茨城縣筑波市東1丁目1番地1中央第6(郵遞區號305-8566)),但平成25年4月1日起由獨立行政法人製品評估技術基盤機構 專利生物寄存中心(日本國千葉縣木更津市kazusa鎌足2丁目5番地8 120號室(郵遞區號292-0818))所承接。
欲以乳酸菌發酵含有乳成分之培養基,只要遵照周知之培養條件即可,例如,將以水調整為乳成分
1~20質量%者作為培養基,於其中接種乳酸菌0.01質量%~10質量%、較佳為0.1質量%~5質量%,於20℃~45℃、較佳為37℃~42℃,培養1小時~48小時、較佳為4小時~30小時即可。再者,作為此時之其他培養條件,可列舉靜置、攪拌、振盪、通氣等,只要由此等中適當選擇適於培養的方法來進行即可。
又,於含有乳成分之培養基中,亦可於乳酸菌培養中以補充營養源為目的,另外摻合一般所用之成分。作為如此之成分,可列舉酵母萃取物、綠藻萃取物;維生素A、維生素B類、維生素C、維生素E等之維生素類;或包含各種胜肽類之蛋白分解物;胺基酸類;鈣、鎂等之鹽類;聚山梨醇酯80等之界面活性劑;油酸等之脂肪酸;鈣、鎂、錳等之金屬類。
如此方式所得之乳酸菌發酵物(乳成分),亦可施以周知之過濾、透析、沈澱、離心分離等之精製、分離處理;或進一步施以溶劑等之萃取處理、加熱處理、冷凍乾燥或濃縮乾燥。
本發明抑制劑,於乳酸菌發酵物(乳成分)之中,尤以由將含有乳成分之培養基以乳酸菌發酵者中所得之上清較佳;特佳為由將含有乳成分之培養基以乳酸菌發酵者中所得之上清的低分子區分、或由將含有乳成分之培養基以乳酸菌發酵者中所得之上清的高分子區分。
由將含有乳成分之培養基以乳酸菌發酵者中所得之上清(以下亦有稱為「乳酸菌發酵物(乳成分)上
清」者),可藉由自如上述方式所得之乳酸菌發酵物(乳成分),遵照離心分離、過濾等之一般方法,去除固體物而製造。
又,由將含有乳成分之培養基以乳酸菌發酵者中所得之上清的低分子區分(以下亦有稱為「乳酸菌發酵物(乳成分)上清的低分子區分」者),係指上述乳酸菌發酵物(乳成分)上清之分子量20,000Da以下的區分。可藉由對乳酸菌發酵物(乳成分)上清進行超微過濾、凝膠過濾、透析等之處理而採取分子量20,000Da以下的區分。
如此方式所得之乳酸菌發酵物(乳成分)上清的低分子區分,其濃度以乾燥固體成分計為0.05質量%時,戊糖素生成抑制率為25%以上、較佳為26~40%。
又,由將含有乳成分之培養基以乳酸菌發酵者中所得之上清的高分子區分(以下亦有稱為「乳酸菌發酵物(乳成分)上清的高分子區分」者),係指上述乳酸菌發酵物(乳成分)上清之分子量為3,500Da以上的區分。可藉由對乳酸菌發酵物(乳成分)上清進行超微過濾、凝膠過濾、透析等之處理而採取分子量為3,500Da以上的區分。再者,此時,於去除固體物時,較佳為將乳酸菌發酵物中所含之酪蛋白等之蛋白質,預先以三氯乙酸、乙醇等沈澱。藉由預先如此進行,乳酸菌發酵物之上清的高分子區分中會含有多量的多糖類。再者,欲使用含有更多多糖類之高分子區分時,特佳為在將含有乳成分之培養
基以乳酸菌發酵時,於pH6~8之條件下、於37~40℃培養。培養基pH之調整,可使用氫氧化鈉等。
如此方式所得之乳酸菌發酵物(乳成分)上清的高分子區分,其濃度以乾燥固體成分計為0.05質量%時,戊糖素生成抑制率為25%以上、較佳為35~60%。
另一方面,將含有蘆薈之培養基以乳酸菌發酵者(以下亦有稱為「乳酸菌發酵物(蘆薈)」者),如上所述,例如可藉由將乳酸菌以含有蘆薈之培養基培養而得到。
培養基中所含有之蘆薈,係非洲原產之百合科植物的總稱,具有樹蘆薈(Aloe arborescens)或吉拉索蘆薈(Aloe vera)等300種以上之品種。此蘆薈係已知多量含有黏蛋白或蘆薈聚甘露糖等之多糖類,此外亦含有蘆薈素(aloin)或蘆薈寧(aloenin)、蘆薈酊(aloetin)、蘆薈米嗪(alomicin)、蘆薈烏爾辛(aloeulcin)等之成分。本發明中所使用之蘆薈,只要係蘆薈類則無特殊限定,但特別就獲得的容易性、或成本的觀點等而言,較佳為使用樹蘆薈或吉拉索蘆薈。上述蘆薈之使用部位並無特殊限定,可使用葉、葉肉、芽、根之外,因無纖維成分,亦可使用可使之後之製造步驟變得簡便之前述部位的汁液、滴液(drip)、由其所得之各種成分等。
此外,乳酸菌無法代謝利用上述蘆薈中之纖維成分或蛋白成分等,因此較佳為遵照一般方法,於發酵處理前對蘆薈進行纖維素酶處理、蛋白酶處理等之分解處
理等。藉由進行纖維素酶處理,代謝利用性提高,發酵會進行,於乳酸菌發酵物中不會殘存無法代謝利用之纖維素等,之後的製造步驟會變得簡便。又,藉由進行蛋白酶處理,乳酸菌發酵物中不殘留蛋白成分等,因此不產生混濁,發酵後不需進行澄清化步驟。進一步地,藉由合併使用澱粉酶處理,可使乳酸菌之代謝利用性更加提高。酵素分解處理中,可由纖維素酶、果膠酶等之纖維質分解酵素(纖維素酶類)、或酸性蛋白酶、中性蛋白酶、鹼性蛋白酶、木瓜酶、無花果酶、鳳梨酵素等之蛋白酶、胜肽酶、聚甘露醣酶等之蛋白分解酵素等(蛋白酶類)、α-澱粉酶、葡萄糖澱粉酶等之澱粉分解酵素(澱粉酶類)中適當選擇1種或2種以上來使用即可。又,為了不損及乳酸菌發酵物之嗜好性及為了充分代謝利用蘆薈,較佳為於蘆薈中不添加糖類。
上述發酵所用之乳酸菌,較佳為乳酸桿菌(Lactobacillus)屬之乳酸菌、更佳為胚芽乳酸桿菌。再者,此等乳酸菌可使用1種或2種以上。
欲將含有蘆薈之培養基以乳酸菌發酵,只要遵照周知之培養條件即可,例如,於將Brix調整為3.0以上、pH調整為4.2以下之蘆薈萃取物中,接種乳酸菌0.01質量%~10質量%、較佳為0.1質量%~5質量%,將其於20℃~40℃、較佳為25℃~40℃,培養6小時~96小時、較佳為24小時~96小時即可。再者,作為此時之其他培養條件,可列舉靜置、攪拌、振盪、通氣等,只要由
此等中適當選擇適於培養之方法來進行即可。
再者,於含有蘆薈之培養基中,亦可於乳酸菌培養中以補充營養源為目的,另外摻合糖類以外之一般所用之成分。作為如此之成分,可列舉酵母萃取物、綠藻萃取物;維生素A、維生素B類、維生素C、維生素E等之維生素類;或包含各種胜肽類之蛋白分解物、胺基酸類;鈣、鎂等之鹽類;聚山梨醇酯80等之界面活性劑;油酸等之脂肪酸;鈣、鎂、錳等之金屬類。
如此方式所得之乳酸菌發酵物(蘆薈),亦可施以周知之過濾、透析、沈澱、離心分離等之精製、分離處理;或進一步施以以溶劑等之萃取處理、加熱處理、冷凍乾燥或濃縮乾燥。
本發明抑制劑,於乳酸菌發酵物(蘆薈)當中,尤以由將含有蘆薈之培養基以乳酸菌發酵者中所得之上清為特佳。
由將含有蘆薈之培養基以乳酸菌發酵者中所得之上清(以下亦有稱為「乳酸菌發酵物(蘆薈)上清」者),可藉由遵照離心分離、過濾等之一般方法,自如上述方式所得之乳酸菌發酵物(蘆薈)中去除固體物來製造。
如此方式所得之由將含有蘆薈之培養基以乳酸菌發酵者中所得到之上清,其濃度以乾燥固體成分計為0.05質量%時,戊糖素生成抑制率為25%以上、較佳為28~50%。
以上說明之本發明抑制劑,可摻合於化粧品、醫藥品、醫藥部外品等之皮膚外用劑組成物中,作為例如化粧水、乳液、各種乳脂、面膜、美容液等之基礎化粧料;洗髮精、潤絲精、護髮乳、頭髮營養液(hair tonic)、整髮液、髮油、髮乳等之頭髮用製品;入浴劑等之浴用化粧品;粉底、口紅、睫毛膏、眼影等之化妝用化粧品;防曬乳等之特殊化粧品等。作為本發明之戊糖素生成抑制劑之乳酸菌培養上清的摻合量,並無特殊限定,只要依皮膚外用劑組成物之形態等而適當設定即可,較佳以乾燥固體成分換算為0.003質量%~1質量%、較佳為0.015質量%~0.6質量%、特佳為0.03質量%~0.3質量%。
上述皮膚外用劑組成物,可在不妨礙本發明效果的範圍內摻合化粧品、醫藥品、醫藥部外品等之領域中使用之周知成分。作為此等周知成分,可列舉例如水、醇類、油成分、界面活性劑、防腐劑、香料、色素、保濕劑、增黏劑、抗氧化劑、鉗合劑、pH調整劑、發泡劑、紫外線吸收‧散射劑、粉體、維生素類、胺基酸類、抗菌劑、植物萃取物、海藻萃取物、各種藥劑等。
本發明抑制劑,可藉由直接使用上述乳酸菌發酵物,或與周知之醫藥品載體組合並製劑化而予以配製。
進一步地,本發明抑制劑,係以有吃過之經驗的乳酸菌發酵物作為有效成分,因此亦可摻合於各種飲食品中。例如可於乳酸菌發酵物中添加適當的助劑後,使
用慣用之手段,成形為適於食用之形態,例如顆粒狀、粒狀、錠劑、膠囊、膏劑等而供食用,且可添加於各種食品,例如火腿、香腸等之食肉加工食品;魚板、竹輪等之水產加工食品;麵包、甜點等中使用,或添加於水、果汁、牛乳、清涼飲料等之飲料中使用。
以上說明之本發明抑制劑,可藉由對人類的投與,抑制人類中之戊糖素生成,因此可預防或治療與戊糖素關聯之疾病。本發明抑制劑對人類的投與方法並無特殊限定,可列舉例如於人類中生成戊糖素的部位,以適於其之投與路徑,投與可進行上述預防或治療之量即可。生成戊糖素之部位,可列舉例如皮膚、硬膜等。又,適於前述部位之投與路徑,可列舉例如對上述部位之塗佈、經口投與、注射等。
以下列舉實施例以詳細說明本發明,但本發明不受此等實施例之任何限定。
乳酸菌發酵物(乳成分)上清的低分子區分之戊糖素生成抑制活性:
將嗜熱鏈球菌YIT2084株(FERM BP-10879)之-80℃保存菌株,以1接種環植菌於2mL試驗管中之含有10g/L乳糖的M-17培養基(Difco公司製)中,於40℃靜置
培養一夜。接著,將此培養物,以成為1質量%的方式植菌於2mL之10質量%脫脂粉乳水溶液(Difco公司製)中後,於40℃進行靜置培養一夜,作為前培養。將前培養物以1質量%植菌於正式培養培養基(3質量%脫脂粉乳水溶液)100mL中,於40℃靜置培養24小時。將培養後之菌液,於4℃、8,000xg離心分離15分鐘,去除沈澱物。使用區分分子量20,000Da之離心分離方式超微過濾過濾器(Centricut Mini V-20:倉敷紡績公司製),將所得之沈澱物除去液(上清),於4℃、3,000xg進行1小時超微過濾,得到低分子區分10mL。此低分子區分中,係含有乾燥固體成分2%(20,000μg/mL)。又,此乳酸菌發酵物(乳成分)上清的低分子區分之平均分子量為300Da左右。
再者,平均分子量,係將上述低分子區分溶解於50mM氯化鈉溶液後,藉由以下述條件進行HPLC所測定之值。
裝置:Waters-600E
檢測器:ISI RI-980
管柱:ShodexSUGARKS-804
管柱溫度:80℃
移動相:50mM NaCl
流動速度:1mL/分鐘
注入量:10μL
由戊糖素之生成量算出上述所得之低分子區分的戊糖素生成抑制率(%)。首先,準備各等量混合有50mM核糖、50mM離胺酸鹽酸鹽、50mM精胺酸鹽酸鹽及100mM磷酸鈉緩衝液(pH7.4)者,作為基質溶液。其次,於此基質溶液中,混合以100mM磷酸鈉緩衝液(pH7.4)稀釋上述所得之低分子區分而得的被測試樣溶液,使基質溶液:被測試樣溶液成為5:1,於60℃之恆溫槽中反應24小時。使用水以取代低分子區分,同樣地進行反應,作為控制組。再者,使用不含核糖、離胺酸、精胺酸之緩衝液,作為空白組。加熱結束後,對各檢體照射330nm之激發光,測定所產生之390nm之螢光強度,以螢光量算出作為戊糖素之生成量。由此螢光強度,藉由以下式子計算戊糖素生成抑制率。此等結果示於表1。
[數1]戊糖素生成抑制率(%)=((Ec-Eb)-(Es-Eb))/(Ec-Eb)×100
Es:試樣之螢光強度
Ec:控制組之螢光強度
Eb:空白組之螢光強度
由以上結果,可知就乳酸菌發酵物(乳成分)上清的低分子區分而言,以0.05質量%之低濃度,達成高的戊糖素生成抑制率。又,亦確認到戊糖素生成抑制率係濃度依賴性地提高。
乳酸菌發酵物(乳成分)上清的高分子區分之戊糖素生成抑制活性:
將嗜熱鏈球菌YIT2084株(FERM BP-10879)之-80℃保存菌株,以1接種環植菌於2mL試驗管中之含有10g/L乳糖的M-17培養基(Difco公司製)中,於40℃靜置培養一夜。隨後,將此培養物,以成為1質量%的方式植菌於2mL之10質量%脫脂粉乳水溶液(Difco公司製)中後,於40℃進行靜置培養一夜,作為前培養。將此前培養物,以0.1質量%植菌於準備於缸式發酵槽(jar fermenter)中的正式培養培養基(含有1%之大豆胜肽的10質量%脫脂粉乳水溶液)1L中,一邊將培養基使用8N氫氧化鈉溶液保持於pH7,同時在旋轉數100rpm之條件下培養於40℃、24小時。接著,於冰中冷卻後,以終濃
度成為10%w/v的方式添加三氯乙酸(TCA),於4℃靜置2小時。將之以18,300xg離心30分鐘,於上清液中添加等量之99質量%冷乙醇,進一步於4℃靜置一夜。將其以透析管(Spectra/Por Membrane(MW3,500):Spectrum Medical公司製)透析,將所得之高分子區分遵照一般方法予以冷凍乾燥。再者,此乳酸菌發酵物(乳成分)上清的高分子區分之平均分子量為90萬Da左右。平均分子量之測定方法,係與乳酸菌發酵物(乳成分)上清的低分子區分相同。
對於上述所得之乳酸菌發酵物(乳成分)上清的高分子區分,以與實施例1相同方式算出戊糖素生成抑制率(%)。其結果示於表2。
由以上結果,可知就乳酸菌發酵物(乳成分)上清的高分子區分而言,亦以0.05質量%之低濃度,達成高的戊糖素生成抑制率。又,亦確認到戊糖素生成抑制率係濃度依賴性地提高。
乳酸菌發酵物(蘆薈)上清之戊糖素生成抑制活性:
將吉拉索蘆薈(學名:Aloe barbadensis Miller)之冷凍葉子予以解凍後破碎,添加2倍量之水及檸檬酸0.05質量%,於40℃使纖維素酶(纖維素酶“Onozuka”3S:養樂多藥品工業公司製)作用3小時。之後,一時加熱至90℃後,冷卻至室溫並過濾,進行減壓濃縮至成為Brix3.5,得到吉拉索蘆薈萃取液。將所得之吉拉索蘆薈萃取液(pH3.7)於100℃加熱滅菌100分鐘。於其中接種1%之胚芽乳酸桿菌之標準株(ATCC14917)的前培養菌液後,於30℃靜置培養72小時。培養後,以過濾採取濾液,得到乳酸菌發酵物(蘆薈)上清。於此乳酸菌發酵物(蘆薈)上清中,係含有乾燥固體成分2.9%(29,000μg/mL)。
對於上述所得之乳酸菌發酵物(蘆薈)上清,以與實施例1相同方式算出戊糖素生成抑制率(%)。又,對於未經胚芽乳酸桿菌發酵之吉拉索蘆薈萃取液,亦同樣地算出戊糖素生成抑制率(%)。將該等結果示於表3及4。
由以上結果,可知乳酸菌發酵物(蘆薈)上清,相較於未經乳酸菌發酵之吉拉索蘆薈萃取液,以0.05質量%之低濃度,具有1.4倍以上之高的戊糖素生成抑制率。又,亦確認到戊糖素生成抑制率係濃度依賴性地提高。
化粧水之配製:
配製如以下表5記載組成之化粧水。再者,化粧水係於成分7中添加成分1~6,並充分攪拌,藉以配製。
乳液之配製:
配製如以下表6記載組成之乳液。再者,乳液係於成分11中添加成分7、8及10並加溫,於80℃添加成分1~6進行乳化,之後添加成分9並攪拌,冷卻至室溫藉以配製。
乳脂之配製:
配製如以下表7記載組成之乳脂。再者,乳脂係於成分13中添加成分9、10、12並加溫,於80℃添加成分1~8進行乳化,之後添加成分11並攪拌,冷卻至室溫藉
以配製。
本發明之戊糖素生成抑制劑,可利用於與戊糖素關聯之疾病的預防或治療。
Claims (10)
- 一種戊糖素(pentosidine)生成抑制劑,其係以乳酸菌發酵物為有效成分。
- 如請求項1之戊糖素生成抑制劑,其中乳酸菌發酵物之濃度以乾燥固體成分計為0.05質量%時,戊糖素生成抑制率為25%以上。
- 如請求項1或2之戊糖素生成抑制劑,其係使用將含有乳成分之培養基以乳酸菌發酵者,作為乳酸菌發酵物。
- 如請求項1或2之戊糖素生成抑制劑,其係使用由將含有乳成分之培養基以乳酸菌發酵者中所得之上清,作為乳酸菌發酵物。
- 如請求項1或2之戊糖素生成抑制劑,其係使用由將含有乳成分之培養基以乳酸菌發酵者中所得之上清的低分子區分,作為乳酸菌發酵物。
- 如請求項1或2之戊糖素生成抑制劑,其係使用由將含有乳成分之培養基以乳酸菌發酵者中所得之上清的高分子區分,作為乳酸菌發酵物。
- 如請求項3~6中任一項之戊糖素生成抑制劑,其中乳酸菌係嗜熱鏈球菌(Streptococcus‧thermophilus)。
- 如請求項3~6中任一項之戊糖素生成抑制劑,其中乳酸菌係嗜熱鏈球菌(Streptococcus‧thermophilus)YIT2084株(FERM BP-10879)。
- 如請求項1或2之戊糖素生成抑制劑,其係使用將 含有蘆薈之培養基以乳酸菌發酵者,作為乳酸菌發酵物。
- 如請求項9之戊糖素生成抑制劑,其中乳酸菌係胚芽乳酸桿菌(Lactobacillus‧plantarum)。
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US (1) | US10426804B2 (zh) |
JP (1) | JP6326413B2 (zh) |
KR (1) | KR102355751B1 (zh) |
CN (1) | CN105358165A (zh) |
HK (1) | HK1215168A1 (zh) |
MY (1) | MY186332A (zh) |
SG (1) | SG11201509599QA (zh) |
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WO (1) | WO2015002043A1 (zh) |
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US20200270709A1 (en) * | 2017-04-06 | 2020-08-27 | Korea Food Research Institute | Novel Strain Having Activity Of Reducing Advanced Glycation End Products And Use Thereof |
KR102230571B1 (ko) * | 2018-03-22 | 2021-03-22 | 한국식품연구원 | 비형광성 최종당화산물 억제용 조성물 및 이의 용도 |
CN111514077A (zh) * | 2020-04-02 | 2020-08-11 | 珠海中科先进技术研究院有限公司 | 一种戊糖乳杆菌芦荟发酵原浆及其制备方法、保存方法与应用 |
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JPS6153217A (ja) | 1984-08-23 | 1986-03-17 | Rikagaku Kenkyusho | 腎疾患予防治療剤 |
KR970009620B1 (ko) * | 1994-07-25 | 1997-06-17 | 주식회사 태평양 | 유산균 변이주를 이용한 발효 알로에 베라겔의 제조방법 |
KR0154255B1 (ko) * | 1995-08-02 | 1998-10-01 | 이분옥 | 유효 생리 활성 물질이 함유된 무궁화 음료 제조방법 |
JP2000264811A (ja) * | 1999-03-19 | 2000-09-26 | Kanebo Ltd | メイラード反応抑制剤 |
AU2001281880A1 (en) * | 2000-06-28 | 2002-01-08 | Sericausa B.V. | Use of a formulation made of or containing at least one dissimilated milk serum |
AU2001273215A1 (en) * | 2000-07-07 | 2002-01-21 | Case Western Reserve University | Olfactory neuron cultures and method of making and using the same |
JP2002241293A (ja) * | 2001-02-13 | 2002-08-28 | Ichimaru Pharcos Co Ltd | メイラード反応阻害剤 |
JP3795011B2 (ja) * | 2002-12-16 | 2006-07-12 | 株式会社ヤクルト本社 | 保湿剤及びこれを含有する化粧料 |
JP2004300153A (ja) | 2003-03-20 | 2004-10-28 | Kiyoshi Kurokawa | 蛋白修飾物生成抑制剤 |
JP4897229B2 (ja) * | 2005-03-15 | 2012-03-14 | 学校法人 関西大学 | メイラード反応阻害剤 |
JP5160889B2 (ja) * | 2005-06-24 | 2013-03-13 | カルピス株式会社 | 表皮細胞分化及び角化促進剤 |
BRPI0716118B1 (pt) * | 2006-08-30 | 2015-10-06 | Yakult Honsha Kk | Agente anti-ruga. |
JP4990297B2 (ja) * | 2007-02-01 | 2012-08-01 | 株式会社Tkバイオ研究所 | ブドウ果皮・種子乳酸菌発酵物及びそれを利用した医薬 |
US7524504B1 (en) * | 2008-01-14 | 2009-04-28 | Michael Bishop | Anti-glycation hydrolysate of G. lucidum |
WO2010151836A2 (en) * | 2009-06-25 | 2010-12-29 | Adam Vital Iii | Robust tagging systems and methods |
EP2586448B1 (en) * | 2010-06-28 | 2018-02-07 | Kabushiki Kaisha Yakult Honsha | Skin properties improving agent for oral administration |
US8591882B2 (en) * | 2010-08-27 | 2013-11-26 | Kabushiki Kaisha Yakult Honsha | Cytoprotective agent |
KR101205097B1 (ko) * | 2011-03-07 | 2012-11-27 | 우석대학교 산학협력단 | 아토피 피부염 예방 및 개선효과를 나타내는 알로에 발효 및 가수분해 추출물의 제조방법 및 이를 이용하여 제조된 화장품 |
KR20120139916A (ko) | 2011-06-20 | 2012-12-28 | 대한민국(농촌진흥청장) | Igf-1을 함유하는 혈당강하 조성물 |
JP5984512B2 (ja) * | 2011-09-13 | 2016-09-06 | 株式会社ラフィーネインターナショナル | 大豆発酵組成物の利用、及び飲食品 |
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MY186332A (en) | 2021-07-10 |
WO2015002043A1 (ja) | 2015-01-08 |
US10426804B2 (en) | 2019-10-01 |
CN105358165A (zh) | 2016-02-24 |
US20160166622A1 (en) | 2016-06-16 |
JP6326413B2 (ja) | 2018-05-16 |
KR20160029019A (ko) | 2016-03-14 |
JPWO2015002043A1 (ja) | 2017-02-23 |
SG11201509599QA (en) | 2016-02-26 |
HK1215168A1 (zh) | 2016-08-19 |
KR102355751B1 (ko) | 2022-01-26 |
TWI679982B (zh) | 2019-12-21 |
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