TW200826951A - Fermented milk for skin amelioration and/or treatment and method for producing the same - Google Patents
Fermented milk for skin amelioration and/or treatment and method for producing the same Download PDFInfo
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200826951 (1) 九、發明說明 【發明所屬之技術領域】 本發明係關於皮膚改善效果及/或治療效果高之組成 物,與使用其之飲食物及發酵乳,特別係關於含有製造特 性優良的乳酸菌之組成物以及發酵乳。 【先前技術】 以皮膚改善及治療爲目的之組成物及飮食品中,係包 含乳酸菌及其培養物。 關於這些組成物及飲食品之先前技術有如下所述者。 使用特定乳酸菌之異位性皮膚炎的抑制劑及食品(專 利文件1 )。 使用特定乳酸菌之培養物而具有皮膚改善效果之保濕 劑,及有效預防•治療皮膚粗糙與乾燥,且賦予皮膚「光 澤」、「緊緻」之化妝料組成物(專利文件2 )。 爲保持皮膚免疫系統平衡而使用的特定乳酸菌及其培 養物,及含有乳酸菌及其培養物之食品等(專利文件3 ) 〇 藉由使用膠原胜肽與神經醯胺及乳酸菌的組合,可改 善皮膚狀況以外’亦可期待改善便秘、恢復疲勞、減輕肩 頸部酸痛等效果的美容食品(專利文件4)。 如上所述,發現乳酸菌及其培養物具有皮膚改善效果 及治療效果,而事實上自過去即期待其應用於產業上。一 般認爲目前正進行許多具體的硏究,且亦存在實際上已完 -5- 200826951 (2) 成製品化者。 然而,迄今的硏究中,針對乳酸菌與皮膚的關係,或 於乳酸菌中添加膠原胜肽與神經醯胺等所組成之組成物與 皮膚的關係的檢討並不充分。因此,針對因乳酸菌及發酵 乳而具有皮膚改善效果等,目前仍處於缺乏科學性根據的 狀態。 於具有皮膚改善效果等迄今的乳酸菌中,製造發酵乳 時,會有無法形成凝乳,以及凝乳形成前需要極長時間等 情形出現。於此情形下,需分別管理雖然具有皮膚改善效 果等但發酵乳製造特性不佳之乳酸菌,與爲形成凝乳而使 用之製造特性優良的乳酸菌,以及因發酵時間變長,而使 全體之製造時間延長。此時,具有皮膚改善效果等迄今的 乳酸菌並非生菌,而或爲死菌,或爲耐熱性高,被限定的 菌體等。亦即,具有皮膚改善效果等迄今的乳酸菌,並不 適用於發酵乳工業上大量的生產(商業性生產)等。 例如,欲使用號稱具有皮膚改善效果等之乳酸菌進行 發酵乳製造時,大約需要如下所述之製作步驟。亦即爲 (1 )單獨培養具有皮膚改善效果等之乳酸菌。(2 ) 因應情況藉由使用離心分離法及膜分離法,將該乳酸菌( 培養菌體)濃縮至規定之濃度,或爲提高乳酸菌之保存性 ’對培養液進行加熱殺菌,與進行乾燥。(3 )將另一種 形成凝乳之乳酸菌,與其經濃縮後之培養菌體,或經加熱 殺菌及乾燥後之菌體,添加入發酵前之調合乳,並進行混 合培養。 -6 - 200826951 (3) 這些製作步驟的管理十分複雜,且亦增加製k費用及 製造時間,混入雜菌之可能性亦高。另外’亦難以獲得受 大眾喜愛之口味良好的發酵乳。 專利文件i :特開2000-095697號公報 專利文件2 :特開2 0 0 3 - 0 8 1 8 0 8號公報 專利文件3:特表2004-510740號公報 專利文件4 :特開2004-254632號公報 【發明內容】 本發明有鑑於上述先前技術之課題點’以提供因來自 天然物質而具高安全性,皮膚改善效果及/或治療效果高 之組成物,且適於皮膚改善及/或治療用途之組成物爲目 的。另外,亦以使用該組成物,提供皮膚改善效果及/或 治療效果高之飲食品及發酵乳,特別係含有製造特性優良 的乳酸菌之組成物以及發酵乳爲目的。 本發明之發明者們有鑑於上述課題,專心硏究結果’ 自使用於調製發酵乳之乳酸菌,以選出皮膚改善效果及治 療效果高者爲目的,藉由進行in vitro試驗’發現嗜熱鏈 球菌 OLS3059 ( Streptococcus thermophilus OLS3059 )特 別地具有高的皮膚改善效果及/或治療效果,本發明遂至 完成。 亦即,與本申請案之申請專利範圍第1項相關之發明 係一種嗜熱鏈球菌 OLS3059 ( Streptococcus thermophilus OLS 3 05 9 )之用途,其係使用於皮膚改善及/或治療。 200826951 (4) 與申請專利範圍第2項相關之發明係一種皮膚改善用 及/或治療用之組成物,其係含有嗜熱鏈球菌〇LS3〇59( Stfeptoeoecus thermophilus OLS3 05 9 )及 / 或其培養物所 成。 與申請專利範圍第3項相關之發明係如申請專利範圍 第2項之皮膚改善用及/或治療用之組成物,其係進而含 有保加利亞桿囷 OLL1073R-1 ( Lactobacillus delbrueckii subspecies bulgaricus OLL 1 073 R-1)及/ 或其培養物。 與申請專利範圍第4項相關之發明係一種飮食品,其 特徵係含有如申請專利範圍第2項之組成物。 與申請專利範圍第5項相關之發明係一種皮膚改善用 及/或治療用之發酵乳,其係使用保加利亞桿菌( Lactobacillus delbrueckii subspecies bulgaricus )及嗜熱 鍵球囷 OLS3059 ( Streptococcus thermophilus OLS3059) 而調製。 與申請專利範圍第6項相關之發明係如申請專利範圍 第5項之皮膚改善用及/或治療用之發酵乳,其係含有膠 原胜肽及/或神經醯胺。 與申請專利範圍第7項相關之發明係一種製造如申請 專利範圍第5或6項之皮膚改善用及/或治療用發酵乳之 方法,其中於調製原料乳後,添加做爲菌翫之保加利亞桿 菌(Lactobacillus delbrueckii subspecies bulgaricus)及 嗜熱鏈球菌 OLS3059 ( Streptococcus thermophilus 0LS 3 0 5 9 ),塡充於容器使其發酵,形成凝乳後進行冷藏 200826951 (5) 與申請專利範圍第8項相關之發明係一種製造如申請 專利範圍第5或6項之皮膚改善用及/或治療用發酵乳之 方法,其中於調製原料乳後,添加做爲菌酝之保加利亞桿 菌(Lactobacillus delbrueckii subspecies bulgaricus )及 嗜熱鏈球菌 OLS3059 ( Streptococcus thermophilus OL S 3 05 9 ),使其發酵,形成凝乳後,破碎凝乳再塡充於 容器中進行冷藏。 與申請專利範圍第9項相關之發明係一種製造如申請 專利範圍第5或6項之皮膚改善用及/或治療用發酵乳之 方法,其中於調製原料乳後,添加做爲菌翫之保加利亞桿 菌(Lactobacillus delbrueckii subspecies bulgaricus )及 嗜熱鏈球菌 OLS3059 ( Streptococcus thermophilus OLS 3 059 ),使其發酵,形成凝乳後,破碎凝乳後將其冷 卻,塡充於容器中並進行冷藏。 於本發明中,「皮膚改善效果及/或治療效果」,係 指針對皮膚的光澤、緊緻、暗沉、斑點、下垂、毛孔明顯 、皮膚紋理、皴紋、乾燥情形、黏腻感、透明感、泛紅、 妝容持久性、浮腫、黑眼圈明顯、皮疹、保濕、色素沉澱 中至少一種以上的狀態,具有改善效果。 另外,「皮膚改善及/或治療」,係指針對皮膚的光 澤、緊緻、暗沉、斑點、下垂、毛孔明顯、皮膚紋理、皺 紋、乾燥情形、黏腻感、透明感、泛紅、妝容持久性、浮 腫、黑眼圈明顯、皮疹、保濕、色素沉澱中至少一種以上 -9- 200826951 (6) 的狀態,具有改善效果。 進而,「皮膚改善及/或治療」’係包含異位性皮膚 炎等各種過敏性皮膚炎所引起皮膚症狀之改善及抑制發病 〇 另外,「皮膚改善用及/或治療用」係指使前述之皮 膚改善效果及/或治療效果發揮爲目的而使用’特別係指 適於使前述之皮膚改善效果及/或治療效果之用途。 於本發明中,由於將「保加利亞桿菌(Lactobacillus delbrueckii subspecies bulgaricus)」於調製發酵乳時, 與嗜熱鏈球菌 OLS3 059 ( Streptococcus thermophilus OLS 3 05 9 )組合用作爲菌酝,而包含所有具有形成凝乳能 力之乳酸菌。例如自明治乳業股份有限公司所製之原味優 格、硬質優格、軟質優格中所分離出之乳酸菌。 本發明中之嗜熱鏈球菌 OLS3059 ( Streptococcus thermophilus OLS 3 05 9 )係存放於獨立行政法人產業技術 綜合硏究所專利生物存放中心,存放編號爲 FERM P-15487 (識別:Streptococcus thermophilus OLS3059,存 放曰(收領日):平成8年2月29日)之微生物。 自該原存放處基於布達佩斯條約於2006年11月29 日進行存放的轉移,國際存放收領編號係FERM BP- 1 0740 〇 該嗜熱鏈球菌OLS3059之菌體特徵如下所述。 菌落特徵:圓形•凸起•平滑型 (B L平板培養基(無馬血),培養溫度:3 7 °C,培 -10- 200826951 (7) 養時間:48小時) 菌體型態:球菌 革蘭氏染色:陽性 氧氣需求性:好氧性 對糖類的同化作用 核酸糖:—,甘露醇:一,山梨糖醇:—,乳糖:+ ,海藻糖:一,寡糖:一,麥芽糖:—,蜜二糖:一,松 三糖:―,樹膠醛糖:一。 本發明中之保力卩利亞桿菌 OLL 1 073 R-1 (200826951 (1) EMBODIMENT OF THE INVENTION [Technical Field] The present invention relates to a composition having a high skin-improving effect and/or a therapeutic effect, and a food or drink and fermented milk using the same, particularly in terms of containing a manufacturing property. A composition of lactic acid bacteria and fermented milk. [Prior Art] The composition and the food for the purpose of skin improvement and treatment include lactic acid bacteria and cultures thereof. The prior art regarding these compositions and foods and drinks is as follows. Inhibitors and foods for atopic dermatitis using specific lactic acid bacteria (Patent Document 1). A moisturizing agent which has a skin-improving effect by using a culture of a specific lactic acid bacterium, and a cosmetic composition which is effective for preventing and treating the skin from being rough and dry, and imparting "light" and "tightness" to the skin (Patent Document 2). Specific lactic acid bacteria and cultures thereof for maintaining the balance of the skin's immune system, and foods containing lactic acid bacteria and cultures thereof (Patent Document 3) 改善 Skin can be improved by using a combination of collagen peptides with neuropterin and lactic acid bacteria In addition to the situation, it is also expected to improve the constipation, restore fatigue, and reduce the effects of shoulder and neck soreness (Patent Document 4). As described above, it has been found that lactic acid bacteria and cultures thereof have skin-improving effects and therapeutic effects, and in fact, they have been expected to be applied to the industry from the past. It is generally believed that many specific studies are currently being carried out, and there are actually those who have actually finished -5-200826951 (2). However, in recent studies, the review of the relationship between the composition of the lactic acid bacteria and the skin, or the addition of the collagen peptide and the neuropterin to the lactic acid bacteria has not been sufficiently evaluated. Therefore, there is still a lack of scientific evidence for the skin improvement effect due to lactic acid bacteria and fermented milk. In the lactic acid bacteria having the skin improvement effect and the like, when the fermented milk is produced, it is impossible to form the curd, and it takes a long time before the formation of the curd. In this case, it is necessary to separately manage the lactic acid bacteria having poor manufacturing properties of fermented milk, such as a skin-improving effect, and the lactic acid bacteria having excellent manufacturing properties for forming a curd, and the production time of the whole due to the long fermentation time. extend. In this case, the lactic acid bacteria having a skin-improving effect and the like are not bacteria, but are dead bacteria, or have high heat resistance and limited cells. In other words, the lactic acid bacteria having the skin improving effect and the like have not been applied to a large amount of production (commercial production) in the fermented milk industry. For example, in order to produce a fermented milk using a lactic acid bacterium which is said to have a skin improving effect or the like, a production step as described below is required. That is, (1) lactic acid bacteria having a skin improving effect and the like are separately cultured. (2) The lactic acid bacteria (cultured cells) are concentrated to a predetermined concentration or the storage stability of the lactic acid bacteria is increased by the centrifugation method and the membrane separation method, and the culture solution is heat-sterilized and dried. (3) The lactic acid bacteria which form another curd, the cultured cells which have been concentrated, or the cells which have been heat-sterilized and dried are added to the blended milk before fermentation, and mixed and cultured. -6 - 200826951 (3) The management of these production steps is very complicated, and it also increases the cost and manufacturing time, and the possibility of mixing with bacteria is high. In addition, it is difficult to obtain fermented milk with good taste that is popular with the public. Patent Document i: JP-A-2000-095697, Patent Document 2: JP-A-2002-00 8-8 8-8 Patent Document 3: Special Table 2004-510740 Patent Document 4: JP-A-2004-254632 SUMMARY OF THE INVENTION The present invention has been made in view of the above-mentioned problems of the prior art to provide a composition having high safety, a skin improving effect and/or a therapeutic effect due to a natural substance, and is suitable for skin improvement and/or The composition for therapeutic use is for the purpose. In addition, the use of the composition provides foods and fermented milk which have a skin-improving effect and/or a therapeutic effect, and are particularly intended to contain a composition of lactic acid bacteria having excellent manufacturing properties and fermented milk. In view of the above-mentioned problems, the inventors of the present invention have focused on the results of 'inducing streptococcus mutans by performing in vitro tests for the purpose of selecting a lactic acid bacterium for modulating fermented milk to select a skin improving effect and a therapeutic effect. OLS3059 (Streptococcus thermophilus OLS3059) particularly has a high skin-improving effect and/or a therapeutic effect, and the present invention has been completed. That is, the invention relating to the first aspect of the patent application of the present application is a use of Streptococcus thermophilus OLS 3 05 9 for skin improvement and/or treatment. 200826951 (4) The invention relating to the second aspect of the patent application is a skin improving and/or therapeutic composition comprising Streptococcus thermophilus LS3〇59 (Stfeptoeoecus thermophilus OLS3 05 9 ) and/or Cultured. The invention relating to the third aspect of the patent application is the composition for skin improvement and/or treatment of claim 2, which further comprises the Bulgarian rod OLL1073R-1 (Lactobacillus delbrueckii subspecies bulgaricus OLL 1 073 R -1) and / or its culture. The invention relating to the fourth aspect of the patent application is a sputum food characterized by a composition as in the second aspect of the patent application. The invention relating to the fifth aspect of the patent application is a fermented milk for skin improvement and/or treatment which is prepared by using Lactobacillus delbrueckii subspecies bulgaricus and Streptococcus thermophilus OLS3059. The invention relating to the sixth aspect of the patent application is a fermented milk for skin improvement and/or treatment according to claim 5, which comprises a collagen peptide and/or a neurosteroid. The invention relating to the seventh aspect of the patent application is a method for producing a skin-improving and/or therapeutic fermented milk according to claim 5 or 6 of the patent application, wherein after the preparation of the raw milk, the Bulgarian is added as a bacterium. Lactobacillus delbrueckii subspecies bulgaricus and Streptococcus thermophilus 0LS 3 0 5 9 are fermented in a container to form a curd and then refrigerated 200826951 (5) related to item 8 of the patent application The invention relates to a method for producing a skin-improving and/or therapeutic fermented milk according to claim 5 or 6, wherein after the preparation of the raw material milk, Lactobacillus delbrueckii subspecies bulgaricus and a hobby are added. Streptococcus thermophilus OLS3059 (Streptococcus thermophilus OL S 3 05 9 ), fermented to form a curd, and then crushed the curd and then refilled in a container for refrigeration. The invention relating to the ninth aspect of the patent application is a method for producing a skin-improving and/or therapeutic fermented milk according to claim 5 or 6, wherein after the preparation of the raw material milk, the bacterium is added as a bacterium. Lactobacillus delbrueckii subspecies bulgaricus and Streptococcus thermophilus OLS 3 059 are fermented to form a curd. After the curd is broken, it is cooled, filled in a container and refrigerated. In the present invention, "skin-improving effect and/or therapeutic effect" refers to the gloss, firmness, dullness, speckle, sagging, pores, skin texture, crepe, dryness, stickyness, and transparency of the skin. At least one of a state of sensation, redness, long-lasting makeup, edema, conspicuous dark circles, rash, moisturizing, and pigmentation has an improvement effect. In addition, "skin improvement and / or treatment" refers to the skin's luster, firmness, dullness, spots, sagging, obvious pores, skin texture, wrinkles, dryness, sticky feeling, transparency, redness, makeup At least one of the persistence, edema, dark circles, rash, moisturizing, and pigmentation of the -9-200826951 (6) has an improved effect. Further, "skin improvement and/or treatment" includes improvement of skin symptoms caused by various allergic dermatitis such as atopic dermatitis and suppression of onset of symptoms. In addition, "skin improvement and/or treatment" means The purpose of using the skin improving effect and/or the therapeutic effect is to use, in particular, a use suitable for the skin improving effect and/or therapeutic effect described above. In the present invention, "Lactobacillus delbrueckii subspecies bulgaricus" is used in combination with Streptococcus thermophilus OLS3 059 (Streptococcus thermophilus OLS 3 05 9 ) as a bacillus when preparing fermented milk, and contains all formed condensate Lactic acid bacteria with milk ability. For example, the lactic acid bacteria isolated from the original flavor, hard yoghurt, and soft yoghurt made by Meiji Dairy Co., Ltd. The Streptococcus thermophilus OLS 3 05 9 is stored in the patented biological storage center of the Industrial and Technological Research Institute of the Independent Administrative Corporation. The storage number is FERM P-15487 (identification: Streptococcus thermophilus OLS3059, storage 曰(collection date): Microbes of February 29, 2008). Since the original deposit was transferred based on the Budapest Treaty on November 29, 2006, the international storage collection number is FERM BP- 1 0740 菌 The bacterial characteristics of the Streptococcus thermophilus OLS3059 are as follows. Colony characteristics: round • convex • smooth type (BL plate medium (no horse blood), culture temperature: 3 7 °C, culture-10-200826951 (7) Feeding time: 48 hours) Bacterial type: cocci leather Lan's staining: positive oxygen demand: aerobic assimilation of sugars nucleic acid sugar: -, mannitol: one, sorbitol: -, lactose: +, trehalose: one, oligosaccharide: one, maltose: - , honey disaccharide: one, pine trisaccharide: ―, gum aldose: one. In the present invention, Bacillus licheniformis OLL 1 073 R-1 (
Lactobacillus delbrueckii subspecies bulgaricus 0LL 1 073 R-l )係存放於獨立行政法人產業技術綜合硏究 所專利生物存放中心,收領編號爲FERM P- 1 7227 (識別 : Lactobacillus delbrueckii subspecies bulgaricus OLL1073 R-1,存放日(收領日):平成11年2月19日 )之微生物。 自該原存放處基於布達佩斯條約於2006年11月29 曰進行存放的轉移,國際存放收領編號係FERM ΒΡ-1〇741 〇 該保加利亞桿菌〇 L L 1 0 7 3 R-1之菌體特徵如下所述。 菌落特徵:不透明·粗糙型 (BL洋菜培養基,培養溫度:37°C,培養時間:48 小時) 菌體型態:桿菌 革蘭氏染色:陽性 -11 - 200826951 (8) 氧氣需求性:兼氣性厭氧性 對糖類的同化作用 核酸糖·一’甘露醇:一,山梨糖醇:一,乳糖:+ ,海藻糖:—,寡糖:一, 麥芽糖:一,蜜二糖:一,松三糖:一,樹膠醛糖: - Ο 本發明可提供使用於皮膚改善及/或治療之嗜熱鏈球 困 OLS3 059 ( Streptococcus thermophilus OLS3 05 9 )之用 途。 根據本發明可提供因來自天然物質而具高安全性,且 適合用於皮膚改善效果及/或治療用途之組成物。另外, 使用該組成物,可提供皮膚改善效果及/或治療效果高之 飮食品及發酵乳,特別係含有製造特性優良的乳酸菌之組 成物以及發酵乳。 根據本發明,對增高整腸(通便)改善效果及/或治 療效果的人,可供皮膚改善效果及/或治療效果高之發酵 乳及其製造方法。 本申請案係提案使用於皮膚改善及/或治療之嗜熱鏈 球菌 OLS3059 ( Streptococcus thermophilus OLS3059 )之 用途。亦即,本發明之發明者們專心硏究結果,自使用於 調製發酵乳之乳酸菌,以選出皮膚改善效果及治療效果高 者爲目的,藉由進行in vitro試驗,發現嗜熱鏈球菌 OLS3059 ( Streptococcus thermophilus OLS3059 )特別地 具有高的皮膚改善效果及/或治療效果’本發明遂至完成 -12- 200826951 (9) 其次,本發明之組成物係皮膚改善用及/或治療用之 組成物,且係含有嗜熱鏈球菌 OLS 3 05 9 ( Streptococcus thermophilus OLS3059)及/或其培養物。本發明之皮膚 改善用及/或治療用之組成物,因來自天然物質而具高安 全性,進而皮膚改善效果及/或治療效果高,且適合用於 這些用途。 本發明之發明者們爲自使用於調製發酵乳,且具有凝 乳形成能力,同時製造特性優良之乳酸菌,選出皮膚改善 效果及治療效果高者,進行in vitro試驗後,如前所述, 發現嗜熱鏈球菌 OLS3059 ( Streptococcus thermophilus OLS 3 05 9 )特別地具有高的皮膚改善效果及/或治療效果 〇 上述本發明之含有嗜熱鏈球菌 OLS 3 059 (Lactobacillus delbrueckii subspecies bulgaricus 0LL 1 073 Rl ) is deposited in the patented biological storage center of the Institute of Industrial Technology, Independent Legal Entity, and is numbered FERM P- 1 7227 (identification: Lactobacillus delbrueckii subspecies bulgaricus OLL1073 R-1, storage day ( Collection date): Microbes of February 19, 2011). Since the original deposit was transferred based on the Budapest Treaty on November 29, 2006, the international storage collection number is FERM ΒΡ-1〇741. The bacterial characteristics of the Bulgarian bacillus LL 1 0 7 3 R-1 are as follows: Said. Colony characteristics: opaque and rough (BL vegetable medium, culture temperature: 37 ° C, culture time: 48 hours) Bacterial type: Bacillus Gram stain: positive-11 - 200826951 (8) Oxygen demand: Gaso-anaerobic assimilation of sugars. Nucleic acid sugar-a-mannitol: one, sorbitol: one, lactose: +, trehalose: -, oligosaccharide: one, maltose: one, honey disaccharide: one, Pine sucrose: I. Gum aldose: - Ο The present invention provides the use of the heat-resistant chain ball OLS3 059 (Streptococcus thermophilus OLS3 05 9 ) for skin improvement and/or treatment. According to the present invention, it is possible to provide a composition which is highly safe from natural substances and which is suitable for use in skin improvement and/or therapeutic use. In addition, it is possible to provide a mash food and a fermented milk which have a high skin improvement effect and/or a therapeutic effect, and particularly a composition containing lactic acid bacteria having excellent production characteristics and fermented milk. According to the present invention, a fermented milk having a skin improving effect and/or a therapeutic effect and a method for producing the same can be provided to a person who has an effect of improving the intestinal tract (stool) and/or a therapeutic effect. This application is intended for use in the skin improvement and/or treatment of Streptococcus thermophilus OLS3059. In other words, the inventors of the present invention focused on the results, and for the purpose of selecting a lactic acid bacterium for fermenting milk to select a skin improving effect and a therapeutic effect, by performing an in vitro test, it was found that Streptococcus thermophilus OLS3059 ( Streptococcus thermophilus OLS3059) particularly has a high skin-improving effect and/or a therapeutic effect. The present invention is completed to -12-200826951. (9) Next, the composition of the present invention is a composition for skin improvement and/or treatment, And it contains Streptococcus thermophilus OLS 3 05 9 (Streptococcus thermophilus OLS3059) and/or its culture. The composition for skin improvement and/or treatment of the present invention has high safety due to natural substances, and thus has a skin-improving effect and/or a therapeutic effect, and is suitable for use in these applications. The inventors of the present invention have been able to select a lactic acid bacterium which has a curd-forming ability and has excellent curative properties, and has a skin-improving effect and a therapeutic effect, and after performing an in vitro test, as described above, Streptococcus thermophilus OLS3059 (Streptococcus thermophilus OLS 3 05 9 ) particularly has a high skin-improving effect and/or therapeutic effect, and the above-described S. thermophilus-containing OLS 3 059 (
Streptococcus thermophilus OLS3059)及 / 或其培養物所 成之皮膚改善用及/或治療用之組成物,可進而含有保加 利亞桿菌 OLL1073R-1 ( Lactobacillus delbrueckii subspecies bulgaricus OLL 1 073 R-1)及 / 或其培養物。 根據發明者們之實驗,發現組合使用嗜熱鏈球菌 Ο L S 3 0 5 9 ( S t r e p t 〇 c 〇 c c u s t h e r m 〇 p h i 1 u s Ο L S 3 0 5 9 )及 / 或 其培養物,與於菌體外生產黏多醣類之保加利亞桿菌 OLL1073R-1 ( Lactobacillus delbrueckii subspecies bulgaricus OLL 1 073R-1)時,皮膚改善效果及/或治療效 果高,且適合用於這些用途。 -13- (10) 200826951 嗜熱鏈球菌 OLS 3 0 5 9係使用於例如保 LB 8 1 ((註冊商標)明治乳業股份有限公司製 前述之嗜熱鏈球菌OLS3 05 9及/或其培養 利亞桿菌 OLL1073R-1 ( Lactobacillus subspecies bulgaricus OLL1073R-1 ),除乳酸 精製後之菌體外,均係包含菌體之培養物、菌 、培養物之上清液、這些物質之濃縮液、濃縮 ,進而因應需要之稀釋液、稀釋物等,及處理 養物所得之全部型態者。 前述之嗜熱鏈球菌OLS3 059,無論活菌及 膚改善效果及/或治療效果高,且可使用於皮 或治療。另外,保加利亞桿菌 OLL 1 073R-1亦 及死菌任一種。進而,嗜熱鏈球菌OLS3059與 菌OLL 1 073 R-1,其菌體之培養法、萃取法、 縮法、乾燥法、稀釋法等均無特別限制。使用 之培養基一般係含有脫脂乳、乳清、乾酪素等 糖類酵母萃取物等,培養方法可適宜地使用一 性或厭氧性之方法,培養溫度可使用例如設定 ’於培養中亦可使用氫氧化鈉等鹼性物質,維 pH在酸性至中性之間,例如維持pH在5〜6 法。除類似之中和培養法之外,尙可使用分批 方法’於進行培養後,亦可因應需求對其培養 進行濃縮、乾燥、稀釋等。進而亦可使用離心 分離法’使培養物之上清液與菌體分離,使菌 加利亞優格 )0 =物,與保加 delbrueckii :菌經分離· 體之萃取物 物、乾燥物 培養液、培 .死菌,其皮 膚改善及/ 可使用活菌 保加利亞桿 分離法、濃 於菌體培養 牛乳蛋白, 般性的好氧 於3 5〜4 5 〇C 持培養基之 之中和培養 培養等培養 物及上清液 分離法及膜 體於濃縮狀 -14- (11) (11)200826951 態下進行回收。因而可藉由對菌體進行超音波處理及酵素 處理等,而萃取出菌體內的成分,以及可對培養物及其上 清液,菌體及其萃取物進行乾燥。亦可將這些物質作爲本 發明前述之組成物之有效成分。 本發明之組成物可添加任意之成分,且可遵循常用方 法使用爲皮膚外用劑。例如可適宜地混合用於一般的化妝 料、藥品、醫藥品等各種成分、油劑、增黏劑、防腐劑、 乳化劑、顏料、pH調整劑、抗氧化劑、香料等,而可適 宜地因應目的而使用於乳霜、乳液、化妝料、粉底、化妝 水、洗髮精等。 本發明之飲食品係含有前述之本發明之組成物者。與 本發明有關之飲食品係具皮膚改善效果及/或治療效果高 ’且適合用於這些用途者。 前述之本發明之組成物,因係使用可用於食用之乳酸 菌爲有效成分,安全無虞,且可作爲飮食品、醫藥品而經 口及/或非經口地投予。 此時,可單獨使用嗜熱鏈球菌 OLS 3 05 9 (The composition for skin improvement and/or treatment of Streptococcus thermophilus OLS3059) and/or its culture may further comprise Bulgarian bacillus OLL1073R-1 (Lactobacillus delbrueckii subspecies bulgaricus OLL 1 073 R-1) and/or culture thereof Things. According to the experiments of the inventors, it was found that a combination of Streptococcus thermophilus LS 3 0 5 9 (S trept 〇c 〇ccustherm 〇phi 1 us Ο LS 3 0 5 9 ) and/or its culture, in vitro When producing the mucopolysaccharide of the Bulgarian bacterium OLL1073R-1 (Lactobacillus delbrueckii subspecies bulgaricus OLL 1 073R-1), the skin improving effect and/or the therapeutic effect is high, and it is suitable for these uses. -13- (10) 200826951 Streptococcus thermophilus OLS 3 0 5 9 is used, for example, in LB 8 1 ((registered trademark) Meiji Dairy Co., Ltd. as described above for Streptococcus thermophilus OLS3 05 9 and/or its culture Bacillus subtilis OLL1073R-1 (Lactobacillus subspecies bulgaricus OLL1073R-1), except for the bacteria after purification of lactic acid, are cultures containing bacteria, bacteria, supernatants of cultures, concentrates of these substances, and concentration. The above-mentioned dilutions, dilutions, etc., and all types of treatments obtained by the treatment of the nutrient. The aforementioned Streptococcus thermophilus OLS3 059, regardless of live bacteria and skin improvement effect and / or therapeutic effect, and can be used for skin or In addition, the Bulgarian bacillus OLL 1 073R-1 is also one of the dead bacteria. Further, the Streptococcus thermophilus OLS3059 and the bacteria OLL 1 073 R-1, the culture method, the extraction method, the shrinking method, the drying method, The dilution method and the like are not particularly limited. The medium to be used generally contains a saccharide yeast extract such as skim milk, whey or casein, and the culture method may suitably use a method of unitropy or anaerobicity, and the culture temperature may be, for example, 'Alkaline substances such as sodium hydroxide can also be used in the culture, and the pH is between acidic and neutral, for example, the pH is maintained at 5 to 6. In addition to the similar medium and the culture method, the batch method can be used. ' After the cultivation, the culture can be concentrated, dried, diluted, etc. according to the demand. Further, the supernatant of the culture can be separated from the cells by centrifugation, so that the bacteria can be added to the cells. = object, and the preservation of delbrueckii: separation of bacteria, body extracts, dried cultures, culture, dead bacteria, skin improvement and / can use live bacteria Bulgarian rod separation method, concentrated in bacterial culture of milk protein, The general aerobic is carried out in the medium of 3 5~4 5 〇C holding medium and culture and culture, and the supernatant separation method and the membrane body are recovered in the state of concentrated-14-(11) (11)200826951. . Therefore, the cells can be extracted by ultrasonic treatment and enzyme treatment, and the culture, the supernatant, the cells and the extract thereof can be dried. These materials can also be used as an active ingredient of the aforementioned composition of the present invention. The composition of the present invention may be added with any component and may be used as a skin external preparation in accordance with a usual method. For example, it can be suitably mixed for various components such as general cosmetics, pharmaceuticals, pharmaceuticals, oils, tackifiers, preservatives, emulsifiers, pigments, pH adjusters, antioxidants, perfumes, etc., and can be suitably adapted It is used for creams, lotions, cosmetics, foundations, lotions, shampoos, etc. The food or drink of the present invention contains the above-described composition of the present invention. The food and drink related to the present invention has a skin-improving effect and/or a therapeutic effect and is suitable for use in such applications. The composition of the present invention described above is safe and effective as a lactic acid bacteria which can be used for eating, and can be administered as a food or a pharmaceutical product orally and/or parenterally. At this time, Streptococcus thermophilus OLS 3 05 9 can be used alone (
Streptococcus thermophilus OLS3059)之菌體,或是混合 於一般日常飲食中而加以使用。另外,亦可分別單獨使用 嗜熱鏈球菌OLS 3 059與保加利亞桿菌OLL 1 073 R-1之菌體 ’或將其組合後混合於一般日常飲食中而加以使用。進而 ,可將嗜熱鏈球菌OLS3059,或嗜熱鏈球菌OLS3059與保 加利亞桿菌0LL 1 073 R-1之組合,使用於製造經口投予劑 及/或非經口投予劑,再與通常的各種輔助劑等混合而進 -15- (12) (12)200826951 行製劑化。 將嗜熱鏈球菌 OLS3059 ( Streptococcus thermophilus OLS 3 05 9 ),或嗜熱鏈球菌 OLS 3 05 9與保加利亞桿菌 OLL 1 0 7 3 R-1組合者,與一般日常飮食成分,及製造經口 投予劑及/或非經口投予劑時所使用之一般的各種輔助劑 等混合使用時,嗜熱鏈球菌0L S 3 05 9之添加量(混合量) 係考慮攝取本發明之飮食品之對象其年齡、體重、投予方 法等,而可適宜地加以決定。例如以1 09 ( 1 0的9次方) cfu/天以上爲佳,101G ( 10的10次方)cfu/天以上更佳 〇 本發明之飮食品係含有前述本發明之組成物之飲料、 錠劑之外,亦可爲餅乾、麵包、海綿蛋糕、發酵乳、優格 、乳酪、奶油、乳瑪琳、鮮奶油、果凍等。本發明之飲食 品的使用型態、使用方法,有直接添加本發明之組成物於 前述之飮食品中之型態、方法。另外,亦有將由此所得本 發明之飮食品之優格、乳酪、奶油等,進而添加於飲食品 中之型態、方法。 因此,本發明之飮食品以使皮膚改善效果及/或治療 效果發揮爲目的,可在使用於一般的飲食品之外,使用方令 特定保健用食品、營養機能食品、健康食品等。 進而,因本發明之飮食品具有皮膚改善效果及/ $ & 療效果,亦可使用於顯示目的爲皮膚改善效果及/或&胃 效果而使用之飮食品。表示方法除可表示於包裝上t ’ 亦包含揭示於廣告單以及宣傳等。 -16- (13) (13)200826951 本發明之組成物及飮食品用作爲醫藥品時,可採用錠 劑、膠囊劑、顆粒劑、散劑、糖漿劑等型態而加以使用。 此時,可於作爲主劑之賦型劑、結合劑、崩解劑、潤滑劑 、矯味劑、矯臭劑、溶解輔助劑、懸浮劑、包覆劑等中, 依據常用方法使用輔助劑而加以製劑化。 本發明之發酵乳係皮膚改善用及/或治療用之發酵乳 ,係使用保加利亞桿菌 (Lactobacillus delbrueckii subspecies bulgaricus )及嗜熱鏈球菌 OLS 3 05 9 (The cells of Streptococcus thermophilus OLS3059) are used in combination with a general daily diet. Alternatively, the cells of Streptococcus thermophilus OLS 3 059 and Bulgarian bacillus OLL 1 073 R-1 may be used alone or in combination and mixed in a general daily diet. Further, the combination of Streptococcus thermophilus OLS3059, or Streptococcus thermophilus OLS3059 and Bulgarian bacillus 0LL 1 073 R-1 can be used for the manufacture of an oral administration agent and/or a parenteral administration agent, and then with the usual Various adjuvants and the like are mixed and formulated into -15-(12) (12)200826951. Combining Streptococcus thermophilus OLS3059 (Streptococcus thermophilus OLS 3 05 9 ), or Streptococcus thermophilus OLS 3 05 9 with Bulgarian bacillus OLL 1 0 7 3 R-1, with general daily foraging ingredients, and manufacturing oral injection When the pre-agent and/or the various auxiliary agents used in the case of the parenteral administration agent are used in combination, the amount (mixing amount) of Streptococcus thermophilus 0L S 3 05 9 is taken into consideration of the ingestion of the food of the present invention. The subject's age, body weight, administration method, and the like can be appropriately determined. For example, it is preferably 1 09 (10 9th power) cfu/day or more, 101G (10th power of 10th) cfu/day or more, and the food of the present invention is a beverage containing the composition of the present invention, In addition to the lozenge, it can also be biscuits, bread, sponge cake, fermented milk, yogurt, cheese, cream, milk marlin, fresh cream, jelly, and the like. The use form and method of use of the food or drink of the present invention include a form and a method of directly adding the composition of the present invention to the aforementioned food. Further, there is a form and method in which the yoghurt, cheese, cream, and the like of the mashed food of the present invention are further added to a food or drink. Therefore, the sputum food of the present invention is intended to provide a skin-improving effect and/or a therapeutic effect, and can be used in addition to general foods and drinks, and can be used for specific health foods, nutritious foods, health foods, and the like. Further, the scorpion food of the present invention has a skin-improving effect and/or a therapeutic effect, and can also be used for a food which is used for the purpose of displaying a skin-improving effect and/or a stomach effect. The representation method can be expressed on the package t ‘ and also included in the advertisement and promotion. -16- (13) (13) 200826951 When the composition of the present invention and the food of the present invention are used as a pharmaceutical, they may be used in the form of a tablet, a capsule, a granule, a powder, or a syrup. In this case, it can be used as an excipient, a binder, a disintegrant, a lubricant, a flavoring agent, a flavoring agent, a dissolution aid, a suspending agent, a coating agent, etc. as a main ingredient, and an auxiliary agent can be used according to a usual method. Formulation. The fermented milk for improving and/or treating fermented milk of the present invention uses Lactobacillus delbrueckii subspecies bulgaricus and Streptococcus thermophilus OLS 3 05 9 (
Streptococcus thermophilus OLS3059)調製而成。 關於本發明之皮膚改善用及/或治療用之發酵乳,因 係使用選自用於調製發酵乳,且具有凝乳形成能力,同時 製造特性優良之乳酸菌中之皮膚改善效果及/或治療效果 高之嗜熱鏈球菌 OLS3059 ( Streptococcus thermophilus OLS 3 05 9 ),並加以調整,故皮膚改善效果及/或治療效 果高。亦即前述本發明之發酵乳係皮膚改善效果及/或治 療效果高,且適於這些用途。 將如前所述之嗜熱鏈球菌OLS 3 05 9與保加利亞桿菌組 合,而調整乳酸菌例如藉由組合二者並使用爲菌酝,可於 較短的時間內形成凝乳。 於前述本發明之皮膚改善用及/或治療用之發酵乳中 ,保加利亞桿菌(Lactobacillus delbrueckii subspecies bulgaricus )可使用爲例如明治乳業股份有限公司所製之 原味優格、硬質優格、軟質優格中所分離出之乳酸菌。這 些藉由組合保加利亞桿菌與嗜熱鏈球菌而調製出之本發明 -17- (14) (14)200826951 的乳酸菌,係風味佳且受消費者喜愛,口感滑順,於運輸 製品時不易使凝乳崩解,具有凝乳穩定之特徵。 前述使用保加利亞桿菌(Lactobacillus delbrueckii subspecies bulgaricus )及嗜熱鏈球菌 OLS3059 (Streptococcus thermophilus OLS3059) is prepared. The fermented milk for skin improvement and/or treatment of the present invention has a skin-improving effect and/or a therapeutic effect in a lactic acid bacterium selected from the group consisting of lactic acid bacteria having a curd-forming ability and having a curd-forming ability and having excellent manufacturing properties. Streptococcus thermophilus OLS3059 (Streptococcus thermophilus OLS 3 05 9 ), and adjusted, so the skin improvement effect and / or treatment effect is high. That is, the fermented milk of the present invention has a high skin-improving effect and/or a therapeutic effect, and is suitable for these uses. The Streptococcus thermophilus OLS 3 05 9 as described above is combined with the Bulgarian bacterium, and the lactic acid bacteria can be adjusted, for example, by combining the two and using the bacterium to form a curd in a shorter period of time. In the above-mentioned fermented milk for skin improvement and/or treatment of the present invention, Lactobacillus delbrueckii subspecies bulgaricus can be used, for example, as the original yoghurt, hard yoghurt, soft yoghurt made by Meiji Dairy Co., Ltd. The lactic acid bacteria isolated. These lactic acid bacteria of the present invention -17-(14)(14)200826951 prepared by combining Bulgarian bacillus and Streptococcus thermophilus have good flavor and are favored by consumers, and have a smooth taste, which is difficult to coagulate when transporting products. The milk disintegrates and has the characteristics of stable milk. The aforementioned use of Lactobacillus delbrueckii subspecies bulgaricus and Streptococcus thermophilus OLS3059 (
Streptococcus thermophilus OLS3059)而調製之本發明之 皮膚改善用及/或治療用之發酵乳,可含有膠原胜肽及/ 或神經醯胺。 如於後述之實施例中所載,將含有膠原胜肽及/或神 經醯胺之本發明之前述發酵乳,與未含之本發明之前述發 酵乳相比較後,發現於含有膠原胜肽及/或神經醯胺之發 酵乳,皮膚改善效果及/或治療效果有較高的傾向。即與 如前所述之僅使用保加利亞桿菌與嗜熱鏈球菌OL S 3 05 9所 調製之本發明之發酵乳相比較,藉由混合膠原胜肽及/或 神經醯胺而使用,可提高皮膚改善效果及/或治療效果。 認爲其係由於皮膚改善效果及/或治療效果高之乳酸菌( 嗜熱鏈球菌 OLS3059 ( Streptococcus thermophilus OLS3 05 9 )),與膠原胜肽及/或神經醯胺混合所產生的 加乘效果。 且自使此效果發揮之觀點,發酵乳中所含之膠原胜肽 以500mg%以上爲佳,700mg%以上更佳,900mg%以上 最佳。發酵乳中所含之神經醯胺,自使前述效果發揮之觀 點,以l〇〇"g%以上爲佳’ 200#g%以上更佳,250//g% 以上最佳。 自皮膚改善效果及/或治療效果的加乘效果之觀點, -18- (15) (15)200826951 於本發明之發酵乳中,除膠原胜肽及/或神經醯胺之外, 亦可添加鞘磷脂、異黃酮素、軟骨素、玻尿酸等使用於飮 食品、醫藥品之具有新陳代謝改善效果的有效成分。 針對以上本發明之皮膚改善用及/或治療用之發酵乳 ,爲尋找影響皮膚改善效果及/或治療效果的因素,以人 類試驗進一步詳細探討後,發現上述本發明之發酵乳,對 於人類具有發酵乳被公認的高整腸效果此一代表性作用外 ,亦具有極高之皮膚改善效果及/或治療效果。 具體而言,使用含有膠原胜肽及/或神經醯胺之本發 明之前述皮膚改善用及/或治療用之發酵乳,與未含之本 發明之前述皮膚改善用及/或治療用之發酵乳,藉由比較 二者之整腸改善效果程度及皮膚改善效果程度,而獲得如 前所述之發現。 即本發明之前述皮膚改善用及/或治療用之發酵乳, 係具有相對於提高整腸(通便)改善效果及/或治療效果 的人,有極高之皮膚改善效果及/或治療效果之特徵。 【實施方式】 實施例 以下爲製造上述本發明之皮膚改善用及/或治療用之 發酵乳之發酵乳製造方法。 於調製原料乳後,添加做爲菌翫之保加利亞桿菌( Lactobacillus delbrueckii subspecies bulgaricus )及嗜熱 鏈球菌 OLS3059 ( Streptococcus thermophilus OLS3059 ) -19- (16) (16)200826951 ,塡充於容器使其發酵,形成凝乳後進行冷藏。 或於調製原料乳後,添加做爲菌酝之保加利亞桿菌( Lactobacillus delbrueckii subspecies bulgaricus )及嗜熱 鏈球菌 OLS3 05 9 ( Streptococcus thermophilus OLS3059 ) ,使其發酵,於形成凝乳後將凝乳破碎,再塡充於容器後 進行冷藏。 又或者,於調製原料乳後,添加做爲菌翫之保加利亞 桿菌(Lactobacillus delbrueckii subspecies bulgaricus ) 及嗜熱鏈球菌 OLS3059 ( Streptococcus thermophilus OLS 3 059 ),使其發酵,形成凝乳後,破碎凝乳後將其冷 卻,塡充於容器中並進行冷藏。 此處之添加做爲菌酝之保加利亞桿菌(Lactobacillus delbrueckii subspecies bulgaricus )及嗜熱鏈球菌 OLS 3 05 9 ( Streptococcus thermophilus OLS3059 )爲止所 進行之調製原料乳,係與過去一般調製發酵乳之步驟相同 ,係調製原料乳,並進行均質化及殺菌•冷卻等處理而調 製原料乳。 製造前述本發明之發酵乳之製造方法中,係使用選自 用於調製發酵乳,且具有凝乳形成能力,同時製造特性優 良之乳酸菌中之皮膚改善效果及/或治療效果高之嗜熱鏈 球菌 OLS3059 ( Streptococcus thermophilus OLS3059), 與保加利亞桿菌此二菌之組合,並將其做爲菌酝。 因此,解決了具皮膚改善效果之迄今之乳酸菌的問題 點(課題點)。過去之乳酸菌雖具有皮膚改善等效果,但 -20 - (17) (17)200826951 由於需分別管理製造特性不佳之乳酸菌,與爲形成凝乳之 製造特性優良之乳酸菌,及發酵時間變長,而引起使全體 製造時間變長之問題。亦即,具有皮膚改善等效果之迄今 的乳酸菌,並不適用於發酵乳之工業上大量的生產(商業 性生產)等。 然而,本發明之發酵乳之製造方法,因解決前述問題 點’而可適用於商業性生產。商業性生產重要係爲非複雜 步驟數。 於前述本發明之發酵乳之製造方法,調製原料乳步驟 中之均質化’可於殺菌前或殺菌後,或於其進行中而施行 〇 且如前所述’使其進行發酵後之冷卻,可於使凝乳形 成後,將凝乳破碎後,或塡充於容器後再進行。 進而,將凝乳破碎後,可添加•混合果肉、蔬菜、各 種醬料及/或各種添加劑。 本發明之發酵乳之製造方法,其發酵時間係可爲用於 製造一般發酵乳的時間,以8小時之內爲佳,6或5小時 以內更佳,4小時以內最佳。 本發明之皮膚改善用及/或治療用之發酵乳,因其風 味極佳,可時常飮用,且可輕鬆地持續攝取。因此,可更 提高其皮膚改善效果及治療效果。 以下舉出與本發明相關之最佳實施例並加以說明,但 本發明並未限定於此。 * 21 - (18) 200826951 〔實施例1〕 (根據in vivo試驗進行異位性皮膚炎抑制效果之驗證) 將雌性4周大之NC/Nga小鼠(japan SLC (股份有限 . )生產),經馴化飼養後,分做下述4群。 投予蒸餾水群(η = 1 0 ) 投予嗜熱鍵球菌OLS3059 ( Streptococcus thermophilus OLS 3 05 9 )(以下亦稱爲 OLS 3 05 9。其係分 離自保加利亞優格LB81(註冊商標)明治乳業股份有限 公司製))群(n=l〇)。 非引發皮膚炎群(n=7) 投予 Lactobacillus rhamnosus ATCC 53103 (以下亦 稱爲ATCC 5 3 1 0 3。)群(係用作與投予OLS 3 05 9群之對 照)(η = 1 〇 )。 使這些小鼠可自由攝取飼料(商品名:CRF-1 ’ Oriental酵母工業(股份有限)製)及蒸餾水。 , 已知ATCC 53103係具有治療異位性皮膚炎效果之乳 酸菌。 除非引發皮膚炎群之外,3試驗群於試驗期間中’以 管餵方式經口進行投予。接著,根據下述方法,自引發皮 膚炎群試驗開始7天後(day 〇 )連續進行。亦即’投予 蒸餾水群於試驗期間(第1天(day -7)〜21天後(day 14 ))中,每日進行經口投予蒸餾水至胃內(lmg/天/ 小鼠),且投與各種乳酸菌群亦進行相同試驗,每日進行 經口投予乳酸菌之水懸浮液至胃內(1 mg/天/小鼠)° -22- ,60 (19) 200826951 各種乳酸菌之水懸浮液係將經加熱處理(溫度7 5 °c 分鐘)後之菌體的凍結乾燥物懸浮於蒸餾水中而製成 種乳酸菌之懸浮液的菌體濃度分別係ATCC 5 3 1 03爲 xl08(10 的 8 次方)cfu/g,OLS3059 爲 8·56χ107( 7 次方)cfu/ g。 試驗開始7天後(day 0 )引起皮膚炎,將症狀 數據化。且於開始引起皮膚炎15天後(day 15 )採 f 清,以 ELISA法測定 IgE濃度。IgE濃度係The fermented milk for skin improvement and/or treatment of the present invention prepared by Streptococcus thermophilus OLS3059) may contain collagen peptide and/or ceramide. As described in the examples to be described later, the fermented milk of the present invention containing the collagen peptide and/or the neuroguanamine is compared with the fermented milk of the present invention not included, and is found to contain the collagen peptide and / or fermented milk of ceramide, the skin improvement effect and / or treatment effect has a higher tendency. That is, compared with the fermented milk of the present invention prepared by using only the Bulgarian bacterium and the Streptococcus thermophilus OL S 3 05 9 as described above, the skin can be improved by mixing the collagen peptide and/or the neuropterin. Improve results and / or treatment effects. It is considered to be a multiplication effect produced by mixing lactic acid bacteria (Streptococcus thermophilus OLS3 05 9 ) with collagen peptide and/or neuropterin due to a skin-improving effect and/or a therapeutic effect. Further, from the viewpoint of exerting this effect, the collagen peptide contained in the fermented milk is preferably 500 mg% or more, more preferably 700 mg% or more, and most preferably 900 mg% or more. The neuroterpene contained in the fermented milk is preferably more than 200% by weight or more, more preferably 250//g% or more, from the viewpoint of exerting the above effects. From the viewpoint of the effect of improving the skin and/or the effect of the therapeutic effect, -18-(15) (15)200826951 may be added to the fermented milk of the present invention in addition to the collagen peptide and/or the neuropterin. Sphingomyelin, isoflavone, chondroitin, hyaluronic acid, and the like are effective ingredients for improving metabolism and improving the metabolism of foods and pharmaceuticals. In view of the above-mentioned fermented milk for improving and/or treating the skin of the present invention, in order to find a factor affecting the skin improving effect and/or the therapeutic effect, the human fermented milk of the present invention is found to have a human Fermented milk is recognized as a highly effective intestinal effect and has a very high skin-improving effect and/or therapeutic effect. Specifically, the above-mentioned skin improving and/or therapeutic fermented milk of the present invention containing a collagen peptide and/or a neuroguanamine, and the aforementioned skin improving and/or therapeutic fermentation of the present invention not included are used. Milk, by comparing the degree of improvement in the effect of the whole intestine and the degree of skin improvement, the findings as described above were obtained. That is, the fermented milk for skin improvement and/or treatment of the present invention has an extremely high skin-improving effect and/or therapeutic effect on a person who improves the effect and/or the therapeutic effect of the whole intestine (stool). Characteristics. [Embodiment] The following is a method for producing fermented milk for producing the above-described skin improving and/or therapeutic fermented milk of the present invention. After the raw milk is prepared, Lactobacillus delbrueckii subspecies bulgaricus and Streptococcus thermophilus OLS3059 -19-(16) (16) 200826951 are added as a bacterium, and the mixture is fermented in a container. After the formation of curd, it is refrigerated. Or after the preparation of the raw milk, Lactobacillus delbrueckii subspecies bulgaricus and Streptococcus thermophilus OLS3059 are added to ferment, and the curd is broken after the curd is formed, and then the curd is broken. Refrigerate after filling the container. Alternatively, after modulating the raw milk, Lactobacillus delbrueckii subspecies bulgaricus and Streptococcus thermophilus OLS 3 059 are added as fermented to form a curd, and after the curd is broken, It is cooled, filled in a container and refrigerated. Here, the raw milk prepared by the addition of Lactobacillus delbrueckii subspecies bulgaricus and Streptococcus thermophilus OLS3059 is the same as the conventional procedure for preparing fermented milk in the past. The raw material milk is prepared by modulating the raw material milk, performing homogenization, sterilization, cooling, and the like. In the method for producing the fermented milk of the present invention, a Streptococcus thermophilus which is selected from the group consisting of a lactic acid bacterium which has a curd-forming ability and has excellent curative properties and/or a therapeutic effect is used. OLS3059 (Streptococcus thermophilus OLS3059), combined with the bacteria of the Bulgarian bacillus, and used as a fungus. Therefore, the problem (problem point) of the lactic acid bacteria having the skin improving effect has been solved. In the past, lactic acid bacteria have effects such as skin improvement, but -20 - (17) (17) 200826951, because it is necessary to separately manage lactic acid bacteria with poor manufacturing characteristics, and lactic acid bacteria having excellent manufacturing properties for forming curd, and the fermentation time becomes longer. This causes problems that make the entire manufacturing time longer. In other words, lactic acid bacteria having hitherto been effective in skin improvement and the like are not suitable for mass production (commercial production) of fermented milk industry. However, the method for producing fermented milk of the present invention can be applied to commercial production because it solves the aforementioned problem. The importance of commercial production is the number of non-complex steps. In the method for producing fermented milk of the present invention, the homogenization in the step of preparing the raw milk can be carried out before or after sterilization, or during the process of carrying out the treatment, and as described above, the fermentation is carried out after fermentation. After the curd is formed, the curd may be broken or may be filled in the container. Further, after the curd is broken, the pulp, vegetables, various sauces, and/or various additives may be added and mixed. In the method for producing fermented milk of the present invention, the fermentation time may be a time for producing a general fermented milk, preferably within 8 hours, more preferably within 6 or 5 hours, and most preferably within 4 hours. The fermented milk for skin improvement and/or treatment of the present invention is excellent in wind odor, can be used from time to time, and can be easily ingested continuously. Therefore, the skin improvement effect and therapeutic effect can be further improved. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Hereinafter, preferred embodiments of the present invention will be described, but the present invention is not limited thereto. * 21 - (18) 200826951 [Example 1] (Verification of inhibitory effect of atopic dermatitis according to in vivo test) Female 4-week-old NC/Nga mice (produced by japan SLC (share limited)) were domesticated After the rearing, the following four groups were divided. The distilled water group (η = 1 0) was administered to Streptococcus thermophilus OLS 3059 (hereinafter also referred to as OLS 3 05 9 ). It was isolated from the Bulgarian Yogurt LB81 (registered trademark) Meiji Dairy Co., Ltd. Ltd.)) group (n=l〇). Non-priming dermatitis group (n=7) was administered Lactobacillus rhamnosus ATCC 53103 (hereinafter also referred to as ATCC 5 3 1 0 3) group (for comparison with administration of OLS 3 05 9 group) (η = 1 〇 ). These mice were allowed to freely take in feed (trade name: CRF-1 'Oriental Yeast Industry Co., Ltd.) and distilled water. ATCC 53103 is known to have lactic acid bacteria for treating atopic dermatitis. The 3 test groups were orally administered by tube feeding during the test period unless a dermatitis group was initiated. Subsequently, it was continued 7 days after the initiation of the skin dermatitis test (day 〇) according to the method described below. That is, 'distilled water group was administered during the test period (day 1 (day -7) to 21 days (day 14)), and oral distilled water was administered daily to the stomach (lmg/day/mouse). The same experiment was carried out on various lactic acid bacteria groups, and an aqueous suspension of lactic acid bacteria was orally administered to the stomach (1 mg/day/mouse). -22-, 60 (19) 200826951 Water suspension of various lactic acid bacteria The liquid system is obtained by suspending the freeze-dried suspension of the cells after being heat-treated (temperature of 75 ° C minutes) in distilled water to prepare a suspension of the lactic acid bacteria to be ATCC 5 3 1 03 as xl08 (10 8th power) cfu/g, OLS3059 is 8.56χ107 (7th power) cfu/g. After 7 days from the start of the test (day 0), dermatitis was caused and the symptoms were digitized. And 15 days after the onset of dermatitis (day 15), the concentration of IgE was determined by ELISA. IgE concentration system
Pharmingen公司製之抗體,且變更該公司所推薦之試 驟之一部分而進行測定。 以下述方法進行調製塵觸破碎液。J Dermatophagoides pterony s sinus 之成蟲(Mite-Dp, 公司製)以無水甲醇進行脫脂後,添加蒸餾水進行超 破碎處理。接著,已離心處理分離水溶性畫分,並將 結乾燥後,以蒸餾水調製蛋白質濃度爲4.5 mg/ ml ; BSA 換算,並以 Bio RAD 公司之 DC protein assay 測 ο \ 引起皮膚炎試驗係變更海野氏等之方法(海野哲 - 其他,過敏 5 0 : 1 1 5 2 - 1 1 6 2,2 0 0 1 )而進行。以剌毛 去引起異位性皮膚炎部位(頭部、外耳部及頸部)之 再以毛刷將SDS溶液(4% )塗佈於全部的引起異位 膚炎部位2 0次。塗佈量2 0次共計約6 0 // 1。待S D S 草乙魚後’於引起皮膚炎試驗群塗佈塵_破碎液,非引 膚炎試驗群塗佈蒸餾水,同樣爲分別塗佈20次。藉 。各 7.13 10的 演變 取血 ί吏用 驗步 !Ρ將 LSL 音波 其凍 (以 定) 史及 刀剃 毛, 性皮 溶液 起皮 由每 -23- (20) 200826951 B重複進行前述之步驟而引起皮膚炎,於試驗開始後7天 (day 〇 ),每日重複進行至試驗終止。 以下述方法測定IgE。即於96孔盤上添加2 // g/ ml -之1次抗體用之抗IgE抗體(Pharmingen公司製),靜置 於 37。。下 1 小時。以 Tween 20/PBS(PBS-Tween,0.05 % )洗淨後,添加BS A/ PBS ( 1% ),靜置於室溫下30 分鐘。再以PBS-Tween洗淨後,添加血清試樣及製作標準 { 曲線之IgE ( Pharmingen公司製),靜置於室溫下30分 鐘。以PBS-Tween洗淨後,添力□ 0.5// g/ml之2次抗體 用之biotin -抗IgE抗體(Pharmingen公司製),靜置於室 溫下 1小時。以 PBS-Tween洗淨後,添加 Streptavidin-HRP ( Pharmingen公司製),靜置於室溫下 30分鐘。 PBS-Tween 洗淨後,添力D TMB + Substrate Chromogen ( DAKO公司製),而開始呈色反應。於呈色反應後加入硫 酸(1N ),以微量盤判讀機測定吸光度(450nm ),而計 I 算出血清中之IgE濃度。 將皮膚炎症狀數據之演變示於圖1。數據化係針對〔1 • 〕形成乾皮•痂皮,〔2〕發紅·出血,〔3〕組織脫落· . 擦傷,〔4〕浮腫,〔5〕搔癢行爲等5個項目,以「0 · 無症狀」,「1·輕度」,「2.中度」,「3.重度」等4 個階段,計算並評價各小鼠所得之各數據。針對引起皮膚 炎開始第13天(day 13 )之症狀數據’投予〇LS3 05 9群 與投予蒸餾水群相比爲有意義的低。反之,投予 ATCC 53 1 03群與投予蒸餾水群相比並無大的差異。 -24- (21) (21)200826951 將總IgE濃度示於圖2。針對引起皮膚炎開始第15天 (day 1 5 )血清中之總IgE,投予OLS3059群與投予蒸餾 水群相比IgE濃度爲有意義的低。反之,投予 ATCC 5 3 1 03群與投予蒸餾水群相比IgE濃度稍低,但不具統計 上之差異。已知ATCC 5 3 1 03爲具有治療異位性皮膚效果 之乳酸菌,但OLS 3 05 9對較異位性皮膚之症狀抑制效果較 ATCC 5 3 1 0 3 爲高。 〔實施例2〕 (根據in vivo試驗進行異位性皮膚炎抑制效果之驗證) 將雌性4周大之NC/Nga小鼠(Japan SLC (股份有限 )生產),經馴化飼養後,分做下述3群。 投予蒸飽水群(n=4) 投予保加利亞桿菌OLL1073R-1 (Lactobacillus delbrueckii subspecies bulgaricus OLL1073R-1,以下亦稱 爲 OLL 1 073 R-1 )群(n= 6 )。 投予嗜熱鏈球菌OLS 3 05 9 ( Streptococcus thermophilus OLS 3 05 9,係分離自保加利亞優格LB81 (明 治乳業(股份有限)公司製)群(η = 5 )。 使這些小鼠可自由攝取飼料(商品名:CRF_1, Oriental酵母工業(股份有限)製)及蒸館水。除引起皮 膚炎期間爲3 0天之外,其餘試驗條件均與施實施例1相 同。 將皮膚炎症狀數據之演變示於圖3。針對引起皮膚炎 -25- (22) (22)200826951 開始第13天(day 13 )之症狀數據,投予OLL 1 073 R-1群 與投予OLS3 05 9群和投予蒸餾水群相比爲有意義的低。 將總IgE濃度示於圖4。針對血清中之總IgE,投予 OLL 1 073 R-1群與投予OLS 3 0 5 9群和投予蒸餾水群相比 IgE濃度爲有意義的低。可知本發明之OLS3 059對異位性 皮膚炎之症狀具抑制效果。另外,亦發現OLL 1 073R-1對 異位性皮膚炎之症狀具抑制效果,但IgE濃度係本發明之 OLS3059 較低。 〔實施例3〕 (根據in vivo試驗進行異位性皮膚炎抑制效果之驗證) 針對異位性皮膚炎之症狀,將發揮毫不遜色於本發明 之 OLS3 05 9之抑制效果且已於實施例 2中確認之 OLL 1 073 R-1,與本發明之0 L S 3 0 5 9組合使用時,對抑制 異位性皮膚炎之症狀進行檢討。 採用二種均分離自保加利亞優格(明治乳業(股份有 限 ) 製 ) 之 Lactobacillus delbrueckii subspecies bulgaricus MEP1705601 ,與 Streptococcus thermophilus MEP 1 705 60 1 之組合,做爲本發明之 OLS 3 05 9 與 OLL 1 07 3R-1組合之比較對象。 將雌性4周大之NC/Nga小鼠(Japan SLC (股份有限 )生產),經馴化飼養後,分做下述4群。 投予以 Lactobacillus delbrueckii subspecies bulgaricus OLL1073R-1 ,與 Streptococcus thermophilus -26- (23) (23)200826951 OLS 3 05 9所調製之發酵物之群(以下亦稱做〇LL1〇73R] + OLS3 05 9 投予群)(n= 8 )。 投予以 Lactobacillus delbrueckii subspecies bulgaricus MEP 1 705 60 1 ’ 與 Streptococcus thermophilus MEP 1 705 60 1所調製之發酵物之群(以丁亦稱做lb MEP 1705601+STMEP 1705601 投予群)(n=9)。 未發酵物之投予群(n=9),及非引起皮膚炎且未發 酵物之投予群(以下亦稱做非引起群)(n二4 )。 發酵物及未發酵物之攝取量均爲lOOmg / mouse / day ,至試驗終止日爲止每日連續經口投予。 發酵物之菌數分別係OLL 1 073 R-1爲lxl〇9cfu/g, OLS3 05 9 爲 3xl 09 9 cfu/g,LB MEP 1 70560 1 爲 8·6χ 108cfu/g,ST ΜΕΡ 1705601 爲 2.1xl〇9cfu/g。 將發酵物及未發酵物凍結乾燥後,於投予時以蒸餾水 將其懸浮而調製。除引起皮膚炎期間爲2 9天之外,其餘 試驗條件均與施實施例1相同。 將皮膚炎症狀數據之演變示於圖5。針對引起皮膚炎 開始第14天(day 14 )之症狀數據,OLL 1 073R-1 + OLS3 05 9 投予群,與 LB MEP 1 705 60 1 + ST MEP 1 70560 1 投予群及未發酵物投予群相比爲低。 於引起皮膚炎開始弟 14天(day 14)之後, OLL 1 073R-1+OLS3 05 9 投予群,與 LB MEP 1 70560 1 + ST MEP 1705601投予群及未發酵物投予群相比,爲有意義的 低。 -27- (24) (24)200826951 由此可知,對異位性皮膚之症狀發揮毫不遜色於本發 明之 OL S3 0 5 9之抑制效果且已於實施例 2中確認之 OLL 1 073R-1,與本發明之Ο L S 3 0 5 9組合使用時,具有抑 制異位性皮膚炎之症狀之效果。 〔實施例4〕 (發酵乳之製造例1 ) 將50.0kg之生乳、5.0kg之脫脂奶粉、8.0kg之砂糖 、1.0kg 之膠原胜肽(商品名:Nippi Peptide,Nippi Collogen化妝品公司製)、0.1kg之含神經醯胺之蒴篛芋 萃取物(商品名:蒴篛神經醯胺,Unitika (股份有限)製 )、0.1kg之香料、33.8kg之水於65°C下混合溶解,製成 調和乳。該調和乳經於65 °C下均質化處理(壓力1〇 〇kg/ cm2)後,於95 °C下進行2分鐘之殺菌處理,再冷卻至43 〜45°C。於該經殺菌處理之調和乳中,添力[I 2.0kg之乳酸 菌菌翫,於塡充至容器後,使其於43 °C下進行4小時之發 酵,而得發酵乳(靜置型優格)。此處乳酸菌菌翫係使用 保加利亞桿菌(Lactobacillus bulgaricus2038,分離自保 加利亞優格LB 8 1 (註冊商標)明治乳業(股份有限)製 ),與嗜熱鏈球菌 OLS3059 ( Streptococcus thermophilus OLS3 05 9 )(以下亦稱做OL S 3 05 9,係分離自保加利亞優 格LB 8 1 (註冊商標)明治乳業股份有限公司製)之等量 培養液。 -28- (25) 200826951 〔實施例5〕 (發酵乳飲料之製造例1 ) 將23.0kg之生乳、12.1kg之脫脂奶粉、62.9kg之水 於70 °C下混合溶解,製成調和乳。該調和乳經於65 °C下 均質化處理(壓力1 50kg/ cm2 )後,於95 °C下進行1〇分 鐘之殺菌處理,再冷卻至40〜45 °C。於該經殺菌處理之調 和乳中,添加2.0kg之乳酸菌菌酝,使其於40〜45 °C下進 ί 行5小時之發酵,而得發酵乳。 此處乳酸菌菌翫係使用保加利亞桿菌(Lactobacillus bulgaricuS2 03 8,分離自保加利亞優格LB81 (註冊商標) 明治乳業(股份有限)製),與嗜熱鏈球菌0LS 3 0 5 9 ( Streptococcus thermophilus OLS3059 ) (以下亦稱做 OLS3 05 9,係分離自保力口利亞優格LB81 (註冊商標)明治 乳業股份有限公司製)之等量培養液。 爲機械性地破壞該發酵乳之凝乳,均質化處j里彳壓方 1 1 50kg/cm2 )後,冷卻至5°C,而得調和發酵乳。另外, 將3.0kg之果膠、2.0kg之砂糖、0.〇2kg之甜味料、〇 lkg 之香料、3 4 · 0 k g之水於6 5 °C下混合溶解,以1 3 〇 ^進行2 秒鐘之殺菌處理後,再冷卻至5 °C而製成糖液。 將調和發酵乳與糖液以規定之濃度混合,而得發酵乳 飮料(攪拌型優格)。 〔實施例6〕 (發酵乳飲料之製造例2 ) -29- (26) 200826951 將23.0kg之生乳、12.1kg之脫脂奶粉、62.9kg之水 於7 0。(:下混合溶解,製成調和乳。該調和乳經於6 5 °C下 均質化處理(壓力150kg/ cm2 )後,於95 °C下進行10分 鐘之殺菌處理,再冷卻至4 0〜4 5 °C。於該經殺菌處理之調 和乳中,添加2.0kg之乳酸菌菌翫,使其於40〜45 °C下進 行5小時之發酵,而得發酵乳。 此處乳酸菌菌翫係使用保加利亞桿菌(Lactobacillus ^ bulgaricus2 03 8,分離自保加利亞優格LB81 (註冊商標) 明治乳業(股份有限)製),與嗜熱鏈球菌OLS 3 05 9 ( Streptococcus thermophilus OLS3059 ) (以下亦稱做 OLS 3 05 9,係分離自保加利亞優格LB81 (註冊商標)明治 乳業股份有限公司製)之等量培養液。 爲機械性地破壞該發酵乳之凝乳,均質化處理(壓力 150kg/ cm2)後,冷卻至5°C,而得調和發酵乳。另外, 將3.0kg之果膠、2.0kg之砂糖、〇.〇2kg之甜味料、i.〇kg " 之膠原胜肽(商品名··膠原胜肽5 2 0 0,新田明膠公司製) 、0.03kg之含神經醯胺之玉米萃取物(商品名:玉米神經 醯胺ME 1-R辻製油(股份有限)製)、0.1kg之香料、 • 3 4 · 0 k g之水於6 5 °C下混合溶解,以1 3 0。〇進行2秒鐘之殺 菌處理後,再冷卻至5 °C而製成糖液。 將調和發酵乳與糖液以規定之濃度混合,而得發酵乳 飮料(攪拌型優格)。 〔實施例7〕 -30- (27) (27)200826951 (乳飲料之製造例) 於lOOOmL之生乳中,添加lg之嗜熱鏈球菌OLS3 05 9 (Streptococcus thermophilus OLS 3 059 )(以下亦稱做 OLS 3 059,係分離自保力口利亞優格LB81 (註冊商標)明治 乳業股份有限公司製)之凍結乾燥粉末,製成調和乳。將 該調和乳均質化處理後,以1 3 0 °C進行3秒鐘之殺菌處理 後,再冷卻至1 〇°C以下。將該經殺菌處理之調和乳塡充於 容器中而得乳飲料。 〔實施例8〕 (根據人體試驗進行皮膚改善效果之驗證) 以前述實施例 4 中之發酵乳 (Streptococcus thermophilus OLS3 05 9,菌體濃度爲 85xl07 ( 10 的 7 次方 )cfu/ g,膠原胜肽爲1000mg/90g,神經醯胺爲300//g /90g)做爲試驗食品,進行攝取量爲90g/次,攝取次數 爲2次/日(早上與晚上各一次),28天之人體實驗。以 3 1位女性(平均年齡爲3 1 · 8歲)爲受試者。 於開始日(攝取試驗食品前)與終止日(第2 8天) ’進行對受試者之皮膚彈性與皮膚紋理密度之評價•觀察 ’及由醫師進行皮疹之觀察。皮膚彈性測定係選擇左或右 邊臉頰,一個位置進行2次測定(商品名:C U Τ Ο Μ E T E R SEM5 7 5,Courage+ Khazaka製)。皮膚紋理密度係選擇 皮膚乾燥部位’測疋1個位置(商品名:V i s u a 1 I m a g e r VI-2 0,Info rward股份有限公司製)。總皮疹數係計算皮 -31 - (28) (28)200826951 疹而進行評價。 於開始日(攝取試驗食品前)與終止日(第28天) ,對受試者本人實施問卷調查。問卷調查之內容示於表1 。如表1所示,針對〔1〕光澤、〔2〕緊緻、〔3〕暗沉 、〔4〕斑點、〔5〕下垂、〔6〕毛孔明顯、〔7〕皮膚紋 理、〔8〕皺紋(額頭)、〔9〕皺紋(嘴角)、〔1 0〕皺 紋(眼角)、〔1 1〕乾燥情形、〔12〕黏腻感、〔1 3〕透 明感、〔1 4〕泛紅、〔1 5〕妝容持久性、〔16〕浮腫、〔 1 7〕便秘、〔1 8〕黑眼圈明顯等1 8個項目,以5個階段 進行評價。再於終止日,受試者本人與醫師進行試驗食品 之有效性評價。 表1 項目 5分 4分 3分 2分 1分 0光澤 有光澤 稍有光澤 難以判定 稍無光澤 無光澤 2)緊緻 有緊緻 稍有緊緻 難以判定 稍無緊緻 不緊緻 3)暗沉 完全不暗沉 幾乎不暗沉 難以判定 稍不暗沉 暗沉 4)斑點 完全無斑點 幾乎無斑點 難以判定 稍無斑點 有斑點 5)下垂 完全無下垂 幾乎無下垂 難以判定 稍無下垂 下垂 6)毛孔明顯 毛孔不明顯 毛孔幾乎不明顯 難以判定 毛孔稍不明顯 毛孔明顯 7)皮膚紋理 紋理細緻 紋理稍細緻 難以判定 紋理稍不細緻 紋理不細緻 8)皺紋(臉部) 完全無皺紋 幾乎無皺紋 難以判定 稍有皴紋 有皺紋 9)皺紋(嘴角) 完全無皺紋 幾乎無皴紋 難以判定 稍有皺紋 有皺紋 10)皺紋(眼周) 完全無皺紋 幾乎無皺紋 難以判定 稍有皺紋 有皺紋 11)乾燥情形 不乾燥 幾乎不乾燥 難以判定 稍乾燥 乾燥 12)黏腻感 沒有黏腻感 幾乎沒有黏腻感 難以判定 稍有黏腻感 有黏腻感 13)透明感 有透明感 稍有透明感 難以判定 稍無透明感 無透明感 η) mi 無泛紅 幾乎無泛紅 難以判定 稍泛紅 泛紅 15)妝容持久性 佳 稍佳 難以判定 稍差 差 16)浮腫 無浮腫 幾乎無浮腫 難以判定 稍浮腫 浮腫 Π)便秘 沒有便秘 幾乎沒有便秘 難以判定 稍有便秘 有便秘 18)黑眼圈 沒有黑眼圈 幾乎沒有黑眼圈 難以判定 稍有黑眼圈 有黑眼圈 -32- (29) (29)200826951 攝取實施例5之試驗食品後,評價皮膚彈性之結果示 於圖6。藉由攝取貫施例5之試驗食品,皮膚彈性有意義 地增加,且改善肌膚光澤。攝取實施例5之試驗食品後, 評價皮膚紋理密度之結果示於圖7。藉由攝取實施例5之 試驗食品,皮膚紋理密度有意義地增加’且改善皮膚紋理 。攝取實施例5之試驗食品後’評價總皮疹數之結果示於 圖8。藉由攝取實施例5之試驗食品,總皮疹數有意義地 減少,且改善肌膚粗糙。 對受試者本人之問卷調查結果示於表2 °確認藉由攝 取試驗食品,對受試者之肌膚狀態及便秘被改善。受試者 本人與醫師進行試驗食品之有效性評價結果示於圖9 °由 受試者本人或醫師確認攝取試驗食品對肌膚有效。5登實嗜 熱鏈球菌 〇LS 3 05 9 ( Streptococcus thermophiUs OLS3 05 9 )具有出色的皮膚改善效果及/或治療效果,確認嗜熱鏈 球菌 OLS 3 05 9 ( Streptococcus thermophilus OLS3 059 )係 最適於使皮膚改善效果及/或治療效果發揮之用途 -33- (30)200826951 暗沉 斑點 下恭 起始日 終止日 起始曰 終止日 起始日 終止曰 起始日 終止日 起始日 終止日 跑始R 饮 f 1- 口 5分(人) 0 0 0 1 0 0 Atk 父口口 於It Η 1 1 0 0 1 1 4分(人) 1 13 2 13 0 7 1 3 1 7 0 4 3分(人) 8 14 6 to 3 14 0 5 3 10 3 7 2分(人) 14 4 16 7 18 14 8 11 19 11 12 14 1分(人) 8 0 7 0 Λ Ο 2 15 I U 11 8 3 15 5 合計(人) 31 31 31 31 31 31 31 31 31 平均分 2.1 3.3 2.1 3.3 1.8 2.8 1.7 2.1 1.9 Ο I 2.7 Ο 1 1.7 31 2.4 數 皮膚‘ 紋理 mm 額頭) 皴紋(嘴角) —雛紋(眼角) 杜fef故 起始曰 終止日 起始曰 終止日 起始日 終止日 起始曰 終止日 起始曰 終止日 ίΡΛΑρ 5分认) 0 0 1 0 0 0 ΰ 終止Η 1 1 0 0 3 1 4分认) 3 10 4 19 iZ 3 8 3 8 4 15 3 11 3分(人) 8 12 7 1Π 6 1U 7 9 5 6 4 3 7 2分认) 12 7 13 η 16 11 11 12 10 12 12 10 1分(人) 8 2 6 2 5 2 11 5 11 0 10 2 合計认) 31 31 31 31 31 31 31 31 31 ai 平均分 2.2 3.0 2.4 3.0 2..3 2.8 2.0 2.6 2.1 J 1 3.1 31 2.3 31 3.0 數The antibody manufactured by Pharmingen Co., Ltd. was measured by changing one of the recommended samples of the company. The dust-crushing liquid was prepared in the following manner. Adults of J Dermatophagoides pterony s sinus (manufactured by Mite-Dp Co., Ltd.) were degreased with anhydrous methanol, and distilled water was added for ultra-crushing treatment. Next, the water-soluble fractions were separated by centrifugation, and the knot was dried, and the protein concentration was adjusted to 4.5 mg/ml with distilled water; BSA conversion, and the DC protein assay of Bio RAD was used to determine the dermatitis test system. The method of Habitat et al. (Hai Yezhe - Other, Allergy 5 0 : 1 1 5 2 - 1 1 6 2, 2 0 0 1 ) was carried out. The sputum was used to cause the atopic dermatitis site (head, outer ear and neck) and the SDS solution (4%) was applied to all the sites causing the ectopic dermatitis 20 times. The coating amount is about 20 0 / 1 for a total of 20 times. After the S D S grass squid was applied, the dermatitis test group was coated with dust _ breakage liquid, and the non-anti-inflammatory test group was coated with distilled water, and the same was applied 20 times. Borrow. The evolution of each 7.13 10 is taken from the blood. 验 Use the step-by-step test! 冻Lake the LSL sound wave (to be determined) History and knife shaving, the peeling of the skin solution is repeated by the above steps every -23- (20) 200826951 B Causes dermatitis, repeated 7 days after the start of the test (day 〇), until the end of the test. IgE was measured in the following manner. That is, an anti-IgE antibody (manufactured by Pharmingen Co., Ltd.) of 2 // g/ml -1 antibody was added to a 96-well plate, and it was allowed to stand at 37. . Next 1 hour. After washing with Tween 20/PBS (PBS-Tween, 0.05%), BS A/PBS (1%) was added and allowed to stand at room temperature for 30 minutes. After washing with PBS-Tween, a serum sample was added and a standard {curve IgE (manufactured by Pharmingen Co., Ltd.) was prepared and left to stand at room temperature for 30 minutes. After washing with PBS-Tween, biotin-anti-IgE antibody (manufactured by Pharmingen Co., Ltd.) was added to the antibody twice at 0.5//g/ml, and the mixture was allowed to stand at room temperature for 1 hour. After washing with PBS-Tween, Streptavidin-HRP (manufactured by Pharmingen Co., Ltd.) was added, and the mixture was allowed to stand at room temperature for 30 minutes. After washing with PBS-Tween, D TMB + Substrate Chromogen (manufactured by DAKO Co., Ltd.) was added, and a color reaction was started. Sulfuric acid (1N) was added after the color reaction, and the absorbance (450 nm) was measured by a microplate reader, and the IgE concentration in the serum was calculated. The evolution of dermatitis-like data is shown in Figure 1. The data system is based on [1 • 〕 formation of dry skin • suede, [2] redness and bleeding, [3] tissue shedding · abrasion, [4] edema, [5] itching behavior and other five items, with "0 · Asymptomatic, "1. Mild", "2. Moderate", "3. Severe" and other four stages, calculate and evaluate each data obtained by each mouse. The symptom data for the 13th day (day 13) at the start of dermatitis was administered as the LS3 05 9 group was significantly lower than the distilled water group. Conversely, there was no significant difference between the ATCC 53 1 03 group and the distilled water group. -24- (21) (21) 200826951 The total IgE concentration is shown in Fig. 2. The concentration of IgE administered to the OLS3059 group was significantly lower than that of the distilled water group for the total IgE in the serum on the 15th day after the onset of dermatitis (day 15). Conversely, the concentration of IgE was slightly lower in the ATCC 5 3 1 03 group than in the distilled water group, but there was no statistical difference. It is known that ATCC 5 3 1 03 is a lactic acid bacterium having a therapeutic effect on atopic skin, but OLS 3 05 9 has a higher inhibitory effect on symptoms than ectopic skin than ATCC 5 3 1 0 3 . [Example 2] (Verification of the inhibitory effect of atopic dermatitis according to the in vivo test) The female 4-week-old NC/Nga mouse (produced by Japan SLC (stock limited)) was domesticated and reared as follows. group. The steamed water group (n=4) was administered to a population of Bulgarian bacillus OLL1073R-1 (Lactobacillus delbrueckii subspecies bulgaricus OLL1073R-1, hereinafter also referred to as OLL 1 073 R-1) (n=6). Streptococcus thermophilus OLS 3 05 9 (Streptococcus thermophilus OLS 3 05 9, isolated from the Bulgarian Yogurt LB81 (made by Meiji Dairy Co., Ltd.) group (η = 5). These mice were given free feed. (trade name: CRF_1, Oriental Yeast Industry Co., Ltd.) and steaming water. The other test conditions were the same as in Example 1 except for 30 days during dermatitis. Shown in Figure 3. For the symptom data of the 13th day (day 13) that caused dermatitis-25-(22) (22)200826951, the OLL 1 073 R-1 group was administered and administered to the OLS3 05 9 group and administered. The distilled water group was considered to be significantly lower. The total IgE concentration is shown in Figure 4. For the total IgE in serum, the OLL 1 073 R-1 population was administered compared to the OLS 3 0 5 9 group and the distilled water group was administered. The IgE concentration was considered to be low. It is known that the OLS3 059 of the present invention has an inhibitory effect on the symptoms of atopic dermatitis. In addition, OLL 1 073R-1 is also found to have an inhibitory effect on the symptoms of atopic dermatitis, but the IgE concentration system. The OLS3059 of the present invention is lower. [Example 3] (according to in vivo Verification of the inhibitory effect of atopic dermatitis. For the symptoms of atopic dermatitis, the OLL 1 073 R- which is inferior to the OLS3 05 9 of the present invention and which has been confirmed in Example 2 will be exerted. 1. In combination with the 0 LS 3 0 5 9 of the present invention, the symptoms of atopic dermatitis were examined. Two types of Lactobacillus delbrueckii subspecies isolated from the Bulgarian Yogur (Meiji Dairy Co., Ltd.) were used. Bulgaricus MEP1705601, in combination with Streptococcus thermophilus MEP 1 705 60 1 , is a comparison object of the combination of OLS 3 05 9 and OLL 1 07 3R-1 of the present invention. Female 4-week NC/Nga mice (Japan SLC (limited stock) )), after domestication, the following four groups are grouped: Lactobacillus delbrueckii subspecies bulgaricus OLL1073R-1, and Streptococcus thermophilus -26- (23) (23) 200826951 OLS 3 05 9 (The following is also referred to as 〇LL1〇73R] + OLS3 05 9 to the group) (n= 8). Lactobacillus delbrueckii subspecies bulgaricus MEP 1 705 60 1 ' is a group of fermented products prepared with Streptococcus thermophilus MEP 1 705 60 1 (also known as lb MEP 1705601 + STMEP 1705601 administered to the group) (n=9). A group of unfermented foods (n=9), and a group of non-derived auxin and un-fermented (hereinafter also referred to as non-inducing group) (n 2 4). The intake of fermented materials and unfermented materials was 100 mg / mouse / day, and the daily oral administration was continued until the end of the test. The number of bacteria in the fermented product was LXL 1 073 R-1 was lxl 〇 9 cfu / g, OLS3 05 9 was 3xl 09 9 cfu / g, LB MEP 1 70560 1 was 8.6 χ 108 cfu / g, and ST ΜΕΡ 1705601 was 2.1 x l 〇9cfu/g. The fermented product and the unfermented product were freeze-dried, and then suspended in distilled water at the time of administration to prepare. The remaining test conditions were the same as in Example 1 except that the period of dermatitis was 29 days. The evolution of dermatitis-like data is shown in Figure 5. For the symptom data of the 14th day (day 14) that caused dermatitis, OLL 1 073R-1 + OLS3 05 9 was administered to the group, and LB MEP 1 705 60 1 + ST MEP 1 70560 1 was administered to the group and unfermented product. Compared to the group is low. After 14 days (day 14) of dermatitis, OLL 1 073R-1+OLS3 05 9 was administered to the group, compared with LB MEP 1 70560 1 + ST MEP 1705601 administered group and unfermented product group. It is meaningful low. -27- (24) (24) 200826951 It can be seen that the symptoms of atopic skin exert an inhibitory effect of OL S3 0 5 9 which is inferior to the present invention and which has been confirmed in Example 2, OLL 1 073R- 1. When used in combination with Ο LS 3 0 5 9 of the present invention, it has an effect of suppressing the symptoms of atopic dermatitis. [Example 4] (Production Example 1 of fermented milk) 50.0 kg of raw milk, 5.0 kg of skim milk powder, 8.0 kg of granulated sugar, 1.0 kg of collagen peptide (trade name: Nippi Peptide, manufactured by Nippi Collogen Cosmetics Co., Ltd.), 0.1 kg of a cerium containing neuropterin extract (trade name: sputum ceramide, manufactured by Unitika (limited by shares)), 0.1 kg of perfume, and 33.8 kg of water are mixed and dissolved at 65 ° C. Blend milk. The blended milk was homogenized at 65 ° C (pressure 1 〇 〇 kg / cm 2 ), then sterilized at 95 ° C for 2 minutes, and then cooled to 43 ~ 45 ° C. In the sterilized treated milk, I added a force of [I 2.0 kg of lactic acid bacteria, and after fermenting it to a container, it was fermented at 43 ° C for 4 hours to obtain fermented milk (stationary type of yogurt) ). Here, the lactic acid bacteria strain uses Lactobacillus bulgaricus 2038 (isolated from Bulgarian Yogurt LB 8 1 (registered trademark) Meiji Dairy (stock limited)), and Streptococcus thermophilus OLS3059 (Streptococcus thermophilus OLS3 05 9 ) (hereinafter also It is called OL S 3 05 9, and is an equal amount of the culture liquid separated from the Bulgarian Yogurt LB 8 1 (registered trademark) Meiji Dairy Co., Ltd.). -28- (25) 200826951 [Example 5] (Production Example 1 of fermented milk beverage) 23.0 kg of raw milk, 12.1 kg of skim milk powder, and 62.9 kg of water were mixed and dissolved at 70 ° C to prepare a blended milk. The blended milk was homogenized at 65 °C (pressure: 50 kg/cm2), then sterilized at 95 °C for 1 Torr, and then cooled to 40 to 45 °C. To the sterilized treated milk, 2.0 kg of lactic acid bacteria mash was added, and the fermentation was carried out at 40 to 45 ° C for 5 hours to obtain fermented milk. Here, Lactobacillus bulgaricu S2 03 8, isolated from Bulgarian Yogurt LB81 (registered trademark) Meiji Dairy Co., Ltd., and Streptococcus thermophilus OLS3059 (Streptococcus thermophilus OLS3059) The following is also referred to as OLS3 05 9, which is an equal amount of the culture solution separated from the LV81 (registered trademark) Meiji Dairy Co., Ltd.). In order to mechanically destroy the curd of the fermented milk, the homogenization point is 1 1 50 kg/cm 2 ), and then cooled to 5 ° C to obtain a blended fermented milk. In addition, 3.0 kg of pectin, 2.0 kg of granulated sugar, 0. 〇 2 kg of sweetener, 〇lkg of spices, 3 4 · 0 kg of water were mixed and dissolved at 65 ° C, and carried out at 1 3 〇 ^ After 2 seconds of sterilization, it was cooled to 5 °C to make a sugar solution. The fermented milk and the sugar liquid are mixed at a prescribed concentration to obtain a fermented milk material (stirred type). [Example 6] (Production Example 2 of fermented milk beverage) -29- (26) 200826951 23.0 kg of raw milk, 12.1 kg of skimmed milk powder, and 62.9 kg of water were 70%. (: Dissolve and dissolve to prepare a blended milk. The blended milk is homogenized at 65 ° C (pressure 150kg / cm2), then sterilized at 95 °C for 10 minutes, and then cooled to 4 0~ 4 5 ° C. In the sterilized treated milk, 2.0 kg of lactic acid bacteria mash was added and fermented at 40 to 45 ° C for 5 hours to obtain fermented milk. Bulgarian bacillus (Lactobacillus ^ bulgaricus 2 03 8, isolated from Bulgarian Yogurt LB81 (registered trademark) Meiji Dairy (stock limited)), and Streptococcus thermophilus OLS 3 05 9 (Streptococcus thermophilus OLS3059) (hereinafter also referred to as OLS 3 05 9, is an equal amount of culture liquid separated from the Bulgarian Yogurt LB81 (registered trademark) Meiji Dairy Co., Ltd.). The mammary milk of the fermented milk is mechanically destroyed, homogenized (pressure 150kg/cm2), and then cooled. The fermented milk is blended to 5 ° C. In addition, 3.0 kg of pectin, 2.0 kg of granulated sugar, 〇. 〇 2 kg of sweetener, i. 〇 kg " collagen peptide (trade name · collagen) Peptide 5 2 0 0, manufactured by Xintian Gelatin Co., Ltd.) 0.03kg of corn sulphate-containing corn extract (trade name: corn ceramide ME 1-R 辻 oil (limited stock)), 0.1kg of spices, • 3 4 · 0 kg of water at 65 ° C After mixing and dissolving, it is sterilized by 1300 ° for 2 seconds, and then cooled to 5 ° C to prepare a sugar liquid. The fermented milk is mixed with the sugar liquid at a predetermined concentration to obtain a fermented milk mash ( Stirring type yoghurt. [Example 7] -30- (27) (27) 200826951 (Production example of milk beverage) Add lg of Streptococcus thermophilus OLS3 05 9 (Streptococcus thermophilus OLS 3 059) in raw milk of 1000 mL (hereinafter also referred to as OLS 3 059, which is a freeze-dried powder which is separated from the LV81 (registered trademark) Meiji Dairy Co., Ltd.), and is made into a blended milk. After homogenizing the blended milk After sterilizing treatment at 1300 ° C for 3 seconds, the mixture was cooled to 1 〇 ° C or lower. The sterilized mash was filled in a container to obtain a milk beverage. [Example 8] (According to Verification of skin improvement effect by human test) Fermented milk in the aforementioned Example 4 (St Reptococcus thermophilus OLS3 05 9, the bacterial concentration of 85xl07 (10 7th power) cfu / g, collagen peptide 1000mg / 90g, neural guanamine 300 / / g / 90g) as a test food, the intake is 90 g / time, the number of ingestions was 2 times / day (every morning and evening), 28 days of human experiment. A total of 31 women (mean age 3 1 · 8 years old) were enrolled. The skin elasticity and skin texture density of the subject were evaluated on the start date (before ingestion of the test food) and the termination date (the 28th day). Observations were made and the rash was observed by the physician. The skin elasticity measurement system selects the left or right cheek, and the measurement is performed twice in one position (trade name: C U Τ Ο Μ E T E R SEM5 7 5, manufactured by Courage+ Khazaka). The skin texture density was selected from the skin dry area by one position (trade name: V i s u a 1 I m a g e r VI-2 0, manufactured by Inforward Co., Ltd.). The total rash number was calculated by calculating the skin -31 - (28) (28)200826951 rash. On the start date (before ingesting the test food) and the termination date (day 28), the subject was asked to conduct a questionnaire survey. The contents of the questionnaire are shown in Table 1. As shown in Table 1, for [1] gloss, [2] firmness, [3] dullness, [4] spots, [5] sagging, [6] obvious pores, [7] skin texture, [8] wrinkles (forehead), [9] wrinkles (mouth corner), [1 0] wrinkles (eye corners), [1 1] dryness, [12] sticky feeling, [1 3] transparency, [14] redness, 1 5] Long-lasting makeup, [16] edema, [17] constipation, [18] dark circles, and other 18 items were evaluated in 5 stages. On the termination date, the subject and the physician performed an evaluation of the effectiveness of the test food. Table 1 Item 5 points 4 points 3 points 2 points 1 point 0 Glossy shiny slightly shiny is difficult to judge slightly dull and dull 2) Firming has a firmness and a little firmness is difficult to determine a little tight and not tight 3) Dark Sinking is not dull, it is almost dull, it is difficult to judge that it is not dull and dull. 4) The spot is completely spotless, almost no spots, it is difficult to determine that there are no spots and spots. 5) The sagging is completely sagging, and there is no drooping. It is difficult to determine that there is no sagging droop 6) The pores are obviously small, the pores are not obvious, and the pores are hardly obvious. It is difficult to determine that the pores are not obvious. The pores are slightly inconspicuous. 7) The texture of the skin is fine and the texture is slightly fine. It is difficult to judge the texture. The texture is not fine. The texture is not fine. 8) Wrinkles (face) It is difficult to judge wrinkles without wrinkles. Slightly wrinkles and wrinkles 9) Wrinkles (mouth corners) Completely wrinkle-free, almost flawless, difficult to determine, slightly wrinkled and wrinkled 10) Wrinkles (eyes around) Completely wrinkle-free, almost wrinkle-free, difficult to determine, slightly wrinkled and wrinkled 11) Dry condition It is difficult to judge that it is slightly dry and dry without drying. 12) Stickiness is not sticky. There is almost no sticky feeling. There is a sticky feeling and a sticky feeling. 13) Transparency has a transparent feeling. It is slightly transparent. It is difficult to judge. There is no transparency. No transparency. η) mi No redness, almost no redness, no difficulty, no redness, no redness. 15) Makeup persistence Good slightly better difficult to determine a little difference 16) edema without edema almost no edema difficult to determine a slight edema edema) constipation no constipation almost no constipation difficult to determine a little constipation constipation 18) dark circles no dark circles almost no dark circles difficult to determine slightly Dark circles with dark circles - 32 - (29) (29) 200826951 After ingesting the test food of Example 5, the results of evaluating skin elasticity are shown in Fig. 6. By ingesting the test food of Example 5, the skin elasticity is meaningfully increased and the skin's gloss is improved. The results of evaluating the skin texture density after ingesting the test food of Example 5 are shown in Fig. 7. By ingesting the test food of Example 5, the skin texture density was significantly increased' and the skin texture was improved. The results of evaluating the total number of rashes after ingesting the test food of Example 5 are shown in Fig. 8. By ingesting the test food of Example 5, the total number of rashes was significantly reduced, and the skin roughness was improved. The results of the questionnaire survey of the subjects themselves are shown in Table 2 °. It was confirmed that the skin condition and constipation of the subjects were improved by taking the test foods. Subjects The results of the evaluation of the effectiveness of the test foods performed by the physician and the physician are shown in Fig. 9 °. It is confirmed by the subject or the physician that the ingestion of the test food is effective for the skin. 5 Streptomyces faecalis LS 3 05 9 (Streptococcus thermophiUs OLS3 05 9 ) has excellent skin improvement and/or therapeutic effect, confirming that Streptococcus thermophilus OLS 3 05 9 (Streptococcus thermophilus OLS3 059 ) is most suitable for making Use of skin improvement and/or therapeutic effect-33- (30)200826951 Under the dark spot, the start date of the start date, the start date of the end date, the start date of the start date, the start date of the start date of the start date of the start date R drink f 1-port 5 points (person) 0 0 0 1 0 0 Atk father mouth in It Η 1 1 0 0 1 1 4 points (person) 1 13 2 13 0 7 1 3 1 7 0 4 3 points ( Person) 8 14 6 to 3 14 0 5 3 10 3 7 2 points (person) 14 4 16 7 18 14 8 11 19 11 12 14 1 point (person) 8 0 7 0 Λ Ο 2 15 IU 11 8 3 15 5 Total (person) 31 31 31 31 31 31 31 31 31 Average score 2.1 3.3 2.1 3.3 1.8 2.8 1.7 2.1 1.9 Ο I 2.7 Ο 1 1.7 31 2.4 Number of skin 'texture mm forehead 皴 pattern (mouth angle) - crepe (eye corner) Du fef, the beginning of the start date, the start date, the end of the start date Day start 曰 end day start 曰 end day ΡΛΑ 5 5 5 points) 0 0 1 0 0 0 ΰ End time 1 1 0 0 3 1 4 points) 3 10 4 19 iZ 3 8 3 8 4 15 3 11 3 points (人) 8 12 7 1Π 6 1U 7 9 5 6 4 3 7 2 points) 12 7 13 η 16 11 11 12 10 12 12 10 1 point (person) 8 2 6 2 5 2 11 5 11 0 10 2 Total Recognition 31 31 31 31 31 31 31 31 31 ai Average score 2.2 3.0 2.4 3.0 2..3 2.8 2.0 2.6 2.1 J 1 3.1 31 2.3 31 3.0
-34- (31) (31)200826951 〔實施例9〕 (根據人體試驗進行整腸改善效果與皮膚改善效果之關係 之驗證) 以則述貫施例 5 中之發酵乳(Streptococcus thermophilus OLS3 05 9,菌體濃度爲 50xl07 ( 1〇 的 7 次方 )cfu/ g ),及實施例 6中之發酵乳(Streptococcus thermophilus OLS3 0 5 9,菌體濃度爲 50xl07 ( 1〇 的 7 次方 )cfu/g,膠原胜肽爲 l〇〇〇mg/l20ml,神經醯胺爲 300 //g/120ml)做爲試驗食品,進行攝取量爲120ml /次 ,攝取次數爲2次/日(早上與晚上各一次),28天之人 體實驗。實施例5中之發酵乳飮料以2 8位女性(平均年 齢爲3 1 . 1歲),實施例6中之發酵乳飮料,以2 8位女性 (平均年齡爲30.3歲)爲受試者。 與實施例8相同,對受試者本人實施問卷調查。問卷 調查之內容如表1所示。對受試者本人問卷調查之結果示 於表3及表4。確認藉由攝取試驗食品,對受試者之肌膚 狀態及便秘被改善。針對表1之〔1〕光澤、〔2〕緊緻、 〔7〕皮膚紋理、〔i i〕乾燥情形、〔1 5〕妝容持久性、 〔1 7〕便秘等6個項目,於實施例6之試驗食品(含膠原 胜肽及神經醯胺),具有較實施例5之試驗食品優異之效 果。 -35- (32) 200826951 表 ί i 膠原蛋白 f澤 % 1緻 01 3ΪΚ 斑點 ^ J 下垂 主刀卩Η甜 起始曰 終止曰 起始日 終止日 起始曰 終止曰 起始曰 終止日 起始曰 終止日 紀始R iA rU □ 5分(人) 0 0 0 0 0 0 0 於止b 2 0 0 1 0 4分(人) 2 9 3 9 2 7 5 6 3 7 3 4 3分(人) 7 10 11 12 4 7 3 3 11 10 2 8 2分(人) 17 8 10 7 15 12 11 11 11 8 14 15 1分(人) 2 1 4 0 7 2 8 6 3 3 8 1 合計(人) 28 28 28 28 28 28 28 28 28 28 28 平均分數 2.3 3.0 2.5 3.1 2.0 ί 2.7 2.3 2.5 2.5 2.8 2 1 28 25 j 皮膚 紋理 皺紋C 額頭) 皺紋(嘴角) 皺紋(眼角) 起始曰 終止日 起始曰 終止曰 起始日 終止日 起始曰 終止日 起始日 糸冬±日 起始日 5分(人) 2 1 2 2 2 2 0 本公ill ϋ 1 0 0 0 2 4分(人) 2 6 5 9 g 12 3 7 0 4 8 12 3分(人) 6 9 6 3 5 8 4 2 2 3 8 10 2分(人) 16 9 11 11 13 6 18 13 6 11 9 3 1分(人) 2 3 4 3 0 0 5 3 20 10 3 1 合計(人) 28 28 28 28 28 28 28 28 28 28 no TO 平均分數 2.5 2.8 2.6 2.9 3.0 3.4 2.1 2.6 1.4 2.0 Zo 2.8 Zo 3.4 5分(人) 4分(人) 3分(人) 2分(人) 1分(人) 合計(人) 平均分數 透弓 月感 泛 .紅 妝容持久性 浮腫 便秘 里眼囫日日願 起始日 終止曰 起始日 終止日 起始曰 終止曰 起始曰 終止曰 起始曰 終止日 趣台日 終止日 0 0 2 1 2 2 1 4 0 2 0 2 2 4 7 7 2 11 6 10 0 9 2 6 3 13 2 5 5 8 3 6 0 4 6 3 20 10 13 14 17 7 14 6 10 10 5 7 3 1 4 1 2 0 4 2 18 3 15 10 28 28 28 28 28 28 28 28 28 28 28 28 2.1 2.7 2.6 2.8 2.5 3.3 2.5 3.3 1.4 2.9 1.8 2.4 將整腸(通便)數據之演變示於表4。數據化係以 -36- (33) 200826951 1 .無便秘」、「2 .幾乎無便秘」、「3 .無法判定」、「4 . 稍有便秘」、「5.有便秘」5階段進行,算出各受試者攝 取實驗食品前及攝取後之差,並加以評價。例如攝取實驗 食品前爲「5 .有便秘」之受試者,於攝取後變爲「3 .無法 判定」時,解析爲「整腸改善爲「2以上」」。 -37- (34)200826951 ί澤 % m 01 m -4 1T 社 .斑點 肢· 1 有 下垚 J 右21 口口甜 起始曰 終止日 起始曰 終止日 起始曰 終止日 起始曰 終止曰 起始曰 終止日 5分(人) 0 0 1 0 0 0 也夂口口 衫itu 1 2 1 1 0 2 4分(人) 2 10 4 17 1 8 1 3 6 10 5 8 3分(人) 9 14 8 7 1 1 1 11 2 7 5 7 6 3 2分(人) 11 2 9 3 21 8 17 13 11 9 13 14 1分(人) 6 2 8 1 5 1 1 7 3 5 1 4 1 合計(人) 28 28 28 28 28 28 28 28 28 28 平均分數 2..3 3-1 1 2.5 3.4 1.9 | 2.9 2.0 2.6 2.5 3.0 Zo 2.4 28 2.9 膠原蛋_ •神經醯胺· 「有 皮膚 麵 皺紋( 額頭) 駿紋(嘴角) 激紋(眼角) 乾煶愔形 起始日 終止日 起始日 終止日 起始日 終止日 起始曰 終止日 起始曰 終止日 紀始闩 软丨{- p 5分(人) 0 0 0 2 1 1 於It㈡ 2 2 0 0 3 3 4分(人) 5 9 10 9 Q 6 12 〇 15 0 13 11 15 3分(人) 8 14 4 5 J 3 3 5 0 2 6 5 2分(人) 11 4 9 9 1 λ 12 8 1U 8 10 10 8 5 1分(人) 4 1 5 3 4 1 5 1 18 3 0 0 合計(人) 28 28 28 28 28 28 28 28 28 28 28 平均分數 2.5 3.1 2.7 2.9 2.7 3.3 2.6 3.2 1.4 2.9 3.3 Z5 3.6 透弓 臓 泛紅 妝容持久性 浮腫 便秘 里阻願昍顥 起始曰 終止日 起始曰 終止日 起始曰 終止日 起始曰 終止日 起始日 終止日 ^^0 終止日 5分(人) 0 1 2 2 0 2 0 3 0 7 1 1 4分(人) 2 5 11 10 5 16 11 15 0 5 3 6 3分(人) 8 15 4 3 6 8 1 3 1 5 2 6 2分(人) 14 6 7 11 15 2 15 5 17 8 12 13 1分(人) 4 1 4 2 L 0 1 2 10 3 10 2 合計(人) 28 28 28 28 28 28 28 28 28 28 28 28 平均分數 2.3 3.0 3.0 3.0 2.5 3.6 2.8 3.4 1.7 3.2 2.0 2.7 攝取實施例5及6之試驗食品後,評價皮膚彈性之結 -38- (35) 200826951 果示於圖10〜12。圖1〇係「整腸改善爲「1以上」」之 受試者,圖11係「整腸改善爲「2以上」」之受試者,圖 1 2係「整腸改善爲「3以上」」之受試者評價皮膚彈性後 . 之結果。證實對整腸(通便)改善效果較高者,皮膚改善 效果亦高。該傾向於實施例6之試驗食品(含膠原胜肽及 神經醯胺),具有較實施例5之試驗食品優異之效果。藉 由攝取實施例5及6之試驗食品,可增加肌膚彈性,改善 ( m誠、澤。 攝取實施例5及6之試驗食品後,評價皮膚紋理密度 之結果不於圖13〜15。圖13係「整腸改善爲「1以上」 」之受試者,圖1 4係「整腸改善爲「2以上」」之受試者 ’圖1 5係「整腸改善爲「3以上」」之受試者評價皮膚紋 理密度後之結果。證實對整腸(通便)改善效果較高者, 皮膚改善效果亦高。該傾向於實施例4之試驗食品(含膠 原胜肽及神經醯胺),具有較實施例5之試驗食品優異之 (效果。藉由攝取實施例5及6之試驗食品,可增加皮膚紋 理密度,改善皮膚紋理。 • 根據本發明可提供因來自天然物質而具高安全性’進 - 而其皮膚改善效果及/或治療效果高,且含有製造特性優 良的乳酸菌之組成物以及發酵乳。進而可提供特徵爲對整 腸改善效果及/或治療效果較高者,皮膚改善效果及/或 治療效果亦高之發酵乳。 【圖式簡單說明】 -39- (36) (36)200826951 〔圖 1〕於蒸飽水投予群,Lactobacillus rhamnosus ATCC 5 3 1 0 3 投予群,Streptococcus thermophilus OLS 3 05 9投予群,及非引發皮膚炎群,試驗期間中之皮膚 炎症狀數據之演變圖(以平均値i:標準偏差値表示。 **Ρ<0·01,*Ρ<0·05,Dunnett 之多重比較)。 〔圖 2〕於蒸餾水投予群,Lactobacillus rhamnosus ATCC 5 3 1 0 3 投予群,Streptococcus thermophilus OLS 3 05 9投予群,及非引發皮膚炎群,15天後(day 15 ) 之血清中總IgE濃度圖(以平均値±標準偏差値表示。 * * P < 0.0 1 5 Dunnett 之多重比較)。 〔圖 3〕於蒸 f留水投予群,Lactobacillus bulgaricus OLL1073R-1 投予群,及 Streptococcus thermophilus OLS 3 05 9投予群,試驗期間中之皮膚炎症狀數據之演變圖 (以平均値土標準偏差値表示。**Ρ<0.01,*Ρ<〇·〇5, Dunnett之多重比較)。 〔圖 4〕於蒸餾水投予群,Lactobacillus bulgaricus OLL1073R-1 投予群,及 Streptococcus thermophilus OLS3 05 9投予群,15天後(day 15 )之血清中總IgE濃度 圖(以平均値土標準偏差値表示。**Ρ<〇·〇1,Dunnett之多 重比較)。 〔圖 5〕方令以 Lactobacillus bulgaricus OLL1073R-1 與 Streptococcus thermophilus OLS3059 所調製之發酵物 之投予群,及以 Lactobacillus delbrueckii subspecies bulgaricus MEP 1 70560 1 ,與 Streptococcus thermophilus -40- (37) (37)200826951 MEP1705601所調製之發酵物之投予群,未發酵物投予群 及非引發皮膚炎且未發酵物投予群,試驗期間中之皮膚炎 症狀數據之演變圖(以平均値土標準偏差値表示。 **P<0.01,*P<0.05,Dunnett 之多重比較)。 〔圖 6〕以嗜熱鍵球菌 OLS3059 ( Streptococcus thermophilus OLS3 05 9 )所調製之發酵乳做爲試驗食品, 受試者皮膚彈性圖。 〔圖 7〕以嗜熱鏈球菌 OLS 3 0 59 ( Streptococcus thermophilus OLS 3 05 9 )所調製之發酵乳做爲試驗食品, 受試者皮膚紋理密度圖。 〔圖 8〕以嗜熱鍵球菌 OLS3059 ( Streptococcus thermophilus OLS3059)所調製之發酵乳做爲試驗食品, 受試者總皮疹數之觀察結果圖。 〔圖 9〕以嗜熱鏈球菌 OLS3059 ( Streptococcus thermophilus OLS 3 05 9 )所調製之發酵乳做爲試驗食品, 根據受試者及醫師進行有效性評價結果圖。 〔圖 10〕以嗜熱鏈球菌 OLS3059 ( Streptococcus thermophilus OLS 3 05 9 )所調製之發酵乳做爲試驗食品, 「整腸改善爲「1以上」」之受試者其皮膚彈性圖。 〔圖 11〕以嗜熱鏈球菌 OLS 3 059 ( StrePtococcus thermophilus OLS3059)所調製之發酵乳做爲§式驗食品’ 「整腸改善爲「2以上」」之受試者其皮膚彈性圖。 〔圖 12〕以嗜熱鏈球菌 OLS3 059 ( Streptococcus thermophilus OLS 3 059 )所調製之發酵乳做爲試驗食品’ -41 - (38) 200826951 「整腸改善爲「3以上」」之受試者其皮膚 〔圖13〕以嗜熱鏈球菌OLS3 05 9 thermophilus OLS3059)所調製之發酵乳估 「整腸改善爲「1以上」」之受試者其皮膚 〔圖14〕以嗜熱鏈球菌 OLS3 05 9 thermophilus OLS 3 05 9 )所調製之發酵乳估 「整腸改善爲「2以上」」之受試者其皮膚 〔圖 15〕以嗜熱鏈球菌 OLS3 05 9 thermophilus OLS3059)所調製之發酵乳倘 「整腸改善爲^ 3以上」」之受試者其皮膚 彈性圖。 〔Streptococcus :爲試驗食品, 紋理密度圖。 〔Streptococcus :爲試驗食品, 紋理密度圖。 〔Streptococcus :爲試驗食品’ 紋理密度圖。 -42--34- (31) (31) 200826951 [Example 9] (Verification of the relationship between the effect of improving the intestines and the skin improving effect according to the human test) The fermented milk of the Example 5 (Streptococcus thermophilus OLS3 05 9) , the cell concentration is 50xl07 (1 〇 7th power) cfu / g), and the fermented milk of Example 6 (Streptococcus thermophilus OLS3 0 5 9, the cell concentration is 50xl07 (1 〇 7th power) cfu / g, collagen peptide is l〇〇〇mg/l20ml, neural guanamine is 300 //g/120ml) as test food, the intake is 120ml / time, the intake frequency is 2 times / day (in the morning and evening Once), 28 days of human experiment. The fermented milk mash in Example 5 was made up of 28 females (average age 3 31. 1 year old), fermented milk mash in Example 6, and 28 females (average age 30.3 years old) as subjects. In the same manner as in Example 8, a questionnaire survey was conducted on the subject himself. The contents of the questionnaire are shown in Table 1. The results of the subject's own questionnaire are shown in Tables 3 and 4. It was confirmed that the skin condition and constipation of the subject were improved by ingesting the test food. For the six items of [1] gloss, [2] firming, [7] skin texture, [ii] dry condition, [15] makeup persistence, [17] constipation, etc. in Table 1, The test food (containing collagen peptide and neuropterin) had an excellent effect compared to the test food of Example 5. -35- (32) 200826951 Table ί i Collagen f Ze % 1 to 01 3 ΪΚ Spot ^ J Drooping main knife 卩Η sweet start 曰 termination 曰 start date end date start 曰 start 曰 start 曰 end day start 曰End of day R iA rU □ 5 points (person) 0 0 0 0 0 0 0 at stop b 2 0 0 1 0 4 points (person) 2 9 3 9 2 7 5 6 3 7 3 4 3 points (person) 7 10 11 12 4 7 3 3 11 10 2 8 2 points (person) 17 8 10 7 15 12 11 11 11 8 14 15 1 point (person) 2 1 4 0 7 2 8 6 3 3 8 1 Total (person) 28 28 28 28 28 28 28 28 28 28 28 Average score 2.3 3.0 2.5 3.1 2.0 ί 2.3 2.7 2.3 2.5 2.8 2 1 28 25 j Skin texture wrinkles C Forehead) Wrinkles (mouth corner) Wrinkles (eye corners) Start 曰 End of day曰 termination 曰 start date start date start date start date start winter ± day start date 5 minutes (person) 2 1 2 2 2 2 0 gong ill ϋ 1 0 0 0 2 4 points (person) 2 6 5 9 g 12 3 7 0 4 8 12 3 points (person) 6 9 6 3 5 8 4 2 2 3 8 10 2 points (person) 16 9 11 11 13 6 18 13 6 11 9 3 1 point (person) 2 3 4 3 0 0 5 3 20 10 3 1 Total (person) 28 28 28 28 28 28 28 28 28 28 no TO Average score 2.5 2.8 2.6 2.9 3.0 3.4 2.1 2.6 1.4 2.0 Zo 2.8 Zo 3.4 5 points (person) 4 points (person) 3 points (person) 2 points (person) 1 point (person) Total (person) Average score through the bow of the moon. Red makeup lasting edema constipation in the eyelids day to start Day Termination 曰 Start Date End Date Start End 曰 Start End 曰 Start 曰 End Day End Date 0 0 2 1 2 2 1 4 0 2 0 2 2 4 7 7 2 11 6 10 0 9 2 6 3 13 2 5 5 8 3 6 0 4 6 3 20 10 13 14 17 7 14 6 10 10 5 7 3 1 4 1 2 0 4 2 18 3 15 10 28 28 28 28 28 28 28 28 28 28 28 28 2.1 2.7 2.6 2.8 2.5 3.3 2.5 3.3 1.4 2.9 1.8 2.4 The evolution of the intestine (defecation) data is shown in Table 4. The data is based on -36- (33) 200826951 1 . No constipation, 2 . Almost no constipation, 3 . Unable to judge, 4 . Slightly constipated , 5. Constipation 5 . The difference between each subject before and after the intake of the experimental food was calculated and evaluated. For example, if the subject who was "5. Constipation" before the ingestion of the test food became "3. Unable to judge" after ingestion, the analysis was "the improvement of the whole intestine to "2 or more"." -37- (34)200826951 泽泽% m 01 m -4 1T Society. Spotted limbs 1 There are lower jaws J Right 21 Oral sweet start point End date Start date End date Start date End date Start曰 Start and end date 5 minutes (person) 0 0 1 0 0 0 Also 夂 口 口 itu 1 2 1 1 0 2 4 points (person) 2 10 4 17 1 8 1 3 6 10 5 8 3 points (person 9 14 8 7 1 1 1 11 2 7 5 7 6 3 2 points (person) 11 2 9 3 21 8 17 13 11 9 13 14 1 point (person) 6 2 8 1 5 1 1 7 3 5 1 4 1 Total (person) 28 28 28 28 28 28 28 28 28 28 Average score 2..3 3-1 1 2.5 3.4 1.9 | 2.9 2.0 2.6 2.5 3.0 Zo 2.4 28 2.9 Collagen egg _ • Neural guanamine · "With skin wrinkles (Forehead) Junze (mouth corner) Excitation (eye corner) Dry 起始 Start date End date Start date Start date End date Start 曰 End date Start 曰 End date Start latch soft 丨 {- p 5 points (person) 0 0 0 2 1 1 in It (2) 2 2 0 0 3 3 4 points (person) 5 9 10 9 Q 6 12 〇 15 0 13 11 15 3 points (person) 8 14 4 5 J 3 3 5 0 2 6 5 2 points (person) 11 4 9 9 1 λ 12 8 1U 8 10 10 8 5 1 point (person) 4 1 5 3 4 1 5 1 18 3 0 0 Total (person) 28 28 28 28 28 28 28 28 28 28 28 Average score 2.5 3.1 2.7 2.9 2.7 3.3 2.6 3.2 1.4 2.9 3.3 Z5 3.6 Through-bow reddish makeup Persistence edema constipation Resisting 昍颢 Start 曰 End date Start 曰 End date start曰 termination date start 曰 termination date start date end ^^0 termination date 5 minutes (person) 0 1 2 2 0 2 0 3 0 7 1 1 4 points (person) 2 5 11 10 5 16 11 15 0 5 3 6 3 points (person) 8 15 4 3 6 8 1 3 1 5 2 6 2 points (person) 14 6 7 11 15 2 15 5 17 8 12 13 1 point (person) 4 1 4 2 L 0 1 2 10 3 10 2 Total (person) 28 28 28 28 28 28 28 28 28 28 28 28 Average score 2.3 3.0 3.0 3.0 2.5 3.6 2.8 3.4 1.7 3.2 2.0 2.7 After ingesting the test foods of Examples 5 and 6, evaluate the knot of skin elasticity - 38- (35) 200826951 The results are shown in Figures 10 to 12. Fig. 1 is a subject whose "intestinal improvement is "1 or more"", and Fig. 11 is a subject whose "intestinal improvement is "2 or more"", and Fig. 1 is "the improvement of the whole intestine to "3 or more". The subject evaluated the skin elasticity. It is confirmed that the improvement effect on the whole intestine (stool) is also high, and the skin improvement effect is also high. The test food (containing collagen peptide and ceramide) which favored Example 6 had an excellent effect as the test food of Example 5. By ingesting the test foods of Examples 5 and 6, the skin elasticity and the improvement were improved (m Cheng and Ze. After ingesting the test foods of Examples 5 and 6, the results of evaluating the skin texture density were not as shown in Figs. 13 to 15. Fig. 13 For the subjects whose "intestinal improvement is "1 or more"", Figure 14 shows the subject of "intestinal improvement to "2 or more"". Figure 5 shows that "the whole intestine is improved to "3 or more"" The result of the evaluation of the skin texture density of the subject. It was confirmed that the improvement effect on the whole intestine (stool) was higher, and the skin improvement effect was also high. The test food of Example 4 (containing collagen peptide and neuropterin) was preferred. It is superior to the test food of Example 5. (Effects by ingesting the test foods of Examples 5 and 6 can increase skin texture density and improve skin texture. • According to the present invention, it is possible to provide high safety due to natural substances. Sexuality - and its skin-improving effect and/or therapeutic effect is high, and it contains a composition of lactic acid bacteria excellent in manufacturing characteristics and fermented milk. Further, it can provide a skin having an improved effect on the whole intestine and/or a therapeutic effect, skin Improve the effect and / Fermented milk with high therapeutic effect. [Simplified illustration] -39- (36) (36)200826951 [Fig. 1] Lactobacillus rhamnosus ATCC 5 3 1 0 3 to the group, Streptococcus thermophilus OLS 3 05 9 administered to the group, and non-initiated dermatitis group, the evolution map of dermatitis symptoms data during the test period (expressed as mean 値i: standard deviation . **Ρ<0·01,*Ρ<0· 05, multiple comparison of Dunnett). [Fig. 2] In the distilled water administration group, Lactobacillus rhamnosus ATCC 5 3 1 0 3 was administered to the group, Streptococcus thermophilus OLS 3 05 9 was administered to the group, and non-induced dermatitis group, 15 days later (day 15) Total IgE concentration in serum (expressed as mean 标准±standard deviation 。. * * P < 0.0 1 5 Multiple comparisons of Dunnett). [Fig. 3] In the steamed water retention group, Lactobacillus bulgaricus The OLL1073R-1 administration group and the Streptococcus thermophilus OLS 3 05 9 were administered to the group, and the evolution map of the dermatitis symptoms data during the test period (expressed by the mean soil standard deviation 。) **Ρ<0.01,*Ρ<〇· 〇5, Dunnett's multiple comparisons). Figure 4 is a graph of the total IgE concentration in the serum of the distilled water administration group, Lactobacillus bulgaricus OLL1073R-1 administered group, and Streptococcus thermophilus OLS3 05 9 group, 15 days later (day 15) (based on the average standard deviation of 値Said. **Ρ<〇·〇1, Dunnett's many comparisons). [Fig. 5] The administration group of fermented product prepared by Lactobacillus bulgaricus OLL1073R-1 and Streptococcus thermophilus OLS3059, and Lactobacillus delbrueckii subspecies bulgaricus MEP 1 70560 1 , and Streptococcus thermophilus -40- (37) (37) 200826951 MEP1705601 prepared fermented product group, unfermented product group and non-induced dermatitis and unfermented product group, the evolutionary map of dermatitis symptoms data during the test period (expressed by mean soil standard deviation 値**P<0.01, *P<0.05, multiple comparisons of Dunnett). [Fig. 6] Fermented milk prepared by Thermomyces elegans OLS3059 (Streptococcus thermophilus OLS3 05 9 ) was used as a test food, and the skin elasticity map of the subject. [Fig. 7] Fermented milk prepared by Streptococcus thermophilus OLS 3 05 9 (Streptococcus thermophilus OLS 3 05 9 ) was used as a test food, and the skin texture density map of the subject. [Fig. 8] A graph showing the observation results of the total number of rashes of the fermented milk prepared by Thermomyces elegans OLS3059 (Streptococcus thermophilus OLS3059) as a test food. [Fig. 9] Fermented milk prepared by Streptococcus thermophilus OLS 3059 (Streptococcus thermophilus OLS 3 05 9 ) was used as a test food, and the results of the evaluation of the effectiveness of the subjects and physicians were performed. [Fig. 10] The skin elasticity map of the fermented milk prepared by Streptococcus thermophilus OLS 3059 (Streptococcus thermophilus OLS 3 05 9 ) as a test food and the "intestinal improvement to "1 or more". [Fig. 11] A skin elasticity map of a fermented milk prepared by Streptococcus thermophilus OLS 3 059 (StrePtococcus thermophilus OLS3059) as a § test food product "the whole intestine is improved to "2 or more". [Fig. 12] Fermented milk prepared by Streptococcus thermophilus OLS 3 059 as a test food '-41 - (38) 200826951 "The improvement of the intestine is "3 or more"" Skin (Fig. 13) Fermented milk prepared by Streptococcus thermophilus OLS3 05 9 thermophilus OLS3059) The skin of the subject whose "intestinal improvement was "1 or more" was evaluated [Fig. 14] by Streptococcus thermophilus OLS3 05 9 Thermophilus OLS 3 05 9 ) The fermented milk prepared by the fermented milk is estimated to have "the whole intestine is improved to "2 or more"". The skin of the subject (Fig. 15) is fermented milk prepared by Streptococcus thermophilus OLS3 05 9 thermophilus OLS3059). The skin elasticity map of the subject whose intestine was improved to be more than 3". [Streptococcus: test food, texture density map. [Streptococcus: test food, texture density map. [Streptococcus: test food] texture density map. -42-
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