TWI465565B - 含有雙叉乳酸桿菌屬細菌之醱酵食品及其製造方法 - Google Patents
含有雙叉乳酸桿菌屬細菌之醱酵食品及其製造方法 Download PDFInfo
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- TWI465565B TWI465565B TW095117130A TW95117130A TWI465565B TW I465565 B TWI465565 B TW I465565B TW 095117130 A TW095117130 A TW 095117130A TW 95117130 A TW95117130 A TW 95117130A TW I465565 B TWI465565 B TW I465565B
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- bifidobacterium
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Description
本發明係關於含有雙叉乳酸桿菌屬細菌之醱酵食品，更詳細而言，含有高濃度之活的雙叉乳酸桿菌屬細菌，並且保存後之存活性優異之醱酵食品及其製造方法。
雙叉乳酸桿菌屬細菌係與代表乳酸桿菌屬細菌等之乳酸菌，同樣地顯示具有改善腸內菌叢、改善排便性、改善腸道功能、防禦感染、免疫賦活、預防癌症等之各種功效。因此認為此等微生物係藉由改善腸內環境，幫助人體健康。
雙叉乳酸桿菌屬細菌為發揮如此效果，於醱酵乳等之製品中，要求保持高生菌數。然而，因為雙叉乳酸桿菌屬細菌，通常是嫌氣性細菌，所以缺乏存活性，尤其於氧存在下，急速地死亡。
因此，使用雙叉乳酸桿菌屬細菌之醱酵乳等製品中，為提供其存活性為目的，揭示例如使用蔗糖或山梨糖醇(右旋葡萄糖醇，D-glucitol)之方法(專利文獻1)，使用赤蘚醇之方法(專利文獻2)，使用乳糖醇(Lactitol)之方法(專利文獻3)等。
另外，雖亦提及為提供具有活力狀態之雙叉乳酸桿菌屬細菌予消費者，由充填剛製造後之含有雙叉乳酸桿菌屬細菌之醱酵物，於不透氧性包材所構成之容器，完全阻斷
與氧接觸等之手段，但使用如此之不透氧性之容器，於廢棄處理面上或成本面上留有許多問題，該利用之自由度受到限制。
因此，至今有幾個報告進行檢討於好氣的條件下，維持雙叉乳酸桿菌屬細菌之存活性之手段。具體上，為提高雙叉乳酸桿菌屬細菌之存活性，揭示使用N-乙醯葡糖胺、泛酸或雙泛醯硫乙胺、泛醯硫氫乙胺、胜類、乳果糖之方法。然而，上述方法所使用之素材本身價格較高，利用於食品時對風味的影響大等，亦包含存活性之效果，要求更優異的素材。
另外，雙叉乳酸桿菌屬細菌之存活性係容易受到菌株、製品(飲食品等)之pH值、作為甘味料配合之糖質及溶存氧量等之影響，以改善此為目的，進行檢討關於添加酵母或乳酸菌、併用維生素C、製品容器之材質等。然而，為因應近年多樣化之消費者嗜好性或更加提昇存活性，要求可作為更優異存活性用途使用之素材。
【專利文獻1】特公昭57-4291號公報
【專利文獻2】特許第2577692號
【專利文獻3】特許第3261571號
因此，本發明之課題係提供可期待各種生理效果，可
提高活的雙叉乳酸桿菌屬細菌之保存存活性，得到即使使用於飲食品等時，亦不影響風味之優異的新穎素材，並且，由該素材而改善存活性，含有高濃度之活的雙叉乳酸桿菌屬細菌之醱酵食品。另外，本發明亦以提供於製造含有雙叉乳酸桿菌屬細菌之醱酵食品時，改善其保存時之存活性之方法為該課題。
本發明者為解決上述課題，努力研究的結果，發現由配合特定之植物萃取物，可得到安定並維持雙叉乳酸桿菌屬細菌於保存後之生菌數之醱酵食品，而完成本發明。
亦即，本發明係以含有1種或2種以上選自薑黃、魚腥草、杜仲、米糠、柿葉、紫蘇、丁香、肉桂及甜茶所成群之植物素材之萃取物，與雙叉乳酸桿菌屬細菌為特徵之醱酵食品。
另外，本發明係以含有1種或2種以上選自由酸萃取薑黃、魚腥草、杜仲、米糠、柿葉、紫蘇、丁香、肉桂及甜茶所成群之植物素材之萃取物，尤其以pH4.0以下之條件所萃取之上述萃取物與雙叉乳酸桿菌屬細菌為特徵之醱酵食品。
進一步，本發明係於製造含有雙叉乳酸桿菌屬細菌之醱酵食品，以任何階段配合1種或2種以上選自薑黃、魚腥草、杜仲、米糠、柿葉、紫蘇、丁香、肉桂及甜茶所成群之植物素材之萃取物為特徵之含有雙叉乳酸桿菌屬細菌
之醱酵食品之製造方法。
更進一步，本發明係以配合1種或2種以上選自薑黃、魚腥草、杜仲、米糠、柿葉、紫蘇、丁香、肉桂及甜茶所成群之植物素材之萃取物為特徵之雙叉乳酸桿菌屬細菌之存活性改善方法。
本發明之醱酵食品所含有之1種或2種以上選自薑黃、魚腥草、杜仲、米糠、柿葉、紫蘇、丁香、肉桂及甜茶所成群之植物素材之萃取物，對於雙叉乳酸桿菌屬細菌，具有優異的存活性改善效果，而且幾乎不影響風味。因此，使用上述萃取物之本發明之醱酵食品係風味良好，即使長期保存，仍無損生菌數，為優良的健康增進者。
本發明之醱酵食品係以含有1種或2種以上選自由酸萃取薑黃、魚腥草、杜仲、米糠、柿葉、紫蘇、丁香、肉桂及甜茶所成群之植物素材之萃取物(以下亦簡稱為「萃取物」)為特徵者。
成為上述萃取物原料之植物素材中，薑黃(Turmeric)係薑科薑黃屬植物之鬱金(Curcuma longa L.)或薑黃(Curcuma aromatica Salisb.)之根莖。本發明之此等薑黃屬植物中，以薑黃為宜。已知薑黃具有提昇肝功能、預防宿醉、抑制胃酸分泌、改善腸胃障礙等之作用。
魚腥草(Houttuynia cordata Thunb.)係魚腥草科魚腥草屬植物。雖可使用地上部份之全草或枝，用以自魚腥草取得萃取物，但以使用地上部份之全草為宜。已知魚腥草具有抑制黏膜發炎之作用。
杜仲(Eucommia ulmoides Oliv.)係杜仲科杜仲屬植物。雖可使用葉或枝，用以自杜仲得到萃取物，但以使用葉為宜。已知杜仲具有調整血壓、緩和壓力、預防生活習慣病之作用。
所謂米糠(Rice bran)係指自禾本科稻屬植物之水稻(Oryza sativa)所得之除去「稻穀」外皮(稻殼)之種子(糙米)之果皮、種皮澱粉層、胚芽。已知米糠具有增強免疫力、預防脂肪肝等之作用。
柿葉係柿樹科柿樹屬植物之柿(Diospyros Kaki Thumb)、豆柿(Diospyros lotus L.)或豆柿(Diospyros lotus L.var.glabra Makino)的葉子。本發明中此等柿樹屬植物中，以柿為宜。柿葉具有抑制打噴嚏、鼻塞、流鼻水等之作用。
紫蘇(Perilla)係紫蘇科紫蘇屬植物之紫蘇(Perilla frutescens(L.)Britton var.acuta kudo)、綠紫蘇(Perilla frutescens(L.)Britton var.acuta kudo forma viridis Makino)或紫蘇(Perilla frutescens(L.)Britton var.crispa(Thunb)Decne)。本發明中此等紫蘇屬植物中係以紫蘇尤佳。另外，雖可使用葉、枝、種子，用以自此等紫蘇得到萃取物，但以使用葉子為宜。已知此紫蘇
具有抗過敏、降低血糖值、使肌膚年輕等之作用。
丁香(Clove)係桃金娘科蒲桃屬植物之丁香(丁子香)(Syzygium aromaticum(L.)Merr.EtPerry或Eugenia caryophyllata Thunb)之花蕾。已知此丁香具有防腐、子宮收縮、減少牙疼等之作用。
肉桂(Cinnamon)係樟科肉桂屬之錫蘭肉桂(Cinnamomum zeylanicum Nees)或中國肉桂(Cinnamomum cassia Blume)之樹皮。此等肉桂屬植物中，以錫蘭肉桂為宜。已知肉桂具有抗菌、溫熱身體、解熱、鎮痛、活化消化器官、改善感冒各種症狀、改善消化不良、停止腹瀉、停止嘔吐等之作用。
甜茶(Rubus suavissimus S.Lee(Rosaceae))係薔薇科懸鈎子屬植物。雖可使用葉或莖，用以得到來自甜茶之萃取物，但以使用葉子尤佳。近年來，甜茶具有抗炎症或抗過敏作用而備受矚目。
為自上述植物素材得到萃取物，可將1種以上之上述植物素材，直接或因應需要而施以洗淨、脫皮、乾燥、破碎等之處理後，以溶媒萃取即可。此等萃取物係可使用分別萃取各植物素材所得之萃取物中之1種，亦可混合2種以上使用。另外，亦可使用混合多種植物素材後，萃取此等所得之混合萃取物。此等萃取物中，以柿葉萃取物及甜茶萃取物為宜。
作為製造上述萃取物所使用之溶媒，可舉例如水或乙醇等之碳數1至5之低級醇、乙酸乙酯、甘油、丙二醇等
之有機溶媒。此等溶媒係可混合2種以上作為混合溶媒使用。此等溶媒中，以水或水-低級醇等之水性溶媒尤佳。
另外，使用上述溶媒之萃取方法，雖無特別的限制，但因為可有效地自各植物素材萃取提高對雙叉乳酸桿菌屬細菌之存活性改善效果之成份，即使此萃取物之添加量係微量，仍可得到充份的效果，所以以酸萃取法為宜。另外，此酸萃取係於pH4.0以下，以於pH3.0至4.0之酸性條件下進行尤佳。進行此酸萃取時，為調整溶媒之pH所使用之酸成份，只要為酸性物即可，並無特別的限制，皆可使用，但作為適合者，可舉例如檸檬酸、蘋果酸、酒石酸、琥珀酸、乳酸、醋酸等之有機酸。
進一步，關於使用上述溶媒之萃取物之萃取條件，雖無特別的限制，例如60℃以上，120℃以下之溫度，以80℃以上，100℃以下之溫度為宜，進行萃取處理30至60分鐘左右尤佳。
如此所得之萃取物係可以原本的狀態，使用所萃取之萃取物，並且，亦可使用將所得之萃取物，由超濾、離心分離等手段而精製、濃縮之濃縮萃取物，或將其再由噴霧乾燥或冷凍乾燥等之手段而乾燥之粉末狀萃取物。
本發明之醱酵食品中上述萃取物之使用量係依雙叉乳酸桿菌屬細菌之種類等而所得效果會發生差異，以實驗確定後設定為宜，例如以水萃取所得之萃取物(以下稱為「水萃取物」)時，作為糖度約為Brix10之萃取物，為0.01質量%至10.0質量%(以下僅稱為「%」)左右，以
0.1%至50%左右為宜。另外，以酸萃取所得之萃取物(以下稱為「酸萃取物」)時，作為糖度約為Brix10之萃取物，為0.001%至10%左右，以0.01%至1.0%左右為宜。
本發明之醱酵食品中，即使水萃取物或酸萃取物之使用量多於10%時，並不能得到與雙叉乳酸桿菌屬細菌之添加量成比例以上之存活性改善效果，並且，因為對醱酵食品風味造成影響，所以不適宜。另外，相反地水萃取物之使用量少於0.01%時或酸萃取物之使用量少於0.001%時，並不能充份地得到對雙叉乳酸桿菌屬細菌之存活性改良效果，所以不適宜。
另一方面，製造本發明之醱酵食品所使用，包含於其中之雙叉乳酸桿菌屬細菌，只要係屬於雙叉乳酸桿菌屬細菌之微生物即可，該種類雖無特別的限制，但該適合者，可舉例如已知作為人類腸內菌叢之主要者之短雙叉乳酸桿菌(Bifidobacterium breve)、長雙叉乳酸桿菌(Bifidobacterium longum)、嬰兒雙叉乳酸桿菌(Bifidobacterium infantis)、(Bifidobacterium adolescentis)、青春雙叉乳酸桿菌(Bifidobacterium bifidum)、鏈雙叉乳酸桿菌(Bifidobacterium catenulatum)、假鏈雙叉乳酸桿菌(Bifidobacterium pseudocatenulatum)、Bifidobacterium angulatum、具有來自分離自人類腸內之Bifidobacterium gallicum、利用於食品之乳雙叉乳酸桿菌(Bifidobacterium lactis)、動物雙叉乳酸桿菌(
Bifidobacterium animalis)等。此等雙叉乳酸桿菌屬細菌中，就與上述萃取物併用時之存活性改善效果方面上，以短雙叉乳酸桿菌、青春雙叉乳酸桿菌(Bifidobacterium bifidum)、長雙叉乳酸桿菌尤佳。
進一步，本發明之醱酵食品中，由萃取物對於雙叉乳酸桿菌屬細菌之存活性改善效果，即使含有雙叉乳酸桿菌屬細菌以外之多種微生物時，仍可得之。因此，本發明之醱酵食品中，除了雙叉乳酸桿菌屬細菌以外，亦可含有凱氏乳酸桿菌(Lactobacillus casei)、嗜酸乳酸桿菌(Lactobacillus acidophilus)、瑞士乳酸桿菌(Lactobacillus helveticus)、唾液乳酸桿菌(Lactobacillus salivarius)、加氏乳酸桿菌(Lactobacillus gasseri)、乳酸桿菌屬酵母菌(Lactobacillus fermentum)、洛德乳酸桿菌(Lactobacillus reuteri)、捲縮乳酸桿菌(Lactobacillus crispatus)、德氏乳酸桿菌保加利亞亞種(Lactobacillus delbrueckii subsp.bulgaricus)、Lactobacillus delbrueckii subsp.delbueckii、約氏乳酸桿菌(Lactobacillus johnsonii)等之乳酸桿菌屬細菌、嗜熱鏈球菌(Streptococcus thermophilus)等之鏈球菌屬細菌、乳酸乳球菌(Lactococcus lactis subsp.lactis)、乳酸乳球菌(Lactococcus lactis subsp.cremoris)、胚芽乳球菌(Lactococcus plantarum)、棉子糖乳球菌(Lactococcus raffinolactis)等之乳球菌屬細菌、糞腸球菌(Enterococcus faecalis)、尿腸球菌(Enterococcus
faecium)等之腸內球菌屬細菌等之乳酸菌。
另外，尤其本發明之醱酵食品中之萃取物對於雙叉乳酸桿菌屬細菌之存活性改善之效果，該醱酵食品中之雙叉乳酸桿菌屬細菌之菌數為1×107/ml以上，尤其1×108/ml以上，可得到明顯的效果。
本發明之醱酵食品係於使用上述雙叉乳酸桿菌屬細菌(以及因應需要之乳酸菌)之醱酵食品製造中，於任何階段添加上述萃取物以外，可依據傳統已知之使用雙叉乳酸桿菌屬細菌之醱酵食品之製造方法而製造。例如，於殺菌處理脫脂奶粉之前或之後，添加萃取物，接種所需之雙叉乳酸桿菌屬細菌等而培養，將其均質化處理而得醱酵乳基料，接著添加、混合另外調製之糖漿溶液，再添加香料等，完成最終製品即可。另外，亦可將殺菌處理之脫脂奶粉溶液，接種所需之雙叉乳酸桿菌屬細菌等而培養，將其均質化處理而得醱酵乳基料，接著添加、混合另外調製之糖漿溶液及萃取物，再添加香料等而完成最終製品。
另外，本發明中所謂之醱酵食品，係指乳類等由省令所決定之醱酵乳、乳製品、乳酸菌飲料等之飲料或硬式優酪乳、軟式優酪乳、原味優酪乳，進一步包含醱酵乳(kefir)、乳酪等。另外，於本發明之醱酵食品中，亦包含利用各種乳酸菌之飲食品，例如原味型態、加味型態、水果型態、甜味型態、軟式型態、飲料型態、固體(硬式)型態、冷凍型態等之醱酵乳、乳酸菌飲料、醱酵乳(kefir)、乳酪等。
進一步，本發明之醱酵食品，因應需要，除了糖漿等甘味料以外，可配合其他各種食品素材，例如各種糖質、增黏劑、乳化劑、各種維生素等之任何成份。作為此等食品素材，具體上可配合蔗糖、葡萄糖、果糖、巴拉金糖(palatinose)、海藻糖、乳糖、木糖、麥芽糖等之糖質、山梨糖醇、木糖醇、赤蘚糖、乳糖醇、還原巴拉金糖、還原水飴、還原麥芽糖水飴等之糖醇、阿斯巴甜、索馬甜(thaumatin)、蔗糖素、醋磺內酯鉀、甜菊等之高甜度甘味料、瓊脂、明膠、鹿角菜膠、瓜爾豆膠、三仙膠、果膠、刺槐豆膠、結蘭膠、羧甲基纖維素、大豆多糖類、褐藻酸丙二醇酯等之各種增黏(安定)劑、蔗糖脂肪酸酯、甘油脂肪酸酯、聚甘油脂肪酸酯、山梨糖醇酐脂肪酸酯、卵磷脂等之乳化劑、奶酪(cream)、奶油、酸乳酪等之乳脂肪、檸檬酸、乳酸、醋酸、蘋果酸、酒石酸、葡糖酸等之酸味料、維生素A、維生素B類、維生素C、維生素E類等之維生素類、鈣、鎂、鋅、鐵、錳等之礦物質成分、優酪乳系、莓系、柳橙系、花梨系、紫蘇系、橘櫞系、蘋果系、薄荷系、葡萄系、杏桃系、梨、布丁饀、桃子、哈密瓜、香蕉、熱帶水果、香草系、紅茶、咖啡系等之香料類。
如上述說明之本發明之醱酵食品中，由添加混合萃取物，與至今之含有雙叉乳酸桿菌屬細菌之醱酵食品相比較，改善保存時之存活性之理由至今雖未明確，但推測係因為於各植物素材所萃取之萃取物，含有許多礦物質，此對
於雙叉乳酸桿菌屬細菌之存活性改善效果，有很大的幫助。
以下係舉實施例，更詳細地說明本發明，但本發明並不局限於此等實施例等。
對於薑黃(Curcuma longa L.之根莖)、魚腥草(Houttuynia cordata Thunb.)之地上部全草、杜仲(Eucommia ulmoides Oliv.)之葉、米糠(自水稻(Oryza sativa)所得之除去「稻穀」外皮(稻殼)之種子(糙米)之果皮、種皮澱粉層、胚芽)、柿葉(Diospyros Kaki Thumb之葉)、紫蘇(Perilla frutescens(L.)Britton var.acuta Kudo之葉)、丁香(Syzygium aromaticum(L.)Merr.Et Perry之花蕾)及肉桂(Cinnamomum zeylanicum Nees之樹皮)，分別施以脫皮、破碎等之處理後，以90℃之熱水(各原料之10倍量)，萃取60分鐘，調製薑黃萃取物、魚腥草萃取物、杜仲萃取物、米糠萃取物、柿葉萃取物、紫蘇萃取物、丁香萃取物及肉桂萃取物。將此等分別以蒸發器濃縮，調整各萃取物成糖度10。
調製以12%之脫脂奶粉溶液作為基本培養基，於其中分別添加各1%之調製成糖度10之薑黃萃取物、魚腥草萃取物、杜仲萃取物、米糠萃取物、柿葉萃取物、紫蘇萃取物、丁香萃取物或肉桂萃取物之滅菌培養基。於此等滅菌培養基中，接種1%之短雙叉乳酸桿菌之引酵物，以37℃培養至pH為4.8左右。
將培養結束時以及以10℃保存14天後培養物中之生菌數，以TOS(transgactosyl-oligosacchandes)培養基(Yokult藥品工業(股)社製)測定。以此測定之生菌數為基礎，依據下式，求取雙叉乳酸桿菌屬細菌之殘存率。進而，以基本培養基之殘存率作為基準，添加各萃取物之培養基之存活性改善率係依下式求出。其結果係如表1所示。
如表1所示，於添加薑黃、魚腥草、杜仲、米糠、柿葉、紫蘇、丁香及肉桂之各萃取物之培養基，與基本培養基相比較，認為改善對雙叉乳酸桿菌屬細菌存活性之效果。
於實施例1之萃取物製造中，改使用以水及檸檬酸，調整成pH3.0、4.0、5.0之水溶液以取代熱水以外，以相同條件處理柿葉，製造各糖度10之柿葉萃取物。於添加1%之所得各萃取物之12%之脫脂奶粉培養基，接種1%之短雙叉乳酸桿菌(YIT10001)之引酵物，以37℃進行培養至pH成4.8左右。培養結束時以及所得之培養物以10℃保存14天後之短雙叉乳酸桿菌之生菌數，係與實施例2同樣地測定。另外，與實施例2同樣地操作，自生菌數求出殘存率，進而求出存活性改善率。其結果係如表2所示。
如表2所示，認為對雙叉乳酸桿菌屬細菌之存活性改善效果，使用調製成pH 5.0以下之萃取物，尤其調製成pH4.0以下之萃取物，有明顯的改善趨勢。
對於薑黃(Curcuma longa L.之根莖)、魚腥草(Houttuynia cordata Thunb.)之地上部全草、杜仲(Eucommia ulmoides Oliv.)之葉、米糠(自水稻(Oryza sativa)所得之除去「稻穀」外皮(稻殼)之種子(糙米)之果皮、種皮澱粉層、胚芽)、柿葉(Diospyros Kaki Thumb之葉)、紫蘇(Perilla frutescens(L.)Britton var.acuta Kudo之葉)、丁香(Syzygium aromaticum(L.)Merr.Et Perry之花蕾)及肉桂(Cinnamomum zeylanicum Nees之樹皮)，分別施以脫皮、破碎等之處理後，使用以水及檸檬酸，調整成pH4.0之溶液(10倍量)，以與實施例1相同的條件萃取，調製薑黃萃取物、魚腥草萃取物、杜仲萃取物、米糠萃取物、柿葉萃取物、紫蘇萃取物、丁香萃取物及肉桂萃取物。將此等分別以蒸
發器濃縮，調整成糖度10。
以15%之脫脂奶粉溶液作為基本培養基，於其中分別添加1%之實施例4之調製成糖度10之薑黃萃取物、魚腥草萃取物、杜仲萃取物、米糠萃取物、柿葉萃取物、紫蘇萃取物、丁香萃取物或肉桂萃取物，調製培養基。於各培養基中，接種1%之雙叉乳酸桿菌屬細菌之引酵物，以37℃培養48小時。培養結束時以及以10℃保存14天後之所得培養物之短雙叉乳酸桿菌之生菌數，係與實施例2同樣地測定。另外，與實施例2同樣地操作，自生菌數求出殘存率，進而求出存活性改善率。其結果係如表3所示。
另外，培養中係使用短雙叉乳酸桿菌、青春雙叉乳酸桿菌及長雙叉乳酸桿菌作為雙叉乳酸桿菌屬細菌。
如表3所顯示，此等萃取物對於各雙叉乳酸桿菌屬細菌之存活性所造成的影響，雖依菌株的種類而多少有些差
異，但認為對全部的菌株有影響。認為尤其薑黃萃取物、魚腥草萃取物、杜仲萃取物、柿葉萃取物有顯著的效果。
準備以15%之脫脂奶粉培養基作為基本培養基，於其中分別添加0.1%之實施例4所製造之各種萃取物之試驗培養基。將此等培養基加熱殺菌後，於各培養基，接種1%之短雙叉乳酸桿菌(YIT10001)之引酵物、各0.1%之乳酸乳球菌(Lactococcus lactis)(YIT2027)及嗜熱鏈球菌(Streptococcus thermophilus)(YIT2021)之引酵物，以35℃，進行24小時培養，而得培養物。將此培養液以15MPa均質化物之40重量份，加入60重量份之以100℃殺菌5分鐘之10%砂糖溶液，再添加0.1%之優酪乳香料((股)Yakult Material社製)，製造乳製品。對於此乳製品，由5名經驗豐富之品評員，進行味覺試驗時，認為任一種乳製品與含有使用基本培養基所得培養物之對照品皆無差異。
另外，培養基所添加之各種萃取物，對於基本培養基並不造成風味上之影響，亦有非常調和之評價，並確認即使用於乳製品等之飲食品所使用之培養，亦不使風味劣化。
使用以水及檸檬酸調整成pH4.0之水溶液(10倍量)，以與實施例1相同的條件，分別調製柿葉萃取物。將此等以蒸發器濃縮，調整成糖度10。
於15%之脫脂奶粉培養基，添加0.001至10%範圍之(1)所調製之糖度10之柿葉萃取物，以100℃殺菌60分鐘，調製乳酸菌培養培養基。此培養培養基中，接種1%之短雙叉乳酸桿菌(YIT10001)之引酵物、各0.1%之乳酸乳球菌(YIT2027)及嗜熱鏈球菌(YIT2021)之引酵物，以35℃，進行24小時培養。將此培養液以15MPa均質化物之40重量份，加入60重量份之以100℃殺菌5分鐘之10%之砂糖溶液，再添加0.1%之優酪乳香料((股)Yakult Material社製)，製造乳製品。對於所得之乳製品，由5名經驗豐富之品評員，依據下述基準進行風味評估。另外，製品化後以及以10℃保存所得製品14天後之雙叉乳酸桿菌屬細菌之生菌數，係與實施例2同樣地測定。另外，與實施例2同樣地操作，自生菌數求出殘存率，進而求出存活性改善率。其結果係如表4所示。
如表4所示，由添加0.01%左右之柿葉萃取物，認為具有對於雙叉乳酸桿菌屬細菌之存活性改善效果。另一方面，即使添加10%左右之柿葉萃取物於培養基，並不能得到更優異的效果，反而有來自柿葉之風味影響製品風味之趨勢。另外，由柿葉萃取物對於雙叉乳酸桿菌屬細菌之存活性改善效果，由酸萃取比由水萃取者以較少的添加量而
效果明顯。
使用以水及檸檬酸調整成pH4.0之水溶液(10倍量)，以與實施例1相同的條件，分別調製柿葉萃取物。將此等以蒸發器濃縮，調整成糖度10。
將15%之脫脂奶粉培養基，以100℃，殺菌60分鐘後，接種1%之短雙叉乳酸桿菌(YIT10001)之引酵物、各0.1%之乳酸乳球菌(YIT2027)及嗜熱鏈球菌(YIT2021)之引酵物，以35℃，進行24小時培養，得到培養物。之後，與實施例2同樣地測定培養物之乳酸菌數。接著，將此培養液以15MPa均質化物之40重量份，加入60重量份之以100℃殺菌5分鐘之10%之砂糖溶液者，混合0.1%之所調製之各柿葉萃取物之水溶液於10%之砂糖溶液，以100℃殺菌5分鐘者，添加0.1%之優酪乳香料((股)Yakult Material社製)，製造乳製品。對於所得之乳製品，以與實施例7相同的基準，進行風味評估。另外，製品化後以及以10℃保存所得製品14天後之雙叉乳酸桿菌屬細菌之生菌數，係與實施例2同樣地測定。進一步，與實
施例2同樣地操作，自生菌數求出殘存率，進而求出存活性改善率。其結果係如表5所示。
由表5認為未添加柿葉萃取物於培養基，即使於得到培養物後添加，仍可得到對雙叉乳酸桿菌屬細菌之存活性改善效果。另外，於得到培養物後，即使添加柿葉萃取物，亦不造成乳製品風味之不良影響。
對甜茶(Rubus suavissimus S.Lee(Rosaceae))葉子，施以脫皮、破碎等之處理後，以90℃的熱水(甜茶葉之10倍量)，萃取60分鐘，調製甜茶萃取物。將所得之培養物以蒸發器濃縮，調整萃取物糖度成10。
以12%之脫脂奶粉溶液作為基本培養基，於其中添加1%之實施例9所製造之調整糖度為10之甜茶萃取物，調製滅菌培養基。於此滅菌培養基，接種1%之短雙叉乳酸
桿菌之引酵物，以37℃培養至pH成4.8左右。
將培養結束時以及以10℃保存14天後培養物中之生菌數，以TOS(transgactosyl-oligosacchandes)培養基(Yogurt藥品工業(股)社製)測定。以此測定之生菌數為基礎，依據下式，求取雙叉乳酸桿菌屬細菌之殘存率。進而，以基本培養基之殘存率作為基準，添加甜茶萃取物之培養基之存活性改善率係依下式求出。其結果係如表6所示。
如表6所示，添加甜茶萃取物之培養基，與基本培養基相比較，認為對雙叉乳酸桿菌屬細菌，具有優異的存活性改善效果。
於實施例9之製造萃取物中，除了使用水及調整pH成3.0、4.0、5.0之檸檬酸水溶液以取代熱水以外，以同樣的條件處理甜茶，製造各糖度10之甜茶萃取物。於添加1%所得之各萃取物之12%脫脂奶粉培養基，接種1%之短雙叉乳酸桿菌(YIT10001)之引酵物，以37℃進行培養至pH成4.8左右。培養結束時以及以10℃保存14天後之所得培養物之短雙叉乳酸桿菌生菌數，係與實施例10同樣地測定。另外，與實施例10同樣地操作，自生菌數求出殘存率，進而求出存活性改善率。其結果如表7所示。
如表7所示，認為對雙叉乳酸桿菌之存活性改善效果係甜茶萃取物萃取溶媒之pH愈低，則有愈高之趨勢。
對甜茶(Rubus suavissimus S.Lee(Rosaceae))葉
子，施以脫皮、破碎等之處理後，使用以水及調整成pH4.0之檸檬酸水溶液(10倍量)，以與實施例9相同條件萃取，調製甜茶萃取物。將所得之培養物以蒸發器濃縮，調整成糖度10。
以15%之脫脂奶粉作為基本培養基，於其中添加1%之實施例12之調整糖度為10之甜茶萃取物，調製培養基。於此培養基，接種1%之各種雙叉乳酸桿菌屬細菌之引酵物，以37℃培養至pH成4左右。培養結束時以及所得之培養物以10℃保存14天後之雙叉乳酸桿菌屬細菌之生菌數，係與實施例10同樣地測定。另外，與實施例10同樣地操作，自生菌數求出殘存率，進而求出存活性改善率。其結果如表8所示。
另外，培養係使用作為雙叉乳酸桿菌屬細菌之短雙叉乳酸桿菌、青春雙叉乳酸桿菌及長雙叉乳酸桿菌。
如表8所示，由甜茶萃取物對雙叉乳酸桿菌屬細菌之存活性改善效果雖依菌株而多少有些差異，但認為幾乎全部的菌株具有效果。另外，依據菌株的種類，亦確認可得到縮短培養時間的效果。
準備以20%之脫脂奶粉培養基作為基本培養基，於其中添加0.1%之實施例12所製造之甜茶萃取物之試驗培養基。將此培養基加熱殺菌後，接種1%之短雙叉乳酸桿菌(YIT10001)之引酵物、各0.1%之乳酸乳球菌(YIT2027)及嗜熱鏈球菌(YIT2021)之引酵物，以35℃，進行24小時培養，得到培養物。將此培養物以15MPa均質化物之40重量份，加入60重量份之以100℃殺菌5分鐘之10%之砂糖溶液，再添加0.1%之優酪乳香料((股)Yakult Material社製)，製造乳製品。對於此乳製品，由5名經驗豐富之品評員，進行味覺試驗時，認為與含有使用基本培養基所得之培養物之對照品並無差異。
另外，培養基所添加之甜茶萃取物，對於基本培養基不造成風味上之影響，亦有非常調和之評價，並確認即使用於乳製品等之飲食品所使用之培養，亦不使風味劣化。
使用水及調整成pH4.0之檸檬酸水溶液(10倍量)，以與實施例9相同的條件，調製甜茶萃取物。將此以蒸發器濃縮，調整成糖度10。
於20%之脫脂奶粉培養基，添加0.001至10%範圍之(1)所調製之糖度10之甜茶萃取物，以100℃殺菌60分鐘，調製培養基。此培養基中，接種1%之短雙叉乳酸桿菌(YIT10001)之引酵物、各0.1%之乳酸乳球菌(YIT2027)及嗜熱鏈球菌(YIT2021)之引酵物，以35℃，進行24小時培養，得到培養物。之後，與實施例10同樣地測定培養物之乳菌數。接著，於此培養物以15MPa均質化物之40重量份，加入60重量份之以100℃殺菌5分鐘之10%之砂糖溶液，再添加0.1%之優酪乳香料((股)Yakult Material社製)，製造乳製品。對於所得之乳製品，由5名經驗豐富之品評員，依據下述基準進行風味評估。另外，製品化後以及以10℃保存14天後所得製品之雙叉乳酸桿菌屬細菌之生菌數，係與實施例10同樣地測定。進一步，與實施例10同樣地操作，自生菌數求出殘存率，進而求出存活性改善率。其結果係如表9所示。
如表9所示，添加0.01%左右之甜茶萃取物，具有對雙叉乳酸桿菌屬細菌之存活性改善效果。另一方面，認為即使添加10%左右之甜茶萃取物，並不能得到更優異的效果，反而有影響製品風味之趨勢。另外，對於雙叉乳酸桿菌屬細菌之存活性改善效果，由酸萃取比由熱水萃取之甜茶萃取物顯示更明顯的效果。
使用水及調整成pH4.0之檸檬酸水溶液(10倍量)，以與實施例9相同的條件，調製甜茶萃取物。將此萃取物以蒸發器濃縮，調整成糖度10。
將20%之脫脂奶粉培養基以100℃殺菌60分鐘後，接種1%之短雙叉乳酸桿菌(YIT10001)之引酵物、各0.1%之乳酸乳球菌(YIT2027)及嗜熱鏈球菌(YIT2021)之引酵物，以35℃，進行24小時培養，得到培養物。接著，將此培養物以15MPa均質化，於該40重量份中，加入60重量份之以100℃殺菌5分鐘之10%之砂糖溶液，或混合(1)所調製之0.1%甜茶萃取物之水溶液於10%之砂糖溶液，以100℃殺菌5分鐘者，再添加0.1%之優酪乳香料((股)Yakult Material社製)，製造乳製品。對於所得之乳製品，與實施例2同樣地測定製造終了時以及以10℃保存14天後所得製品之雙叉乳酸桿菌屬細菌之生菌數。進一步，與實施例10同樣地操作，自生菌數求出殘存率，進而求出存活性改善率。其結果如表10所示。
由表10之結果顯示，甜茶萃取物對雙叉乳酸桿菌屬細菌之存活性改善效果，與該萃取物之添加時間無關，皆可獲得。
本發明之醱酵食品係即使含有雙叉乳酸桿菌屬細菌，長期保存風味仍然良好，而且因為係維持生菌數者，所以可利用於增進健康。
Claims (4)
- 一種醱酵食品，其特徵為含有將甜茶於pH4.0以下之條件下進行酸萃取而得者與雙叉乳酸桿菌屬細菌。
- 如申請專利範圍第1項之醱酵食品，其中更含有選自乳酸桿菌屬(Lactobacillus)、鏈球菌屬(Streptococcus)、乳球菌屬(Lactococcus)、腸內球菌屬(Enterococcus)之乳酸菌者。
- 一種含有雙叉乳酸桿菌屬細菌之醱酵食品之製造方法，其特徵為於含有雙叉乳酸桿菌屬細菌之醱酵食品之製造中，於任何之階段配合將甜茶於pH4.0以下之條件下進行酸萃取而得者。
- 一種雙叉乳酸桿菌屬細菌之存活性改善方法，其特徵為配合將甜茶於pH4.0以下之條件下進行酸萃取而得者。
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