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研究生:郝敏雯
研究生(外文):Miin-Win Hao
論文名稱:益生菌在保健食品之回顧與展望
論文名稱(外文):Review and Future Development ofProbiotics in Health Foods
指導教授:潘崇良
指導教授(外文):Chorng-Liang Pan
學位類別:碩士
校院名稱:國立海洋大學
系所名稱:食品科學系碩士在職專班
學門:農業科學學門
學類:食品科學類
論文種類:學術論文
論文出版年:2002
畢業學年度:90
語文別:中文
論文頁數:196
中文關鍵詞:乳酸菌益生菌食品安全性特性健康效益加工食品
外文關鍵詞:Lacticc acid bacteriaprobioticFood SafetyHealth effectsProcessing food
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摘 要
發酵乳中產酸的乳酸桿菌被認為可抑制宿主大腸中的害菌產生毒性物質,因而推論其可保護腸道並促進宿主健康且得以延年益壽,此觀念提出時明顯的僅專注於乳品中的乳酸菌,後續的研究逐漸轉向存在於宿主腸道中具有存活能力的菌株,最近的定義則為利用具生理調節機能活性之死菌和活菌及其代謝物質等的混和產物,食品中可被允許使用的益生菌,主要屬於Bifidobacterium、Lactobacillus、Lactococcus、Streptococcus thermophilus、Enterococcus、Saccharomyces 等菌群。乳製品中有益健康的微生物--益生菌 (probiotics) 於1965年最早被描述為多種的原生動物 (protozoa) 於對數生長期間,可產生導致其他原生動物生長期延長之物質,到2001年益生菌的定義擴展為含足以改變宿主胃腸道菌相的活菌量及有益宿主健康的活菌組合食品,及至今漸被接受的益生菌是無論活菌、死菌、菌體萃取物,甚或是微生物代謝產物等均為益生菌的應用形態,食品使用的菌株應源自人體並歸類為食品級。對於益生菌的鑑定技術發展如利用選擇性培養基檢測共生菌群中的不同益生菌株,使用鑑定培養基tryptone proteose peptone yeast extract with Prussian blue (TPPYPB agar),可在同一培養盤上生長的益生菌因菌落形態的差異,而可有效檢測四種益生菌株的各別生菌數。利用分子生物技術分類時,可藉多元分類 (polyphasic taxonomy) 方法結合形態、生化、生理特性與分子生物技術等來鑑別,利用16S/23S rRNA基因序列資料庫繪製不同益生菌株的演進分析樹圖 (phylogenetic trees or dendrograms)。由於宣稱具生理效益之添加益生菌食品導入市場,而發展出針對此類食品更專精的安全性評估之研究領域,此類食品也常需要結合病理學家等多種專業人員共同驗證。近年來有些感染的案例出現如菌血症和心內膜炎等皆證明與乳酸桿菌有關,故使用新菌株做為食品或醫藥的成份之前,應審慎評估和測試該益生菌株的安全性及添加益生菌食品的安全性效益。篩選益生菌株時應考量其應用之重要因素,如(1)易製成活菌濃縮品,並可工業規模量產,(2)於製造及貯存過程中,與食品載體共存時仍具存活能力,(3)於宿主胃腸道中存活及促進宿主健康的能力等。為確保益生菌株促進宿主健康的能力,評估益生菌株保健功能的測試應包括菌株的表現型 (phenotypic) 和基因型 (genotypic) 安定性的項目如:(a)質體安定性;(b)醣類及蛋白質的利用型態 (c)在酸及膽鹽環境下的耐受力、存活及生長能力;(d)腸道上皮細胞之吸附特性;(e)產生抗微生物之物質及抑制病原菌生長能力等。一般認為應攝食足夠活菌量的益生菌食品,始能達到調整腸道菌相促進宿主健康,益生菌在人類腸道中的吸附特性、代謝活性、生長形態、生長菌量及宿主的各別差異等均會影響其對宿主的作用,當益生菌於宿主的腸道中定殖後,可能利用其競爭優勢,於宿主胃腸道中與病原菌競爭營養源;並可藉由封鎖吸附受器和/或產生短鏈脂肪酸 (short-chain fatty acids, SCFAs),及產生細菌素等抑菌物質抑制病原菌生長等作用能力,而達成預防感染、調節免疫等增進宿主健康。為使益生菌能存活於食品加工處理後的食品中,應審慎選擇益生菌種,並考量菌株的特性及菌株間的交互作用,控制加工及貯藏的條件如種菌量、培養溫度和培養時間等,維持益生菌株的安定性及活性。益生菌目前已廣泛使用於發酵食品、嬰兒配方食品、水果飲料、甜奶飲料等,其中以優酪乳及乳酪為使用較為普遍的食品形態,益生菌之應用產品型態有膠囊、錠劑及不同的發酵乳製品,或是添加入食品中構成該類食品的健康特性,如在休閒食品之糖果點心、餅乾、冰淇淋等之中添加具益生菌功能的微生物。在食品應用上,益生菌的研究方向將朝向選擇新的及對宿主健康更具專一性的菌株例如針對年齡層、健康者、疾病者等之個體差異性;益生菌的分類和鑑定工作上,應就現有技術改善其可靠性及使其趨向於易操作、和低成本;期望產業和學術界能合作進行對於食品、胃腸道、人類健康等領域之整合研究;臨床研究上應積極研究益生菌於人類胃腸道中的作用機制,發展診斷工具和生物標誌以增進對於宿主消化管道中各種生化及免疫反應的瞭解,未來應發展含益生菌的非乳製食品如黃豆類、穀類及休閒類等之食品。
關鍵字:乳酸菌、益生菌 (probiotics)、食品安全性、特性、健康效益、加工食品。
Abstract
It has been known that the lactic-producing lactobacilli in fermented dairy products can inhibit potentially pathogenic bacteria toxin formation in the large intestine of host and is inferred that having the effect of prevention and protection which can improve health and prolong life. Although the ideas were originally related to lactic acid bacteria in dairy products, the interest of other scientists soon turned to lactic acid bacteria of intestinal origin i.e., the living microorganisms. Recent work is further to define that as the products made from the cell preparations, components of viable cultures and non-viable cultures, or metabolites that have beneficial effect on the health of the host. Probiotic preparations nowadays used in food mainly belong to Bifidobacterium, Lactobacillus, Lactococcus, Streptococcus thermophilus, Enterococcus, and Saccharomyces species. Probiotic, the microorganism that has beneficial effects in fermented dairy products, was coined in 1965 and used to describe several species of protozoa that can produce substances to prolong the logarithmic phase in other species during their logarithmic phases of growth. In 2001 the definition is widened to as the foods or products containing enough amounts of live microorganisms that can positively influence the microflora of the colon and improve the health of the host when consumed in adequate amounts. Gradually the definition does not restrict the use of probiotics. Not only the whole microbial cells but also parts of cells including viable cultures, non-viable cultures, cells extracts, or metabolites of these microorganisms can be accepted as the applied forms of probiotics. Besides, the strains of probiotic used in food should be attributed to food grade organism and human origin. Several technologies are used for distinguishing probiotics from bacteria. The enumeration method selects a set of culture media to perform the enumeration of probiotic when it appears together with bacteria of lactic acid starters in fermented dairy products. The approaches are simple and time-saving and can offer a good cell recovery for the microorganisms. The taxonomy and physiology of probiotics can be implemented by using the polyphasic taxonomy that combines morphological, biochemical and physiological characteristics with molecular-based phenotypic and genomic techniques. Comparing the sequence of rRNA is currently the most powerful and accurate technique for determining the degree of phylogenetic relation of microorganisms. Based on the available information on the sequences of 16S/23S rRNA, the phylogenetic trees can be created. Due to the claimed beneficially physiological effects are beyond those of good nutrition without adverse effects, conventional toxicology and safety evaluation is of limited value in the safety evaluation of probiotic microorganisms. Therefore, a multidisciplinary approach involving the contribution of pathologists and microbiologists is necessary. Furthermore, a few rare cases of infection have so far been associated with lactic acid bacteria. Safety implications should be taken into consideration before using new strain to make composition food or medicine. A potential probiotic bacteria should fulfill the following criteria: (1) easy to prepare as viable culture concentrate on a large industrial scale, (2) able to survive during preparation and storage of the carrier foods, and (3) able to survive in the intestinal ecosystem of the host. To ensure the ability of improving health effects, the initial screening and selection of probiotics should include the stability test of phenotype and genotype which takes account of (a) plasmid stability; (b) carbohydrate and protein utilization patterns; (c) the ability to tolerate, survive, and grow in acid and the bile metabolism; (d) the adhesion properties of intestinal epithelium; (e) the ability to produce antimicrobial substances and antibiotic resistance patterns. The adhesive properties, metabolism, and morphology of probiotics grown in intestinal contents or intestinal mucus may affect health effects. The adherence to mucosal surfaces contributes to the efficacy of a probiotic. Probiotics indigenous strains might contribute to the prevention of infection by pathogenic micro-organisms by blocking attachment sites and/or producing short-chain fatty acids (SCFAs) that inhibit the growth of pathogenic organisms. The properties of fermented products depend on the strain, inoculation level, temperature, and time. The manufacturing process thus should consider carefully the effects on the selection and characteristics of starter cultures and the interactions between different strains to maintain the vitality and stability. Nowadays probiotics are widely used in fermented foods including infant formula, fruit drinks, and sweet milk. Fermented milk and cheese are the most common foods with probiotics. Probiotic cultures may be used in special shapes like capsules or tablets, or they may be used in the production of a large variety of fermented food products. In some cases the cultures may be added to a food to contribute specific probiotic or functional properties. With innovation in a variety of foods and drinks such as snack, biscuits, ice creams, and confectionery the incorporation of probiotic strains should not impact negatively on product quality. Future research on probiotic bacteria will center on selecting new and more specific strains for the well being of the host age group, healthy populations, and disease specific. The work of simplicity and liability to categorize and identify probiotic should be improved under the cost. The future research trends will be to inter-link the expertise and scientific knowledge on food, gastrointestinal tract (GI-tract) functionality, and human health. In the clinical research, develop diagnostic tools and biomarkers for their assessment will help studying the mechanisms of action of probiotics in the GI-tract. To develop technology for non-dairy probiotic applications (i.e., bean, cereal based materials and snack) will be essential for the probiotic industry.
Keywords: Lactic acid bacteria; Probiotic; Food safety; Health effects; Processing food
目 錄
目錄………………………………………………….…..………………….i
名詞縮寫………………………………………………………….….….…iv
表目錄………………………………………………..…..………………….v
圖目錄……………………………………………………………………....vi
中文摘要……………………………………………………….……….…..vii
英文摘要………………………………….…………………………….…...ix
壹、前言…………………………………………..……………………….…1
貳、益生菌之定義…………………………………..…………………..…...3
參、益生菌的鑑定…………………………………….…………….…….…5
肆、益生菌的安全性……………………………...…………………..……11
一、抗生素抗性…………………………………………………….….12
二、代謝活性…………………………………………………………..13
三、毒性…………………………………………….………….………15
四、感染性………………………………………………………….….16
五、使用於食品上的安全性…………………………………………..17
伍、益生菌之特性………………………………………………….….……19
一、胃腸道中之存活能力…………………………………………...…19
(一)、耐胃酸、膽鹽………………………………….……………20
1、耐胃酸…………………….………………...……….…20
2、耐膽鹽…………………………………..……..…….…21
(二)、吸附……………………………………..…………………..24
1、益生菌之吸附機制……………….…………...…….....25
2、益生菌的吸附能力………………………………..…...27
(1)、吸附於似人類腸道Caco-2細胞………………....28
(2)、吸附於似人類腸道粘膜…………………………..30
(3)、安全性……………………………………………..32
(三)、存活能力…………………………….…………..……….….32
二、產生抗微生物的物質……………………………………………..34
(一)、抑菌能力………………………………………..………..….35
陸、益生菌的健康效益…………………….…………..…………………..38
一、乳糖不耐症狀…………………………………………...…………38
二、免疫調整……………………………………………………..….…40
(一)、刺激免疫過程………………………………….……..…….42
(二)、非特異性免疫………………………………….…....….…..43
(三)、細胞激素…………………………………………….…..….44
(四)、體液免疫…………………………………………….…..….45
(五)、細胞免疫………………………………………...…..…..….47
(六)、免疫缺乏症患者…………………………..………...…..….48
(七)、非活性菌株的免疫刺激……………………..……….…….48
(八)、生理活性胜月太………………………………...……….…….50
三、防癌的角色……………………………….……………....……….51
(一)、可能的防癌機制……………………………..….…...….….51
(二)、增強宿主免疫能力……………………….…….…...…..….52
(三)、降低毒性………………………………….….……..….…...53
(四)、降低腸道中有害酵素的活性……………..….……....…….54
四、胃腸道疾病之預防及治療………..…….……….………………..57
(一)、改善腸道菌相…………………………..….…………...…..58
(二)、增強免疫預防感染…………..…....……….…..…….….….60
(三)、胃腸道炎、腹瀉及功能不良…………….…….……..…....62
(四)、胃潰瘍…………...………………………….….……...……64
(五)、抑制腸道中的病原菌生長……………….…….……..……65
五、預防心血管疾病…………………….…………...………….….…67
(一)、降低血清中膽固醇………..………………………………..68
(二)、降低血壓………………………………………………...….70
(三)、抗氧化………………………………………………………71
六、其他潛在的作用…………………………..…….……….…..….…74
(一)、泌尿道、陰道炎………………………………...………….75
(二)、過敏…………………………………….……….….……….76
(三)、重症患者………………………………..…………….…….77
(四)、其他…………………………………..………..……...…….77
柒、食品加工系統中之益生菌……………………..………………………80
一、發酵菌種………………………………………...…………………80
(一)、菌株交互作用…….………………………………...…..…...81
(二)、加工特性……………………………………….………....…82
二、胞外多醣……………………………………..…………………….85
三、減少衛生上的危害………………………………………...………87
(一)、抑菌活性………………………………..……………….….88
(二)、抗氧化能力……………………………………..……..……90
四、加工及貯藏中的安定性………….……...…………….…….……92
(一)、乳製品………………………………………..…….……….92
(二)、非乳製品…………………………………………..………..95
五、增加食品的消化利用及營養價值………………………………..96
(一)、增進蛋白質消化利用……………………………..……..…97
(二)、促進維生素、礦物質吸收代謝………………..….……….98
捌、益生菌食品……………………………………………….…………..100
一、發酵乳製品………………………………………………..…...…101
(一)、優酪乳…………………………………….…………..……102
(二)、乾酪 (cheese)……………………………………….…..…105
二、發酵肉製品…..….………………………………………..……....106
(一)、發酵香腸………………………………………….….……107
(二)、魚肉發酵香腸………………………………….….………108
三、發酵蔬果…...….……………………….………………….….…..109
四、烘焙食品.…………….…………………………….………..……111
五、機能食品.………………………………....…………….…….…..111
六、休閒食品….……………………………………..……….……….113
七、醫療製劑…....………………………...……………………..……114
玖、未來的展望……………………………………………………...…….117
拾、參考文獻…….………………………………………………...………118
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