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詳目顯示區塊

第 30 筆
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論文名稱: | 乳酸菌細菌素抗菌作用及其在食品上應用之回顧
| Antimicrobial Activity of Lactic Acid Bacteria Bacteriocins and Its Application in Foods – A Review
研究生: | 龍湘美 Hsiang-Mei Lung
指導教授: | 潘崇良
學位類別: | 碩士
學校名稱: | 國立臺灣海洋大學
系所名稱: | 食品科學系
畢業學年度: | 92
語文別: | 中文
關鍵字: | 細菌素 bacteriocin
| 乳酸菌 lactic acid bacteria
全文點閱次數: | 22

全文影像連結:

論文全文(pdf)

論文頁數: | 222
摘要: | 摘 要
| 自然界中包括動、植物、昆蟲、及細菌等，均可以產生天然的抗菌蛋白質 (胜肽)，其主要作用為人體、動物疾病、植物疾病或是食品病菌之抗菌劑。由細菌所生產的抗菌蛋白質 (胜肽) 通稱為細菌素 (bacteriocin)。革蘭氏陽性菌及革蘭氏陰性菌均可生產各類細菌素，由乳酸菌所生產的細菌素依分子量大小可分為 class I (< 5 kDa)、class II (< 10 kDa)、及 class III (> 30 kDa)。其中對熱具安定性的乳酸菌細菌素可被區分為 (1) 胺基酸經轉譯後修飾者歸類在 class I 中，稱為 lantibiotic，包括 nisin 和 lacticin 等；(2) 胺基酸未經轉譯後修飾者被歸類在 class II 的 non-lantibiotics，包括 carnobacteriocin、enterocin、pediocin、及 sakacin 等。歸類在 class III 的乳酸菌細菌素，為對熱具敏感的大分子胜肽。無法以上述原則歸類者，屬於非典型 (atypical) 細菌素，如 bifidocin、leucocin B-TA33a、及 mesenterocin 52B 等。主要生產細菌素的乳酸菌有 Carnobacterium、Enterococcus、Lactobacillus、Lactococcus、Pediococcus、及 Streptococcus 等。乳酸菌細菌素主要抑制對象為革蘭氏陽性細菌，可抑制的菌種包括污染性的乳酸菌以及 Listeria、Bacillus、Clostridium 及 Sta. aureus 等食品病原菌營養細胞的生長及產孢菌孢子的萌發，故能廣泛應用在發酵或非發酵的乳品、肉品、魚製品、醃漬食品及穀類等食品系統上。屬於 class IIa 的 pediocin 家族則是由於其胺基酸排列的特性，對於 Listeria的菌株有特別強的抗菌作用。但這些乳酸菌細菌素單獨使用時對於革蘭氏陰性菌無抗菌的作用。添加化學防腐劑延長食品儲存期限已無法被現代的消費者接受，因此天然以及輕加工的食品現已普遍被重視，故自然產生的乳酸菌細菌素可以被大量生產和應用在食品中。乳酸菌細菌素的應用方式如做為發酵食品中生產細菌素的菌酛、直接添加純化的細菌素在食品製程中、或是附加在包覆材料上抑制食品表面的細菌等來做為食品防腐及品質改良的重要角色。雖然許多乳酸菌細菌素是從食品的乳酸菌中所分離和產生，但並非在所有的食品系統中均能展現有效的抗菌作用，因為在實際的應用上會因食品的物理和化學特性、環境條件、使用濃度、或作用時間等因素而影響該細菌素的活性；而且當這些細菌素所使用的濃度不足或作用時間不夠長時，會使目標菌產生抗性，例如 L. monocytogenes Scott A (ATCC 700302) 已經被確認會對 nisin 產生抗性 (nisin-resistance, Nisr)。為確保及加強這些乳酸菌細菌素的抗菌效果，以乳酸菌細菌素為基礎的多重防腐系統 (multiple–hurdle preservation system)，或是應用欄柵技術 (hurdle technology) 合併使用其他物理或化學防腐方法，提高對於食品病原菌的抗菌效果是必要的。這些防腐技術包括了添加螯合劑、使用溶菌素、提高鹽濃度、加入乳化劑、以及物理性的防腐 (如高靜水壓、溫度控制、調氣包裝、調整食品環境的 pH 值、加熱及低溫儲存等)，此外細菌素之間的合併使用，也可以對革蘭氏陰性菌如 E. coli、Salmonella、以及 Pseudomonas 等產生抗菌的能力。
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| Abstract
| Natural antimicrobial proteins (peptides) are produced by animals, plants, insects, and bacteria in nature. These natural peptides could be used as antimicrobial agents in human’s, animal’s, or plant’s diseases or on food pathogens. Antimicrobial proteins (peptide) produced by bacterium are called bacteriocin. Bacterioicn can be produced by Gram positive and Gram negitive bacteria. The bacteriocins produced by lactic acid bacteria are divided into class I (< 5 kDa), class II (< 10 kDa), and class III (> 30 kDa) according to their molecular weight. Heat-stable LAB bacteriocins whose amino acids are through post-translational modification are classified into class I, called lantibiotic, including nisin and lacticin. As to the LAB bacteriocins that their amino acids are not through post-translational modification are classified into class II, called non-lantibiotics, including carnobacteriocin, enterocin, pediocin, and sakacin. Class III are heat-labile peptides of large molecule weight. Atypical bacteriocins, which can not be classified by those princrples, such as bifidocin, leucocins B-TA33a, and mesenterocin 52B. LAB bacteriocins are majorly produced by Carnobacterium, Enterococcus, Lactobacillus, Lactococcus, Pediococcus, and Streptococcus. The most inhibition spectra of LAB bacteriocins are restricted Gram-positive bacteria, in which are contaminated LAB, Listeria, Bacillus, Clostridium, and Sta. aureus. Mostly of the inhibition exhibited on the growth of vegetative cells and the germination of endospores. Therefore, LAB bacteriocins could be utilized on the food systems of dairy product, meat, fishery product, pickled product, and cereal product, whether it was fermented or not. The pediocin familiy, which were classified as class IIa, showed more stronger antimicrobial effect on Listeria than other bacterial genera, that may be due to their specific amino acid sequences. Mostly, LAB bacteriocins did not present antimicrobial effect on Gram negetive bacteria while used alone. In order to prolong food shelf life, the chemical preservative were used traditionally by the producer, but foods preserved by this way such will not be accepted by modern consumers. The naturally and minimally processed foods are become more popular than before. The also naturally produced LAB bacteriocins were bulkly produced by LAB and being applied in varieus foods. The important way that LAB bacteriocins applied including: (A) the starter can produced bacteriocins by itself; (B) the purified bacteriocins were added during food manufacture; (C) or the bacteriocins are coating on the wrapping material and being released after food packaging to acehieve food preservation and quality improvement. Although many LAB bacteriocins derived from different sources of food, but the antimicrobial effect could not be monitored among the different food systems by these LAB foods. This could be dued to the antimicrobial effect of LAB bacteriocins as used in real food systems are influenced by its the physical and chemical properties, environmental conditions, concerntrations used, and duration of treatment. As the concerntration of LAB bacteriocins or the time of treating is not enough high or long, the target bacteria could develop the resistance to the bactericin used. For example, L. monocytogenes Scott A (ATCC 700302) has been identified to be resistant to nisin (Nisr). To assure and enhance the antimicrobial effect of LAB bacteriocins, LAB bacteriocins should be employed as the foundation of the multiple–hurdle technology preservation system or to be used in the hurdle as combined with other physical and chemical agents. These measures should improve the antimicrobial effect of LAB bacteriocins on the common foodborne pathogens. Such preserving technology includes adding chelating agent, emplying lysozyme, increasing salt concentration, using emulsifier, and other physical technology as hydrostatic pressure, controlled temperature, modified atmosphere packaging, adjusted pH value of the food environment, heating, and cooling treatment. In addition, the combination different LAB bacteriocins also could inhibit the Gram-negative bacteria such as E. coli, Salmonalla, and Pseudomonas.
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論文目次: | 目 錄
| 目錄 ………………………………………………………………… i
| 表目錄 ……………………………………………………………… v
| 圖目錄 ……………………………………………………………… vi
| 附錄 ………………………………………………………………… vii
| 名詞縮寫 …………………………………………………………… viii
| 中文摘要 …………………………………………………………… x
| 英文摘要 …………………………………………………………… xii
| Ⅰ、前言 …………………………………………………………… 1
| Ⅱ、乳酸菌分類 …………………………………………………… 3
| (Ⅰ) 雙歧桿菌屬 (Bifidobacterium) ………………………… 4
| (Ⅱ) 肉品桿菌屬 (Carnobacterium) ………………………… 4
| (Ⅲ) 腸球菌屬 (Enterococcus) ………………………………… 5
| (Ⅳ) 乳酸桿菌屬 (Lactobacillus) …………………………… 5
| (Ⅴ) 乳酸球菌屬 (Lactococcus) ……………………………… 7
| (Ⅵ) 明串球菌屬 (Leuconostocs) …………………………… 7
| (Ⅶ) 酒球菌屬 (Oenococcus) ………………………………… 8
| (Ⅷ) 小球菌屬 (Pediococcus) ………………………………… 8
| (Ⅸ) 有孢子乳桿菌屬 (Sporolactobacillus) ………………… 9
| (Ⅹ) 鏈球菌屬 (Streptococcus) ……………………………… 9
| (ⅩⅠ) 四體球菌屬 (Tetragenococcus) ………………………… 9
| (ⅩⅡ) 徘徊球菌屬 (Vagococcus) ……………………………… 10
| (ⅩⅢ) Weissella ………………………………………………… 10
| (ⅩⅣ) 其他 ……………………………………………………… 11
| Ⅲ、乳酸菌一般特性及抗菌作用 ………………………………… 13
| (Ⅰ) 一般特性 ………………………………………………… 13
| (Ⅱ) 抗菌物質種類……………………………………………… 15
| 1. pH 值及有機酸 ……………………………………… 15
| 2. 過氧化氫 (H2O2) ……………………………………… 16
| 3. 二氧化碳 (CO2) ……………………………………… 17
| 4. 細菌素 ………………………………………………… 17
| 5. 洛德因系統 (reuterin system) ………………………… 18
| 6. 雙乙醯 (diacetyl) ……………………………………… 18
| Ⅳ、細菌素 (抗菌胜肽) 種類與特性 …………………………… 20
| (Ⅰ) 一般特性 ………………………………………………… 21
| (Ⅱ) 分類 ……………………………………………………… 21
| 1. Class I …………………………………………………… 22
| 2. Class II ………………………………………………… 22
| 3. Class III ………………………………………………… 23
| Ⅴ、乳酸菌細菌素特性、分類與作用機制 ………………………… 24
| (Ⅰ) 一般特性與分類 ………………………………………… 24
| (Ⅱ) 乳酸菌細菌素分類、作用機制與種類 …………………… 26
| 1. Class I (lantibiotic) ……………………………………… 27
| (1) 一般特性 …………………………………………… 27
| (2) 分類與抗菌機制 …………………………………… 28
| A. 分類 ……………………………………………… 28
| B. 抗菌機制 ………………………………………… 29
| (3) 種類 ………………………………………………… 30
| A. Nisin ……………………………………………… 30
| B. Lacticin 3147 ……………………………………… 31
| C. Lacticin 481 ……………………………………… 32
| D. Lactocin S ………………………………………… 32
| E. Mutacin …………………………………………… 32
| F. 其他 class I 乳酸菌細菌素 ……………………… 33
| 2. Class II (non-lantibiotic) ……………………………… 33
| (1) 一般特性 …………………………………………… 34
| A. Class IIa …………………………………………… 36
| B. Class IIb …………………………………………… 36
| C. Class IIc …………………………………………… 36
| (2) 抗菌機制 …………………………………………… 37
| (3) 種類 ………………………………………………… 37
| A. Class IIa …………………………………………… 37
| (A) Pediocin ……………………………………… 37
| (B) Carnobacteriocin ……………………………… 39
| (C) Divercin ……………………………………… 39
| (D) Enterocin ……………………………………… 40
| (E) Sakacin ………………………………………… 41
| (F) 其他 class IIa 乳酸菌細菌素 ……………… 42
| B. Class IIb …………………………………………… 43
| 3. Class III 乳酸菌細菌素 ……………………………… 44
| 4. 非典型 (atypical) 乳酸菌細菌素 …………………… 44
| (Ⅲ) 乳酸菌細菌素抗菌目標 …………………………………… 45
| 1. 對革蘭氏陰性菌與革蘭氏陽性菌細胞之影響 ……… 45
| 2. 對產孢細菌細胞之影響 ……………………………… 45
| (Ⅳ) 乳酸菌細菌素抗菌試驗與抗菌範圍 …………………… 46
| 1. 乳酸菌細菌素和其他因子抗菌效果 ………………… 46
| 2. 乳酸菌細菌素抗菌範圍 ……………………………… 52
| (Ⅴ) 抗乳酸菌細菌素菌株產生之機制及影響 ……………… 52
| 1. Nisr 抗菌機制 ………………………………………… 53
| 2. 結合其他抗菌因子對Nisr細菌之影響 ……………… 55
| Ⅵ、乳酸菌細菌素在食品上之應用 ……………………………… 59
| (Ⅰ) 生產乳酸菌細菌素菌株之分離來源 ……………………… 59
| 1. 發酵食品來源 ………………………………………… 60
| (1) 乾酪製品 …………………………………………… 60
| (2) 魚、肉類製品 ……………………………………… 62
| (3) 蔬果製品 …………………………………………… 63
| (4) 穀類及其他製品 …………………………………… 63
| 2. 非發酵食品來源 ……………………………………… 64
| (1) 乳製品、魚、肉製品 ……………………………… 64
| (2) 蔬果類製品 ………………………………………… 64
| (3) 飲料與酒 …………………………………………… 65
| (4) 穀類及其他非食品來源 …………………………… 65
| (Ⅱ) 乳酸菌細菌素在食品上之應用 …………………………… 66
| 1. 發酵食品 ……………………………………………… 68
| (1) 乾酪製品 …………………………………………… 68
| (2) 肉製品與海鮮 ……………………………………… 72
| (3) 蔬菜製品 …………………………………………… 74
| (4) 飲料與酒類 ………………………………………… 75
| (5) 穀類製品 …………………………………………… 76
| 2. 非發酵食品 …………………………………………… 76
| (1) 乳製品 ……………………………………………… 76
| (2) 肉製品 ……………………………………………… 77
| (3) 魚及海鮮製品 ……………………………………… 83
| (4) 蔬菜、水果及沙拉 ………………………………… 85
| (5) 飲料與酒類 ………………………………………… 85
| (6) 豆及穀類製品 ……………………………………… 86
| (7) 包裝材料 …………………………………………… 87
| (8) 其他用途 …………………………………………… 89
| Ⅵ、乳酸菌細菌素應用於食品產業上的未來展望 ……………… 90
| Ⅶ、參考文獻 ……………………………………………………… 91
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| 表 目 錄
| 表一、革蘭氏陽性菌細菌素分類 …………………………………… 139
| 表二、乳酸菌細菌素生產菌株及胺基酸相關特性 ………………… 141
| 表三、乳酸菌細菌素濃度和其他因子之抗菌效果 ………………… 147
| 表四、乳酸菌細菌素之抗菌範圍 …………………………………… 152
| 表五、不同來源之乳酸菌細菌素生產菌株 ………………………… 158
| 表六、乳酸菌細菌素做為食品防腐之應用 ………………………… 162
| 表七、使用乳酸菌細菌素做為食品欄柵技術中部分增加抗菌活性 的因子 ……………………………………………………… 166
| 表八、乳酸菌細菌素在不同食品系統之防腐應用 ………………… 172
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| 圖 目 錄
| 圖一、數種乳酸菌 lantibiotics 初級結構 ………………………… 181
| 圖二、數種非乳酸菌 lantibiotics 初級結構 ……………………… 182
| 圖三、Nisin 誘使形成類似楔子孔洞之模型 ……………………… 183
| 圖四、Class I 及 class IIa 細菌素胺基酸 N 端序列 …………… 184
| 圖五、圖解 Class IIa 細菌素的結構模式與目標細胞膜預期反應的位置 ……………………………………………………… 185
| 圖六、圖解革蘭氏陽性菌與革蘭氏陰性菌細胞壁之比較 ………… 186
| 圖七、某些食品防腐劑對孢子膨脹、出芽或成長的影響 ………… 187
| 圖八、電子顯微鏡下 enterocin CRL35 對 L. monocytogenes 細胞的影響 ………………………………………………… 188
| 圖九、掃描式電子顯微鏡下 enterocin 81 對 L. monocytogenes 細胞的影響 ………………………………………………… 189
| 圖十、溶菌素及 nisin 對 Sta. aureus 細胞膜的影響 …………… 190
| 圖十一、溶菌素及 nisin 對於 Lb. sake 的影響 ………………… 191
| 圖十二、Pediocin AcH 與靜水壓對於 Leu. mesenteroides 細胞的 溶菌影響 ………………………………………………… 192
| 圖十三、Nisin 對於 L. monocytogenes 抗性菌株及野生菌株之作用模式 …………………………………………………… 193
| 圖十四、Enterocin P 對於 Ent. faecium 敏感或具抗性細胞的作用 模式 ……………………………………………………… 194
| 圖十五、Enterocin 在乾酪上對 L. innocua LMG 的抑菌效果 … 195
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| 附 錄
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| 附錄Ⅰ、乳酸鏈球菌素在各國的使用狀況 ………………………… 196
| 附錄Ⅱ、動物、植物及真菌天然抗菌胜肽 ………………………… 198
| 附錄Ⅲ、洋菜孔洞擴散法 ………………………………………… 200
| 附錄Ⅳ、不同濃度 nisin 對於革蘭氏陽性菌形成的抗菌環效果 201
| 附錄Ⅴ、不同濃度 nisin 添加 EDTA 對於革蘭氏陽性菌形成的抗菌環 …………………………………………………… 202
| 附錄Ⅵ、不同濃度 nisin 對於革蘭氏陰性菌形成的抑菌環效果 203
| 附錄Ⅶ、不同濃度 nisin 添加 EDTA 對於革蘭氏陰性菌形成的抑菌環 …………………………………………………… 204
| 附錄Ⅷ、野生株細胞及對 nisin 具抗性細胞在 nisin 存在或缺少時對細胞上磷脂質組成的影響 ………………………… 205
| 附錄Ⅸ、乳製品分類 ……………………………………………… 206
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