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詳目顯示區塊

第 22 筆
| (以作者名查詢海洋大學圖書館館藏系統)
| (以作者名查詢全國圖書書目資訊網)
| (以作者名查詢全國博碩士系統)
論文名稱: | 乳酸鏈球菌素與 EDTA 對腸炎弧菌等革蘭氏陰性菌抗菌效果之探討
| Studies on the antibacterial effect of nisin and EDTA against several Gram-negative bacteria as Vibrio parahaemolyticus
研究生: | 黃文鈴 Wen-Lin Huang
指導教授: | 潘崇良 Chong-Liang Pan
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
學號: | T91420030
畢業學年度: | 93
語文別: | 中文
關鍵字: | 乳酸鏈球菌素 nisin
| 腸炎弧菌 Vibrio parahaemolyticus
| 抗菌效果 antibacterial effect
全文點閱次數: | 14

全文影像連結:

論文全文(pdf)

論文頁數: | 88
摘要: | 摘　要
|
| 乳酸鏈球菌素 (nisin) 為由 Lc. lactis subsp. lactis 所分泌的一種抗菌多胜肽 (polypeptide)。Nisin 已被證明對於革蘭氏陽性菌及病原菌具較廣的抑制效果。當 nisin 與食品中常用的抗氧化劑和螯合劑 EDTA 合併使用時，即可有效地減少 E. coli、Salmonella 等革蘭氏陰性菌之含量。本研究為探討 E. coli BCRC 11549、Sal. choleraesuis subsp. choleraesuis BCRC 10744、V. parahaemolyticus isolates 1、5、10、或 16，在起始 pH 值為 4.0、5.0、或 6.0 之適當培養液中，配合 26oC 或 37oC 的培養溫度，合併使用 50 □g mL-1 nisin 及 20 mM EDTA 所呈現之被抑制效果。在以洋菜孔洞擴散測試法所得實驗結果顯示 nisin 對於指標菌 Str. thermophilus BCRC 12268 最小抑制濃度為 100 ppm。單獨使用不同濃度的 nisin 或 EDTA 對於 V. parahaemolyticus isolates 1、5、10、及 16 皆無抑菌環產生。E. coli BCRC 11549、Sal. choleraesuis subsp. choleraesuis BCRC 10744、V. parahaemolyticus isolates 1、5、10、或 16，在起始 pH 值為 4.0、5.0、或 6.0 之適當培養液中，於 26oC 或 37oC 培養 12 hr 後，未加抗菌物質的控制組所含菌數皆高於實驗組。
| 至於 V. parahaemolyticus isolates 1、5、10、或 16 在起始 pH 值為 4.0、5.0、或 6.0 下以 37oC 培養，並以 50 □g mL-1 nisin 與 20 mM EDTA 處理 1-6 hr後的殘存率為 0%。而V. parahaemolyticus isolates 1、10、或 16在起始 pH 值為 4.0 和 26oC 下以相同濃度 nisin 與 EDTA 處理 3-6 hr 後所觀察到的殘存率亦為 0%。其中以 V. parahaemolyticus isolate 1 最為敏感於起始 pH 值為 4.0 培養液中於 37oC 下培養 1.0 hr 後菌數即降到未能檢出。E. coli BCRC 11549、及 Sal. choleraesuis subsp. choleraesuis BCRC 10744 在 37oC 起始 pH 值為 4.0或 6.0培養時，50 □g mL-1 nisin 與 20 mM EDTA 作用後殘存生菌數為 3.1 及 3.3 log CFU/mL。在前述實驗中，E. coli BCRC 11549、Sal. choleraesuis subsp. choleraesuis BCRC 10744 以及 V. parahaemolyticus isolates 1、5、10、或 16 之控制組與實驗組生菌數 (log CFU/mL)對數值之最大差異分別為 4.4、5.5 以及 9.0、8.8、8.8、或 8.88。
|
| Abstract
|
| Nisin is an antimicrobial polypeptide produced by certain strains of Lc. lactis subsp. lactis.  It has been demonstrated that nisin exhibits a wide range of inhibitory effects against Gram-positive bacteria and pathogens.  When nisin combination with EDTA, a chelating agent commonly used as an antioxidant in the food industry, has been shown to be effective for reducing populations of Gram-negative bacteria, such as E. coli and Salmonella.  In this study, 50 □g mL-1 nisin combined with 20 mM EDTA were used to testing the antibacterial effect against E. coli BCRC 11549, Sal. choleraesuis subsp. choleraesuis BCRC 10744, and V. parahaemolyticus isolates 1, 5, 10, or 16.  The testing conditions were initial pH of broth at 4.0, 5.0, or 6.0 as well as incubation temperature at 26oC or 37oC.  Experiment results from agar well diffusion method showed that the minimum inhibitory concentration (MIC) of nisin against indicator strain, Str. thermophilus BCRC 12268, was 100 ppm.  Inhibition zone was not observed when various concentrations of nisin or EDTA was used individually to testing against V. parahaemolyticus isolates 1, 5, 10, or 16.  While E. coli BCRC 11549, Sal. choleraesuis subsp. choleraesuis BCRC 10744, V. parahaemolyticus isolates 1, 5, 10, or 16 was treated with nisin (50 □g mL-1) and EDTA (20 mM) under initial broth pH at 4.0, 5.0, or 6.0 and incubated at 26oC or 37oC for 12 hr, viable cell counts of experimental groups were lower than the control group.
| The antibacterial effect of 50 □g mL-1 nisin and 20 mM EDTA against V. parahaemolyticus isolates 1, 5, 10, or 16 was 100% as incubated in initial broth pH at 4.0, 5.0, or 6.0 and incubated at 37oC for 1-6 hr.  The survival rates of V. parahaemolyticus isolates 1, 10, and 16 were 0% as treated by 50 □g mL-1 nisin and 20 mM EDTA under initial broth pH at 4.0 and incubated at 26oC for 3-6 hr..  Among them, V. parahaemolyticus isolate 1 was the most sensitive one as its survival rate showed 0% after 1.0 hr incubation.  While E. coli BCRC 11549 and Sal. choleraesuis subsp. choleraesuis BCRC 10744 were tested by 50 □g mL-1 nisin and 20 mM EDTA for their surviving rarte under initial broth pH at 4.0 or 6.0 and incubated at 37oC, the viable cell count showed as 3.1 and 3.3 log CFU/mL, respectively.  The largest differences of viable cell count (log CFU/mL) between control group and experiment group (with nisin and EDTA treatment) observed in above experiments of E. coli BCRC 11549, Sal. choleraesuis subsp. choleraesuis BCRC 10744, V. parahaemolyticus isolates 1, 5, 10, or 16 were 4.4, 5.5, and 9.0, 8.8, 8.8, or 8.88, respectively.
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論文目次: | 目 錄
| 目    錄    i
| 表 目 錄    iv
| 圖 目 錄    v
| 中文摘要    vii
| 英文摘要    ix
| 一、前言    1
| 二、文獻整理    3
| I.  乳酸菌之定義、特性、及抗菌作用    3
| I-1.  乳酸菌之定義及特性    3
| I-2.  乳酸菌之抗菌作用    4
| II.  乳酸鏈球菌素 (nisin)    7
| II-1.  細菌素之發現及定義    7
| II-2.  Nisin 之發現    8
| II-3.  Nisin之介紹    9
| II-3-1.  Nisin之結構    9
| II-3-2.  Nisin之特性    10
| II-3-3.  Nisin之抗菌特性及機制    11
| II-3-4.  Nisin生產之影響因子    13
| II-3-4-1.  內因性因子    13
| II-3-4-2.  外因性因子    13
| II-3-5.  Nisin 抗菌活性的檢測    16
| II-3-6.  Nisin 對革蘭氏陰性菌與革蘭氏陽性菌
| 細胞之影響    17
| II-4.  EDTA 對革蘭氏陰性菌抗菌作用之機制    17
| II-5.  食品中常見的革蘭氏陰性病原菌    19
| II-6.  Nisin在食品加工上之應用    20
| 三、材料與方法    26
| I.  實驗材料    26
| I-1.  實驗菌株    26
| I-2.  培養基    26
| I-3.  實驗藥品    26
| I-4.  儀器設備    27
| II.  實驗方法    27
| II-1.  菌株之保存、活化、及生長曲線    27
| II-1-1.  菌株的保存    27
| II-1-2.  菌株之活化與接種菌源之製備    27
| II-1-3.  菌株之培養與計數    28
| II-2.  Nisin 及 EDTA 試劑之配製    28
| II-2-1.  乳酸鏈球菌素 (nisin) 溶液之製備    28
| II-2-2.  EDTA 溶液之製備    28
| II-3.  Nisin 及 EDTA之抗菌試驗    29
| II-3-1.  Nisin 對指標菌洋菜孔洞擴散法抗菌
| 活性之測試    29
| II-3-2.  不同培養溫度對六株革蘭氏陰性菌
| 之抗菌作用    29
| II-3-3.  不同起始 pH 值對六株革蘭氏陰性
| 菌之抗菌作用    30
| 四、結果與討論    31
| I.  六株革蘭氏陰性菌生長曲線之測定    31
| II.  洋菜孔洞擴散法實驗    31
| II-1.  Nisin 對指標菌的抗菌效果    31
| II-2.  Nisin 與 EDTA 對腸炎弧菌的抗菌效果    32
| III.  Nisin 與 EDTA 對革蘭氏陰性菌之抗菌效果    33
| III-1.  不同起始 pH 值之培養液在 26oC 下培養之
| 抗菌效果    33
| III-1-1.  對 V. parahaemolyticus isolates 1、5
| 、10、及 16 之抗菌效果    33
| III-1-2.  對 E. coli BCRC 11549、Sal.
| choleraesuis subsp. choleraesuis BCRC
| 10744 之抗菌效果    36
| III-2.  不同起始 pH 值之培養液在 37oC 下培養之
| 抗菌效果    37
| III-2-1.  對 V. parahaemolyticus 分離株1、5
| 、10、和 16之抗菌效果    37
| III-2-2.  對 E. coli BCRC 11549、Sal.
| choleraesuis subsp. choleraesuis BCRC
| 10744 之抗菌效果    40
| 五、結論    43
| 六、參考文獻    44
| 表 目 錄
|
| 表一、    細菌素做為食品防腐劑：可應用範圍的案例    63
| 表二、    各種細菌素名稱及其生產菌株    64
| 表三、    Nisin 在各國的使用狀況    65
| 表四、    控制組與實驗組經 50 □g mL-1 nisin 及 20 mM EDTA 在
| 不同起始 pH 值培養液中對六株革蘭氏陰性菌在 26oC
| 或 37oC 作用 0、1、3、6、9 或 12 hr 生菌量對數值
| 的差異    67
| 表五、    Nisin 及 EDTA 於 26oC及37oC 對 V. parahaemolyticus 分離株 1、5、10、和 16、以及 E. coli BCRC 11549 和 Sal.
| choleraesuis subsp. choleraesuis BCRC 10744 在 pH
| 4.0、5.0 及 6.0 之 TSB + 2.5% NaCl 及TSB 培養 12.0
| hr 後菌數之殘存率    69
|
|
| 圖 目 錄
|
| 圖一、    經轉譯後修飾作用所形成的 nisin 分子結構    70
| 圖二、    Nisin 抗菌作用之模式    71
| 圖三、    圖解革蘭氏陽性菌與革蘭氏陰性菌細胞套    72
| 圖四、    腸炎弧菌分離株 1、5、10、及 16 之生長曲線    73
| 圖五、    大腸桿菌 BCRC 11549 及沙門氏菌 BCRC 10744 之
| 生長曲線    74
| 圖六、    Nisin 對 Str. thermophilus BCRC 12268 的抗菌效果    75
| 圖七、    不同濃度 nisin 對 Str. thermophilus BCRC 12268 形
| 成的抑菌環    76
| 圖八、    不同起始 pH 值下 nisin 和 EDTA 對
| V. parahaemolyticus isolate 1 在 26oC 培養之抗菌
| 效果與 pH 值之變化    77
| 圖九、    不同起始 pH 值下 nisin 和 EDTA 對
| V. parahaemolyticus isolate 5 在 26oC 培養之抗菌
| 效果與 pH 值之變化    78
| 圖十、    不同起始 pH 值下 nisin 和 EDTA 對
| V. parahaemolyticus isolate 10 在 26oC 培養之抗菌
| 效果與 pH 值之變化    79
| 圖十一、    不同起始 pH 值下 nisin 和 EDTA 對
| V. parahaemolyticus isolate 16 在 26oC 培養之抗菌
| 效果與 pH 值之變化    80
| 圖十二、    不同起始 pH 值下 nisin 和 EDTA 對 E. coli
| BCRC 11549 在 26oC 培養之抗菌效果與 pH 值之
| 變化    81
| 圖十三、    不同起始 pH 值下 nisin 和 EDTA 對 Sal.
| choleraesuis subsp. choleraesuis BCRC 10744 在
| 26oC 培養之抗菌效果與 pH 值之變化    82
| 圖十四、    不同起始 pH 值下 nisin 和 EDTA 對
| V. parahaemolyticus isolate 1 在 37oC 培養之抗菌
| 效果與 pH 值之變化    83
| 圖十五、    不同起始 pH 值下 nisin 和 EDTA 對
| V. parahaemolyticus isolate 5 在 37oC 培養之抗菌
| 效果與 pH 值之變化    84
| 圖十六、    不同起始 pH 值下 nisin 和 EDTA 對
| V. parahaemolyticus isolate 10 在 37oC 培養之抗菌
| 效果與 pH 值之變化    85
| 圖十七、    不同起始 pH 值下 nisin 和 EDTA 對
| V. parahaemolyticus isolate 16 在 37oC 培養之抗菌
| 效果與 pH 值之變化    86
| 圖十八、    不同起始 pH 值下 nisin 和 EDTA 對 E. coli
| BCRC 11549 在 37oC 培養之抗菌效果與 pH 值之
| 變化    87
| 圖十九、    不同起始 pH 值下 nisin 和 EDTA 對 Sal.
| choleraesuis subsp. choleraesuis BCRC 10744 在
| 37oC 培養之抗菌效果與 pH 值之變化    88
|
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|
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