周正俊臺灣大學：食品科技研究所謝孟荔Hsieh, Meng-LiMeng-LiHsieh2007-11-272018-06-292007-11-272018-06-292004http://ntur.lib.ntu.edu.tw//handle/246246/56372本研究首先以乳酸菌 (S. thermophilus, L. acidophilus) 與雙叉桿菌 (B. longum, B. infantis) 單獨或混合菌株製備發酵豆奶，然後利用Ames test 測試發酵豆奶對4-nitroquinoline-N-oxide(4-NQO)及3,2’-dimethyl-4-aminobiphenyl(DMAB) 所誘發突變之抗致突變活性。 實驗結果顯示經發酵處理之豆奶能顯著(p<0.05)效提高豆奶之抗致突變性；然而，發酵豆奶所呈現之抗致突變活性因發酵菌酛之不同而有所差異。在所試之發酵豆奶中以S. thermophilus、B. infantis混合發酵之發酵豆奶對4-NQO及DMAB具有最高之抗致突變活性，分別達85.07 %及85.78 %。在發酵過程中，發酵豆奶之抗致突變活性隨著發酵時間之延長而上升，發酵豆奶對4-NQO抗致突變活性之上升與豆奶中乳酸菌和雙叉桿菌總菌數之增加相一致；惟發酵豆奶對DMAB誘發突變抑制率之上升情形與菌量之增加並不一致。 添加菌酛但未進行發酵之豆奶之抗致突變活性僅與豆奶相當，均低於發酵豆奶組；此顯示，發酵豆奶提高之抗致突變活性主要乃於發酵過程中形成。 發酵豆奶並不具生物抗致突變性或去致突變性。其所呈現之抗致突變活性主要源自於Blocking effect之效果。In this study, soymilk was first fermented with lactic acid bacteria (Streptococcun thermophilus, Lactobacillus acidophilus) and bifidobacteria (Bifidobacterium infantis, Blfidobacterium infantis) alone or simultaneously. The antimutagenicity of soymilk and fermented soymilk against 4-nitroquinoline-N-oxide (4-NQO) and 3,2’-dimethyl-4-amino-biphenyl (DMAB) was then investigated with Ames test. Results revealed that fermentation significantly (p<0.05) increased the antimutagenicity activity of soymilk. The antimutagenicity of fermented soymilk varied with the starter used. Soymilk fermented with both S. themophilus and B. infantis simultaneously exhibited an antimutagenicity of 85.07% and 85.78%, respectively, against 4-NQO and DMAB. They were the highest among the various fermented soymilk products tested. It was also noted that the antimutagenic activity of fermented soymilk against 4-NQO was increased as the cultivation time expended and coincided with the increase of the total viable population of lactic acid bacteria and bifidobacteria. While,this phenomenon was not observed when antimutagenic activity of fermented soymilk was tested against DMAB. Antimutagenic activity of the soymilk added with starter was found to be similar to that of soymilk, while lower than the fermented soymilk. This indicated that the increased antimutagenic activity observed with fermented soymilk was related to the fermentation process. Fermented soymilk exhibited no inhibitory effects toward 4-NQO or DMAB in the bio-antimutagenic assay. Blocking effect was the main mechanism which led to the observed antimutagenic effect against the mutagens tested.中文摘要 I 英文摘要 Ⅱ 目錄 Ⅲ 圖次 Ⅵ 表次 Ⅶ 壹、前言 1 貳、文獻回顧 3 一、 黃豆及其製品 3 1. 黃豆的化學組成 3 2. 黃豆的保健機能 5 2.1 異黃酮 5 2.2 皂素 6 2.3 維生素E 6 3. 豆奶及發酵豆奶 7 3.1 豆奶 7 3.2 發酵豆奶 10 3.2.1 發酵豆奶之優點 10 二、 益生菌 12 1. 益生菌之定義及發展歷史 12 2. 益生菌與人体腸道健康之關係 13 3. 益生菌應具備之條件 14 4. 益生菌的生理功能 14 4.1 維持正常之微生物菌相 14 4.2 提高生物利用率及合成營養素 14 4.3 促進胃腸道健康 16 4.4 改善乳糖不耐症 16 4.5 活化免疫系統 17 4.6 抗腫瘤 17 4.7 降低過敏症的發生 18 4.8 降低血液中膽固醇含量 18 三、 膳食與癌症 19 1. 膳食中的抗致突變物 20 2. 益生菌及其發酵製品之抗致突變及抗癌活性 22 四、 抗致突變檢測方法及其機制 24 1. 安氏實驗法(Amest test) 24 2. 抗致突變物/抗致癌物之作用機制 25 參、材料與方法 33 一、 實驗材料 33 1. 試驗菌種 33 2. 黃豆 33 3. 藥品及培養基 33 4. 儀器及器材 34 二、實驗方法 35 1. 菌株之活化與接種源之製備 35 1.1乳酸菌及雙叉桿菌之保存活化與製備 35 1.2 Salmonella typhimurium TA100之保存活化 35 2. Salmonella typhimurium TA100菌株基因形態之確認 35 2.1 組胺酸要求性 36 2.2 rfa突變之測試 37 2.3 uvrB突變之測試 37 2.4 R-factor之測試 37 3. 豆奶基質及發酵豆奶之製備 38 4. 抗致突變性試驗 38 4.1 毒性試驗 38 4.2 致突變試驗 39 4.3 抗致突變性試驗 39 5. 不同發酵菌酛發酵豆奶之抗致突變性 40 6. 未發酵豆奶添加菌酛與利用菌酛發酵豆奶抗致突變性之比較 41 7. 發酵豆奶抗致突變可能機制之探討 41 7.1 生物抗致突變性之分析 41 7.2 去致突變性試驗 42 7.2.1 發酵豆奶對致突變劑之影響 42 7.2.2 發酵豆奶對S9 mix之影響 42 7.2.3 發酵豆奶對DMAB代謝活化物之影響 42 7.3 Blocking Effect試驗 42 8. 乳酸菌與雙叉桿菌菌數之測定 43 9. 統計分析 43 肆、結果與討論 44 一、 Salmonella typhimurium TA100試驗菌株基因形態之確認 44 二、 乳酸菌及雙叉桿菌發酵豆奶對S. typhimurium TA100之細胞毒性試驗、致突變性試驗及抗致突變試驗 46 1. 毒性試驗 46 2. 致突變試驗 48 3. 乳酸菌及雙叉桿菌發酵豆奶之抗致突變性 50 三、 豆奶發酵過程中菌數、pH值及抗致突變性之變化 52 四、 未發酵豆奶添加菌酛與利用菌酛發酵豆奶抗致突變性之比較 54 五、 發酵豆奶抗致突變之可能機制 59 1. 生物抗致突變性 (bioantimutagenesis) 59 2. 去致突變性 (desmutagenesis) 61 3. Blocking Effect 65 4. 發酵豆奶抗致突之可能機制—結論 69 伍、結論 70 陸、參考文獻 711239156 bytesapplication/pdfen-US乳酸菌抗致突變益生菌發酵豆奶雙叉桿菌probioticbifidobacterialactic acid bacteriafermented soymilkantimutagenicitythesishttp://ntur.lib.ntu.edu.tw/bitstream/246246/56372/1/ntu-93-R91641007-1.pdf