周正俊臺灣大學：食品科技研究所王妍如Wang, Yen-JuYen-JuWang2007-11-272018-06-292007-11-272018-06-292007http://ntur.lib.ntu.edu.tw//handle/246246/56350本研究以Aspergillus awamori作為菌酛製備黑豆麴，探討在不同溫度（40℃、60℃、80℃或100℃）下加熱30分鐘，在不同包裝條件下（乾燥劑＋脫氧劑、乾燥劑、脫氧劑與未添加任何食品保存劑）於4℃或25℃中儲存120天後黑豆麴對4-nitroquinoline-N-oxide及Benzo[a]pyrene抗致突變性之影響。 結果顯示，黑豆麴在40℃加熱30分鐘後，其總酚類化合物即顯著性地下降（p＜0.05），加熱溫度提高至80℃以上時亦造成其花青素含量顯著性地（p＜0.05）降低。當溫度提高至80℃或以上加熱30分鐘後，會顯著地（p＜0.05）降低其甲醇萃取物對4-NQO與B[a]P之抗致突變能力。萃取物抗致突變活性降低之程度因所測試菌株及致突變劑之不同而有所差異。 儲存溫度與包裝方式會影響黑豆麴中總酚類化合物、花青素之含量及其甲醇萃取物所呈現之抗致突變活性。隨著儲存時間之延長，花青素與總酚類化合物之含量以及抗致突變活性均呈降低之現象。一般而言，黑豆麴在4℃下儲存所保有之總酚類化合物、花青素含量及抗致突變活性高於在25℃下儲存者。在所測試之包裝方式中，黑豆麴儲存時同時添加脫氧劑與乾燥劑最能保有總酚類化合物、花青素之含量及其抗致突變活性。未添加乾燥劑與脫氧劑者，所呈現之保存效果最差。In the present study, black soybean koji were first prepared with Aspergillus awamori. They were then subjected to heating at various temperatures (40, 60, 80 and 100℃) for 30 min or storage at either 4 or 25℃ under different packaging conditions (dessicant and deoxidant, dessicant, deoxidant, without dessicant and deoxidant) for 120 days. It was found that the anthocyanin content decreased significantly（p＜0.05） in black soybean koji after heating at 80℃ or higher temperature. While the total phenolic content reduced significantly （p＜0.05） after heating at 40℃ or higher. Heating at 80℃ or higher for 30 min also resulted in a significant reduction（p＜0.05）in the antimutagenicity of the methanol extract of black soybean koji against 4-nitroquinoline N-oxide and Benzo[a]pyrene. However, extent of reduction in antimutagenicity varied with the test strains of S. typhimurium and the kind of mutagens examined. Regardless of storage temperature and packaging conditions, content of total phenolic, anthocyanin and antimutagenic activity of the black soybean koji decreased as the storage period was extended. Higher retention of total phenolic, anthocyanins and the antimutagenic activity was noted in black soybean koji held at 4℃ than at 25℃. Among the various packaging conditions examined, black soybean koji stored with dessicant and deoxidant exhibited the highest retention of total phenolics, anthocyanin and antimutagenic activity during storage. While the poorest retention was noted with black soybean koji stored without dessicant and deoxident.中文摘要--------------------------------------------------i 英文摘要-------------------------------------------------ii 目錄----------------------------------------------------iii 圖目錄----------------------------------------------------v 表目錄---------------------------------------------------vi 壹、前言--------------------------------------------------1 貳、文獻回顧----------------------------------------------2 一、黑豆------------------------------------------------2 1. 黑豆簡介------------------------------------------2 2. 黑豆之機能性--------------------------------------2 2.1酚類化合物----------------------------------------3 2.2花青素--------------------------------------------3 2.3異黃酮--------------------------------------------4 二、發酵食品之抗致突變性--------------------------------4 1. 微生物之抗致突變性--------------------------------5 2. 微生物發酵食品之抗致突變性------------------------5 三、熱處理對抗致突變活性之影響--------------------------6 四、儲存條件對抗致突變性之影響--------------------------6 參、材料與方法--------------------------------------------8 一、材料------------------------------------------------8 1. 菌種----------------------------------------------8 2. 黑豆----------------------------------------------8 3. 培養基--------------------------------------------8 4. 試驗藥品------------------------------------------8 5. 儀器設備------------------------------------------9 二、樣品製備-------------------------------------------10 1. 菌株之保存與活化---------------------------------10 2. 冷凍保存-----------------------------------------10 3. 菌株之活化與孢子液製備---------------------------10 4. S. typhimurium突變株之保存活化-------------------10 5. 黑豆麴之製備-------------------------------------10 6. 黑豆麴甲醇萃取物之製備---------------------------10 7. 腸胃液體之組成份---------------------------------11 三、分析方法-------------------------------------------11 1. S. typhimurium TA98與TA100菌株基因型態之確認-----11 1.1組胺酸要求性-------------------------------------11 1.2 rfa突變之測試-----------------------------------12 1.3 uvrB突變之測試----------------------------------12 1.4 R-factor之測試----------------------------------12 2. 抗致突變性試驗-----------------------------------13 2.1 毒性試驗----------------------------------------13 2.2 致突變性試驗------------------------------------13 2.3 抗致突變性試驗----------------------------------14 3. 熱穩定性對黑豆麴抗致突變性之探討-----------------14 4. 胃液與腸液處理對黑豆麴抗致突變性之影響-----------15 4.1 胃液處理----------------------------------------15 4.2 腸液處理----------------------------------------15 5. 儲存條件對黑豆麴抗致突變之試驗-------------------15 6. 總酚類化合物含量測定-----------------------------15 7. 黑豆麴萃取液之總花青素含量測定-------------------15 8. 統計分析-----------------------------------------16 肆、結果與討論-------------------------------------------17 一、S. typhimurium TA98及TA100試驗菌株基因型態之確認---17 1.組胺酸需求測定------------------------------------17 2. rfa突變之測試------------------------------------17 3. uvrB突變之測試-----------------------------------17 4. R-factor之測試-----------------------------------17 二、加熱處理對黑豆麴抗致突變性之影響-------------------19 1. 加熱處理黑豆麴對S. typhimurium TA98及TA100之毒性-19 2. 加熱處理黑豆麴對S. typhimurium TA98及TA100之致突變 性-----------------------------------------------19 3. 加熱處理黑豆麴對S. typhimurium TA98及TA100之抗致突 變性---------------------------------------------21 4. 加熱處理對黑豆麴中總酚類化合物與花青素含量之影響-26 三、模擬腸胃道對黑豆麴抗致突變性之影響-----------------32 1. 黑豆麴萃取物在模擬胃液與腸液處理對S. typhimurium TA98與TA100之毒性--------------------------------32 2. 黑豆麴萃取物在模擬胃液與腸液處理對S. typhimurium TA98與TA100之致突變性----------------------------32 3. 黑豆麴萃取物在模擬胃液處理對S. typhimurium TA98與 TA100抗致突變性之影響----------------------------32 4. 黑豆麴萃取物在模擬腸液處理對S. typhimurium TA98與 TA100抗致突變性之影響----------------------------37 四、儲存條件對抗致突變性之影響-------------------------40 1. 不同儲存條件下之黑豆麴對S. typhimurium TA98與TA100 之毒性-------------------------------------------40 2. 不同儲存條件下之黑豆麴對S. typhimurium TA98與TA100 之致突變性---------------------------------------40 3. 不同儲存條件下之黑豆麴對S. typhimurium TA98與TA100 之抗致突變性-------------------------------------40 4. 儲存條件對黑豆麴中總酚化合物與花青素含量之影響---50 伍、結論-------------------------------------------------53 陸、參考文獻---------------------------------------------54 柒、附錄-------------------------------------------------65 附表一、不同包裝條件下黑豆麴於4℃儲存120天後其萃取物對4-NQO在 S. typhimuriumnTA98與TA100中之抗致突變性--------------65 附表二、不同包裝條件下黑豆麴25℃儲存120天後其萃取物對4-NQO在 S. typhimuriumn TA98與TA100中之抗致突變性-------------66 附表三、不同包裝條件下黑豆麴4℃儲存120天後其萃取物對B[a]P在 S. typhimuriumn TA98與TA100中之抗致突變性-------------67 附表四、不同包裝條件下黑豆麴25℃儲存120天後其萃取物對B[a]P在 S. typhimuriumn TA98與TA100中之抗致突變性-------------68494967 bytesapplication/pdfen-US黑豆麴抗致突變性加熱處理包裝條件花青素總酚Black soybean kojiAntimutagenicityHeat treatmentPackaging conditionsanthocyanintotal phenolicthesishttp://ntur.lib.ntu.edu.tw/bitstream/246246/56350/1/ntu-96-R94641017-1.pdf