周正俊臺灣大學:食品科技研究所阮鳴嫣Juan, Ming-YenMing-YenJuan2010-05-112018-06-292010-05-112018-06-292008U0001-0708200818012900http://ntur.lib.ntu.edu.tw//handle/246246/182159Bacillus subtilis為日本具機能性傳統發酵食品－納豆之菌酛。納豆具有抗氧化之生理功效，並含有血管收縮素轉化酶抑制劑(angiotensin-converting enzyme inhibitors, ACEI) 可預防血壓上升。本研究利用B. subtilis BCRC 14715對黑豆進行固態發酵，探討不同溶劑 (水、80%甲醇、80%乙醇、80%丙酮)之發酵黑豆萃取物所呈現之抗氧化活性及總酚，總類黃酮與花青素之含量。抗氧化活性所檢測之項目包括DPPH自由基清除能力、還原力及亞鐵離子螯合能力。此外，本研究並探討發酵期間Bacillus spp.發酵黑豆萃取物對血管收縮素轉化酶Angiotensin I Converting Enzyme（ACE）之抑制活性，並比較不同Bacillus菌株發酵黑豆及不同溶劑萃取物對ACE之抑制作用。果顯示，黑豆無論是否發酵均呈現具有抗氧化活性，惟發酵黑豆萃取物所呈現之抗氧化活性皆顯著高於（p<0.05）未發酵黑豆者。此外發酵黑豆不同溶劑萃取物中總酚、總類黃酮、花青素含量也顯著高於（p<0.05）未發酵黑豆者，其中又以丙酮萃取物中之總酚、總類黃酮及花青素之含量最高。黑豆及發酵黑豆萃取物所呈現之抗氧化活性，因溶劑之不同而有所差異。甲醇萃取物對亞鐵離子呈現有最佳之螯合能力。而丙酮萃取物所呈現對DPPH自由基清除能力及還原力最佳。acillus spp.發酵黑豆之豆子或黏性物質萃取物，所呈現之ACE抑制活性皆隨發酵時間增長而提升，且因Bacillus菌株之不同而有差異，其中以Bacillus sp. CN11發酵黑豆者呈現之抑制活性最高。利用水、80% 乙醇萃取Bacillus spp. 發酵黑豆，在豆子部份以乙醇萃取物之ACE抑制能力較好，至於黏性物質部份則以水萃取物所呈現之ACE抑制活性顯著高於（p<0.05）乙醇萃取物。Natto, a traditional fermented food in Japan, is prepared by fermenting steamed soybean with Bacillus subtilis. It has been reported to exert antioxidant activities, and the inhibitory effect on angiotensin I converting enzyme (ACE) as well. In the present study, various solvent (water, 80% methanol, 80% ethanol, 80% acetone) extracts of B. subtilis BCRC 14715-fermented black soybean were examined for total phenolic, flavonoid, and anthocyanin contents as well as the antioxidant activities including 2,2-diphenyl-1-picrylhydrazyl (DPPH) radicals scavenging effects, reducing power, and Fe2+-chelating ability. Besides, effect of fermentation period and extraction solvent on ACE inhibitory activity of the Bacillus spp.-fermented black soybean extracts were also investigated.he results showed that the extract of the fermented black soybean exhibited significantly higher (p<0.05) total phenolic, flavonoid, anthocyanin contents and antioxidant activities than did the extract of the nonfermented black soybean. Among the various extracts examined, acetone extract showed the highest contents of total phenolic, flavonoid and anthocyanin. The methanol extract excerted the highest Fe2+-chelating ability, while the acetone extract showed the highest DPPH free radicals scavenging effects and reducing power.he inhibitory activity on ACE of bean and viscous material extracts of Bacillus spp.-fermented black soybean increased as the fermentation period was increased. Bacillus sp. CN11-fermented black soybean extract exhibited the highest ACE inhibitory activity. A higher ACE inhibitory activity was noted in bean extracted with 80% ethanol than with water. On the other hand, a significantly higher (p<0.05) ACE inhibitory activity was noted in the water extract of viscous material than the 80% ethanol extract.摘要-------------------------------------------------------------------------------------------- ibstract---------------------------------------------------------------------------------------- ii錄-------------------------------------------------------------------------------------------- iii目錄----------------------------------------------------------------------------------------- vii目錄----------------------------------------------------------------------------------------- viii、前言-------------------------------------------------------------------------------------- 1、文獻整理-------------------------------------------------------------------------------- 2、黑豆------------------------------------------------------------------------------------ 2.黑豆簡介----------------------------------------------------------------------------- 2.黑豆之抗氧化物質----------------------------------------------------------------- 2.1.總酚類化合物---------------------------------------------------------------- 3.2.總類黃酮---------------------------------------------------------------------- 3.3.花青素------------------------------------------------------------------------- 4、納豆------------------------------------------------------------------------------------ 4.納豆之成分與生理功能----------------------------------------------------------- 4.納豆之抗氧化活性----------------------------------------------------------------- 7、大豆發酵食品之抗氧化活性------------------------------------------------------ 8、萃取溶劑對萃取物抗氧化活性之影響------------------------------------------ 9、血管收縮素轉化酶 (Angiotensin I Converting Enzyme, ACE)-------------- 9. ACE之生化特性------------------------------------------------------------------- 9. ACE抑制劑------------------------------------------------------------------------- 10.大豆製品之ACE抑制活性------------------------------------------------------- 10、材料與方法----------------------------------------------------------------------------- 13、試驗架構------------------------------------------------------------------------------ 13、實驗材料------------------------------------------------------------------------------ 13.黑豆----------------------------------------------------------------------------------- 13.菌種----------------------------------------------------------------------------------- 13.培養基-------------------------------------------------------------------------------- 13.試驗藥品----------------------------------------------------------------------------- 13.儀器設備----------------------------------------------------------------------------- 15、試驗步驟與方法--------------------------------------------------------------------- 16.菌株保存----------------------------------------------------------------------------- 16.菌株冷凍保存----------------------------------------------------------------------- 16.菌株活化與接種菌液之製備----------------------------------------------------- 16.發酵黑豆之製備-------------------------------------------------------------------- 16、分析方法------------------------------------------------------------------------------ 17. 總酚、總類黃酮、花青素含量之測定------------------------------------------ 17.1總酚----------------------------------------------------------------------------- 17.2類黃酮-------------------------------------------------------------------------- 17.3花青素-------------------------------------------------------------------------- 17. 抗氧化活性之檢測--------------------------------------------------------------- 17.1 DPPH自由基清除能力----------------------------------------------------- 18.2還原力-------------------------------------------------------------------------- 19.3亞鐵離子螯合能力----------------------------------------------------------- 19. 血管收縮素轉化酶 (ACE) 抑制活性之檢測------------------------------- 20.1樣品之萃取------------------------------------------------------------------- 20.2分析方法---------------------------------------------------------------------- 20. 統計分析--------------------------------------------------------------------------- 21、結果與討論----------------------------------------------------------------------------- 22、黑豆與B. subtilis BCRC 14715發酵黑豆不同溶劑萃取物之萃出率------ 22、黑豆與B. subtilis BCRC 14715發酵黑豆不同溶劑萃取物中總酚、總類黃酮及花青素之含量--------------------------------------------------------------------- 2.總酚含量----------------------------------------------------------------------------- 24.總類黃酮含量----------------------------------------------------------------------- 24.花青素含量------------------------------------------------------------------------- 27、黑豆與B. subtilis BCRC 14715發酵黑豆不同溶劑萃取物之抗氧化活性------------------------------------------------------------------------------------------------ 30.DPPH自由基清除能力------------------------------------------------------------ 30.還原能力----------------------------------------------------------------------------- 33.亞鐵離子螯合能力----------------------------------------------------------------- 35、發酵期間Bacillus spp.發酵黑豆水萃取物對血管收縮素轉化酶之抑制活性--------------------------------------------------------------------------------------------- 40、Bacillus spp.發酵黑豆乙醇萃取物於發酵期間之血管收縮素轉化酶之抑制活性--------------------------------------------------------------------------------------- 44、不同Bacillus菌株發酵黑豆之水及80%乙醇萃取物對血管收縮素轉化酶之抑制作用------------------------------------------------------------------------------ 46、結論--------------------------------------------------------------------------------------- 50、參考文獻-------------------------------------------------------------------------------- 51錄一----------------------------------------------------------------------------------------- 63圖一、B. subtilis BCRC 14715發酵黑豆之豆子(A)黏性物質(B)水萃取物於發酵期間對血管收縮素轉化酶之抑制能力--------------------------------- 63圖二、Bacillus sp. CN11發酵黑豆之豆子(A)黏性物質(B)水萃取物於發酵期間對血管收縮素轉化酶之抑制能力------------------------------------------ 64圖三、B. subtilis BCRC 14715及Bacillus sp. CN11混合菌株發酵黑豆之豆子(A)黏性物質(B)水萃取物於發酵期間對血管收縮素轉化酶之抑制能力--------------------------------------------------------------------------------- 65圖四、B. subtilis BCRC 14715發酵黑豆之豆子(A)黏性物質(B)乙醇萃取物於發酵期間對血管收縮素轉化酶抑制能力--------------------------------- 66圖五、Bacillus sp. CN11發酵黑豆之豆子(A)黏性物質(B)80%乙醇萃取物於發酵期間對血管收縮素轉化酶之抑制能力--------------------------------- 67圖六、B. subtilis BCRC 14715及Bacillus sp. CN11混合菌株發酵黑豆之豆子(A)黏性物質(B)80%乙醇萃取物於發酵期間對血管收縮素轉化酶之抑制能力------------------------------------------------------------------------ 68錄二、台灣農業化學與食品科學期刊格式----------------------------------------------- 69application/pdf818600 bytesapplication/pdfen-USBacillus subtilis黑豆萃取溶劑抗氧化活性血管收縮素轉化酶black soybeanextraction solventantioxidant activityangiotensin I converting enzyme (ACE)thesishttp://ntur.lib.ntu.edu.tw/bitstream/246246/182159/1/ntu-97-R95641027-1.pdf