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研究生:吳俊億
研究生(外文):Jiun-Yi Wu
論文名稱:利用根黴菌開發機能性醱酵黑豆食品之研究
論文名稱(外文):Development of Functional Fermented Foods from Black Soybean by Rhizopus spp.
指導教授:劉文雄劉文雄引用關係
指導教授(外文):Wen-Hsiung Liu
學位類別:碩士
校院名稱:國立臺灣大學
系所名稱:微生物與生化學研究所
學門:生命科學學門
學類:微生物學類
論文種類:學術論文
論文出版年:2004
畢業學年度:92
語文別:中文
論文頁數:90
中文關鍵詞:黑豆根黴菌異黃酮固態醱酵
外文關鍵詞:isoflavonesblack soybeansolid state fermentationRhizopus spp.
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本研究利用省產黑豆為原料以根黴菌之醱酵法生產機能性醱酵黑豆食品,並對醱酵黑豆產品進行弁鄔岍侅�及弁鄔呇角壑尷R,以期確立機能性醱酵黑豆產品之醱酵生產體系,開拓省產黑豆在健康食品上之用途為目的。
探討以高效能液相層析法有效分離四種異黃酮標準品之條件,得知以65℃萃取2小時可得最高量異黃酮含量及最佳DPPH自由基之清除效應。以天貝 (tempeh)模式,檢討根黴菌對黑豆台南三號進行固態醱酵期間代謝產物之變化,得知三株根黴菌檢測菌均可將未脫皮黑豆及脫皮黑豆中具有醣基之異黃酮 (daidzin及genistin) 轉換成不具醣基之異黃酮 (daidzein及genistein)。此轉換反應與根黴菌所分泌之β-葡萄醣苷酶之活性有正相關性,依據異黃酮轉換效率、 自由基之清除效應及醱酵時間等因子,選擇以R. oligosporus NTU-5醱酵脫皮黑豆台南三號進行抗氧化物質之純化與分離。以矽膠管柱層析法進行區分獲得以正己烷:乙酸乙酯(1/2, v/v)沖提之區分 3具有最大之抗氧化活性,以薄層層析法及半製備式高效能液相層析法分離出不具醣基之異黃酮daidzein、genistein及具抗氧化活性未知化合物SSFE-PA。Daidzein及genistein之回收率分別為94.6% 及58.6% 。未知化合物SSFE-PA對DPPH自由基之有效半抑制濃度(IC50)值為9.8 μg/mL,以液相層析質譜儀分析其分子量為287。以R. oligosporus NTU-5對黑豆台南三號粉末進行液態醱酵,由醱酵液之分析結果得知黑豆具有醣基之異黃酮亦可被轉換成不具醣基之異黃酮。


The purpose of this study is to develop a fermentation process for production of functional fermented foods from domestic black soybean by Rhizopus spp. The functional components of fermented black soybean and its physiological function will also be investigated. These results can provide information for establishment of a fermentation process for production of functional fermented black soybean.
A fast, precise method is presented for the efficient separation of four isoflavones by high performance liquid chromatorgraphy. The content of isoflavones and 1,1-diphenyl-2-dipicrylhydrazyl (DPPH) radical scavenging test reached to the best by extracted at 65℃ for 2 h. Studies on solid state fermentation by tempeh-like process and the changes of metabolites in black soybeans Tainan NO.3 were investigated by using Rhizopus spp. It was found that the conversion of isoflavones glycosides (daidzin ; genistin) in black soybean to isoflavones aglycones (daidzein ; genistein) was contributed to β-glucosidase activity in fermented black soybeans produced by Rhizopus spp. According to the content of isoflavones aglycones formation, scavenging effect and cultivation time, fermented dehulled black soybeans with R. oligosporus NTU-5 was selected to further isolation and purification of antioxidant. A fraction 3 had the highest scavenging effect was isolated from silica gel column chromatography, eluting with ethyl acetate/n-hexane (1/2, v/v). Then, the fraction 3 was analyzed and purified by TLC and semi-preparative HPLC. Daidzein, genistein and an unknown antioxidative compound SSFE-PA were isolated. The recovery of daidzein and genistein were 94.6% and 58.6%, respectively. The IC50 value of SSFE-PA for DPPH free radicals was 9.8 μg/mL. The molecular weight of SSFE-PA was 287 dalton analyzed by LC/MS. A submerged fermentation of black soybean Tainan NO.3 was also investigated by R. oligosporus NTU-5. It was found that the isoflavones glycosides in black soybean Tainan NO.3 was converted to isoflavones aglycones via the submerged fermentation of R. oligosporus NTU-5.


中文摘要 ......................................... Ⅰ
英文摘要 ......................................... Ⅱ
目錄 ............................................. Ⅳ
表次 ............................................. Ⅴ
圖次 .............................................. Ⅵ
縮寫表 ............................................ Ⅷ

第一章 緒言 ...................................... 1
第一節 黑豆 ..................................... 1
第二節 根黴菌簡介 ............................... 3
第三節 自由基與健康 ............................. 4
第四節 植物性化學物質 ........................... 8第五節 醱酵食品 ................................. 18
第六節 研究動機與目的 ........................... 23

第二章 材料與方法 ............................... 26
第一節 材料 ..................................... 26
第二節�分析方法 ................................. 27
第三節�實驗方法 ................................. 35
第四節�藥品 ..................................... 38
第五節�儀器 ..................................... 38

第三章 結果 ..................................... 41
第一節 異黃酮萃取之條件 ......................... 41
第二節 篩選最適化固態醱酵之根黴菌 ............... 41
第三節 固態醱酵條件之建立 ....................... 47第四節 抗氧化物之純化與分離 ..................... 54第五節 液態醱酵之探討 ........................... 65

第四章 討論 ..................................... 75
第五章 結論 ..................................... 83
第六章 參考文獻 .................................. 85


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