探討以 Eurotium cristatum 發酵轉換清豆漿與 藍莓汁之異黃酮糖苷與葡萄糖苷酶抑制能力

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傅啟堯(Chi-Yao Fu) 查詢紙本館藏

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化學工程與材料工程學系

論文名稱

探討以 Eurotium cristatum 發酵轉換清豆漿與藍莓汁之異黃酮糖苷與葡萄糖苷酶抑制能力
(Evaluation of isoflavone glucosides bioconversion and a-glucosidase inhibition ability in soymilk fermented with Cyanococcus’s juice by Eurotium cristatum)

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至系統瀏覽論文 (2026-8-31以後開放)

摘要(中)

富含蛋白質的大豆產品，能提供人體營養所需之胺基酸及礦物質，大豆不僅可以降低血清中總膽固醇(Total Cholesterol)、低密度脂蛋白(Low-density lipoproteins) 以及甘油三酸脂(Triglyceride)的濃度，還能提升高密度脂蛋白(High-density lipoproteins)。此外，發酵後的大豆製品亦被證明可有效減輕糖尿病、血壓、心臟疾病以及和癌症相關的問題。經過發酵後的大豆製品，營養價值比未發酵的大豆還來的高，因此，本論文主要利用金花菌發酵黃豆，並探討其組成以及營養價值。
本實驗以市售之黃豆作為原料，將其浸泡12小時後，榨汁製成清豆漿。利用自茶葉分離出的金花菌株(Eurotium cristatum)接種至清豆漿與豆渣發酵三天，並將溫度控制於25℃、30℃、35℃、40℃，搖瓶轉速設定為125rpm，分析其酵素抑制能力、抗氧化活性、總多酚含量以及異黃酮苷元轉換率。並發現於30℃、35℃下發酵可得最佳酵素抑制能力與產物濃度及最大菌重濃度。
此外，因藍莓含有豐富的抗氧化物、抗癌物質、多酚類，可以降低心血管和中風等疾病。藍莓中所含之花青素能調節胰島素敏感度，以降低血糖水平，有助於預防及改善第2型糖尿病，還有抑制體內發炎的效果。故將添加藍莓之清豆漿做為實驗對照組，比較其酵素抑制之效果。實驗結果發現於35℃下添加10%豆漿的發酵液有最佳酵素抑制能力及抗氧化力。

摘要(英)

Soy products are rich in protein and have many health benefits. Some researchers have pointed out that soybeans can reduce the serum concentrations of total cholesterol, low-density lipoproteins, and triglycerides in the serum. In addition, fermented soy products have also been proven to be effective in reducing diabetes mellitus, blood pressure, heart disease, and cancer-related problems. Fermented soybean products have higher nutritional value than unfermented soybeans. Therefore, this work is about using Eurotium cristatum to ferment soymilk, and discusses its composition and nutritional value.
In this study, soybeans was raw materials. It was soaked for 12 hours, and then juiced into clear soy milk. Eurotium cristatum was isolated from tea and inoculated to soy milk and soy residue for three days of fermentation, and the temperature was controlled at 25℃、30℃、35℃、40℃ respectively, the shaker’s speed was set at 125rpm, and the samples’ enzyme inhibition ability, antioxidant activity, total polyphenol content and isoflavone bioconversion rate were analyzed. It was found that the optimal enzyme inhibition ability and product concentration and maximum bacterial weight concentration were obtained from fermentation at 30℃ and 35℃.
In addition, blueberries are rich in antioxidants, anti-cancer elements, polyphenols, which can reduce cardiovascular and stroke and other diseases. The anthocyanins contained in blueberries regulate insulin sensitivity to lower blood sugar levels, help prevent and improve type 2 diabetes, and inhibit inflammation in the body. Therefore, the blueberry was added to clear soy milk as an experimental control group, comparing the effect of its enzyme inhibition. The results showed that the fermentation liquid with 10% soy milk at 35℃ had highest enzyme inhibition and antioxidant ability.

關鍵字(中)

★ 發酵
★ 豆漿
★ 金花菌
★ 藍莓
★ 大豆醣苷
★ 花色素苷

關鍵字(英)

★ fermentation
★ soymilk
★ Eurotium cristatum
★ Cyanococcus
★ isoflavone glucosides
★ anthocyanins

論文目次

目錄
摘要 I
Abstract II
致謝 IV
目錄 V
圖目錄 VIII
表目錄 XIII
第一章緒論 1
1.1 研究動機 1
1.2 研究目的 2
第二章文獻回顧 3
2.1 黃豆介紹 3
2.1.1 成分組成 4
2.1.2 黃豆功能 10
2.2 藍莓介紹 11
2.2.1 藍莓功能 11
2.3 金花菌介紹(Eurotium cristatum) 12
2.3.1 金花菌功能 13
2.4糖尿病介紹(Diabetes) 14
2.4.1 糖尿病的成因和類別 14
2.4.2第2型糖尿病的治療方法 15
第三章材料與方法 16
3.1實驗規劃 16
3.2實驗藥品 17
3.3儀器設備 18
3.4實驗方法 19
3.4.1 分離菌種 19
3.4.2 培養基處理 21
3.5分析方法 22
3.5.1 菌重測定 22
3.5.2 異黃酮類分析 22
3.5.3 α-澱粉酶抑制能力測定(α-amylase inhibition ability) 27
3.5.4 α-葡萄糖苷酶抑制能力測定(α-glucosidase activity inhibition ability) 30
3.5.5 DPPH自由基清除能力(DPPH scavenging activity) 31
3.5.6 總多酚類化合物含量測定(Total phenolic compounds) 32
3.5.7 花色素苷含量分析(Total anthocyanins content) 34
3.5.8 黃酮類含量分析(Flavonoids) 35
3.5.9 β-葡萄糖苷酶活性測試(β-Glucosidase activity assay) 36
第四章結果與討論 37
4.1發酵溫度對發酵液抗氧化活性及總多酚含量的影響 37
4.2不同溫度對以冠突散囊菌生產大豆異黃酮發酵動力曲線之影響 43
4.3不同溫度對豆漿發酵液及豆渣萃取液澱粉酶抑制活性的影響 58
4.4不同溫度對豆漿發酵液及豆渣萃取液葡萄糖苷酶抑制活性的影響 61
4.5 30℃下添加稀釋豆漿對藍莓發酵液的影響 65
4.6 35℃下添加稀釋豆漿對藍莓發酵液的影響 76
第五章結論 87
第六章參考資料 88

圖目錄
圖 1、大豆發酵前後異黃酮類HPLC圖 6
圖 2、染料木黃酮和黃豆苷元與17-β-雌二醇之結構相似性 7
圖 3、β-葡萄糖苷酶對異黃酮糖苷的反應機制圖 8
圖 4、實驗流程圖 16
圖 5、Eurotium cristatum 純化後生長於PDA固態盤外觀 20
圖 6、各產物之HPLC圖譜 24
圖 7、染料木苷標準品濃度之檢量線 25
圖 8、大豆醣苷標準品濃度之檢量線 25
圖 9、染料木黃酮標準品濃度之檢量線 26
圖 10、大豆苷元標準品濃度之檢量線 26
圖 11、沒食子酸標準品檢量線 33
圖 12、蘆丁標準品檢量線 35
圖 13、發酵時間對於25℃豆漿發酵液之抗氧化活性變化 38
圖 14、發酵時間對於30℃豆漿發酵液之抗氧化活性變化 39
圖 15、發酵時間對於35℃豆漿發酵液之抗氧化活性變化 39
圖 16、發酵時間對於40℃豆漿發酵液之抗氧化活性變化 40
圖 17、發酵時間對於25℃豆渣萃取液之抗氧化活性變化 40
圖 18、發酵時間對於30℃豆渣萃取液之抗氧化活性變化 41
圖 19、發酵時間對於35℃豆渣萃取液之抗氧化活性變化 41
圖 20、發酵時間對於40℃豆渣萃取液之抗氧化活性變化 42
圖 21、發酵時間對25℃豆漿發酵液PH值與β-葡萄糖苷酶活性變化 43
圖 22、發酵時間對30℃豆漿發酵液PH值與β-葡萄糖苷酶活性變化 43
圖 23、發酵時間對35℃豆漿發酵液PH值與β-葡萄糖苷酶活性變化 44
圖 24、發酵時間對40℃豆漿發酵液PH值與β-葡萄糖苷酶活性變化 44
圖 25、發酵時間對25℃豆渣萃取液PH值與β-葡萄糖苷酶活性變化 45
圖 26、發酵時間對30℃豆渣萃取液PH值與β-葡萄糖苷酶活性變化 45
圖 27、發酵時間對35℃豆渣萃取液PH值與β-葡萄糖苷酶活性變化 46
圖 28、發酵時間對40℃豆渣萃取液PH值與β-葡萄糖苷酶活性變化 46
圖 29、Eurotium cristatum在25℃對豆漿發酵液中大豆苷元之發酵曲線 49
圖 30、Eurotium cristatum在25℃對豆漿發酵液中染料木黃酮之發酵曲線 49
圖 31、Eurotium cristatum在30℃對豆漿發酵液中大豆苷元之發酵曲線 50
圖 32、Eurotium cristatum在30℃對豆漿發酵液中染料木黃酮之發酵曲線 50
圖 33、Eurotium cristatum在35℃對豆漿發酵液中大豆苷元之發酵曲線 51
圖 34、Eurotium cristatum在35℃對豆漿發酵液中染料木黃酮之發酵曲線 51
圖 35、Eurotium cristatum在40℃對豆漿發酵液中大豆苷元之發酵曲線 52
圖 36、 Eurotium cristatum在40℃對豆漿發酵液中染料木黃酮之發酵曲線 52
圖 37、Eurotium cristatum在25℃對豆渣中大豆苷元之發酵曲線 54
圖 38、Eurotium cristatum在25℃對豆渣中染料木黃酮之發酵曲線 54
圖 39、Eurotium cristatum在30℃對豆渣中大豆苷元之發酵曲線 55
圖 40、Eurotium cristatum在30℃對豆渣中染料木黃酮之發酵曲線 55
圖 41、Eurotium cristatum在35℃對豆渣中大豆苷元之發酵曲線 56
圖 42、Eurotium cristatum在35℃對豆渣中染料木黃酮之發酵曲線 56
圖 43、Eurotium cristatum在40℃對豆渣中大豆苷元之發酵曲線 57
圖 44、Eurotium cristatum在40℃對豆渣中染料木黃酮之發酵曲線 57
圖 45、25℃豆漿發酵液及豆渣萃取液澱粉酶抑制活性變化 59
圖 46、30℃豆漿發酵液及豆渣萃取液澱粉酶抑制活性變化 59
圖 47、35℃豆漿發酵液及豆渣萃取液澱粉酶抑制活性變化 60
圖 48、40℃豆漿發酵液及豆渣萃取液澱粉酶抑制活性變化 60
圖 49、25℃豆漿發酵液及豆渣萃取液葡萄糖苷酶抑制活性變化 62
圖 50、30℃豆漿發酵液及豆渣萃取液葡萄糖苷酶抑制活性變化 62
圖 51、35℃豆漿發酵液及豆渣萃取液葡萄糖苷酶抑制活性變化 63
圖 52、40℃豆漿發酵液及豆渣萃取液葡萄糖苷酶抑制活性變化 63
圖 53、30℃不同濃度豆漿的抗氧化活性 66
圖 54、30℃藍莓發酵液添加稀釋豆漿對抗氧化活性變化 66
圖 55、30℃藍莓發酵液添加稀釋豆漿對花色素苷含量變化 67
圖 56、30℃藍莓發酵液添加稀釋豆漿對水溶性黃酮類含量變化 67
圖 58、30℃藍莓發酵液添加稀釋豆漿對葡萄糖苷酶抑制活性變化 69
圖 59、30℃藍莓發酵液添加稀釋豆漿對菌種生長曲線變化 70
圖 60、30℃藍莓發酵液於不同豆漿濃度下對最大菌重圖 70
圖 61、30℃藍莓發酵液之PH值與β-葡萄糖苷酶活性變化 71
圖 62、30℃藍莓發酵液添加5%豆漿之PH值與β-葡萄糖苷酶活性變化 72
圖 63、30℃藍莓發酵液添加10%豆漿PH值與β-葡萄糖苷酶活性變化 72
圖 64、30℃藍莓發酵液添加5%豆漿中大豆苷元之發酵曲線 74
圖 65、30℃藍莓發酵液添加5%豆漿中染料木黃酮之發酵曲線 74
圖 66、30℃藍莓發酵液添加10%豆漿中大豆苷元之發酵曲線 75
圖 67、30℃藍莓發酵液添加10%豆漿中染料木黃酮之發酵曲線 75
圖 68、35℃不同濃度豆漿的抗氧化活性 77
圖 69、35℃藍莓發酵液添加稀釋豆漿對抗氧化活性變化 77
圖 70、35℃藍莓發酵液添加稀釋豆漿對花色素苷含量變化 78
圖 71、35℃藍莓發酵液添加稀釋豆漿對水溶性黃酮類含量變化 78
圖 72、35℃藍莓發酵液添加稀釋豆漿對澱粉酶抑制活性變化 80
圖 73、35℃藍莓發酵液添加稀釋豆漿對葡萄糖苷酶抑制活性變化 80
圖 74、35℃藍莓發酵液添加稀釋豆漿對菌種生長曲線變化 81
圖 75、35℃藍莓發酵液於不同豆漿濃度下對最大菌重圖 81
圖 76、35℃藍莓發酵液之PH值與β-葡萄糖苷酶活性變化 82
圖 77、35℃藍莓發酵液添加5%豆漿之PH值與β-葡萄糖苷酶活性化 83
圖 78、35℃藍莓發酵液添加10%豆漿PH值與β-葡萄糖苷酶活性變化 83
圖 79、35℃藍莓發酵液添加5%豆漿中大豆苷元之發酵曲線 85
圖 80、35℃藍莓發酵液添加5%豆漿中染料木黃酮之發酵曲線 85
圖 81、35℃藍莓發酵液添加10%豆漿中大豆苷元之發酵曲線 86
圖 82、35℃藍莓發酵液添加10%豆漿中染料木黃酮之發酵曲線 86

表目錄
表 1、大豆隨發酵時間的異黃酮類成分變化 7
表 2、藥品清單 17
表 3、實驗設備清單 18
表 4、HPLC移動相設定參數 23
表 5、不同溫度下豆漿搖瓶發酵參數值 48
表 6、不同溫度下豆漿搖瓶發酵參數值(續表5) 48
表 7、不同溫度下豆渣搖瓶發酵參數值 53
表 8、30℃藍莓發酵液添加稀釋豆漿發酵參數 68
表 9、30℃藍莓發酵液添加不同濃度豆漿發酵參數 73
表 10、30℃藍莓發酵液添加不同濃度豆漿發酵參數(續表9) 73
表 11、35℃藍莓發酵液添加稀釋豆漿發酵參數 79
表 12、35℃藍莓發酵液添加不同濃度豆漿發酵參數 84
表 13、35℃藍莓發酵液添加不同濃度豆漿發酵參數(續表12) 84

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指導教授

徐敬衡(Chin-Hang Shu)

審核日期

2021-8-30

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