臺灣博碩士論文加值系統

本研究將進行納豆菌 (Bacillus subtilis) 在紅藜的發酵試驗，以曲面反應法評估產生納豆激酶之最佳發酵條件，結果顯示接菌量6 log CFU/g、發酵時間36小時、發酵溫度37 ℃為最佳發酵組合條件。進一步分析發酵對紅藜之酚類、類黃酮、甜菜苷等成分、抗氧化能力、色澤、以及理化特性的影響，並以純水及60%乙醇進行紅藜及發酵紅藜的成分萃取，發現Bacillus subtilis發酵可顯著提升紅藜的抗氧化成分與抗氧化能力，未發酵紅藜之總酚、類黃酮為281 GAE/100 mg與229 QuE/100 mg，發酵後分別提升為554 GAE /100mg與481 QuE/100mg。透過HPLC分析顯示Feruloc acid、Rutin、Catechin是紅藜中含量最高的酚酸，在未發酵紅藜的濃度分別是153.90 mg/ml、135.35 mg/ml、84.67 mg/ml，發酵後提升為175.80 mg/ml、164.80 mg/ml、105.94 mg/ml，但兩種主要的甜菜色素Betanin、IsoBetanin，則是在發酵後濃度顯著的降低。而總抗氧化能力、還原力、DPPH抑制能力的分析，也發現類似的趨勢，發酵使紅藜的純水萃取物及60%乙醇萃取物，三種抗氧化能力都有顯著性的提升。在物性方面，未發酵紅藜之硬度為2509 g，發酵後降低為1439 g；而發酵前的黏度為40.18 g，發酵後提升為140.80 g。以上結果顯示，透過B. subtilis發酵開發紅藜納豆，可提升抗氧化成分之濃度，並賦予納豆機酶活性，且發酵可調整其軟硬度之適口性，具有開發為機能性高齡食品之應用潛力。

In this study, the fermentation test of Bacillus subtilis in red quinoa will be conducted, and the optimal fermentation conditions for the production of nattokinase will be evaluated by response surface methodology. The results show that the inoculation amount of 6 log CFU/g, the fermentation time of 36 hours, and the fermentation temperature of 37 oC is the best fermentation combination condition for production nattokinase. Further analyze the effects of fermentation on the phenols, flavonoids, betain and other components, antioxidant capacity, color, and physical properties in red quinoa, and extract the components of red quinoa and fermented red quinoa with water and 60% ethanol. Fermentation of B. subtilis can significantly enhance the antioxidant components and antioxidant capacity of red quinoa. The total phenols and flavonoids of unfermented red quinoa are 281 GAE/100mg and 229 QuE/100 mg, and after fermentation, the total phenols and flavonoids are increased to 554 GAE/100mg and 481 QuE/100 mg, respectively. HPLC analysis showed that feruloc acid, rutin, and catechin are the major phenolic acids in red quinoa, and the concentrations of unfermented red quinoa are 153.90 mg/ml, 135.35 mg/ml, 84.67 mg/ml, and after fermentation, the concentration of them are increased to 175.80 mg, 164.80 mg/ml, and 105.94 mg/ml, respectively. However, the concentration of the two main beet pigments Betanin and IsoBetanin decreased significantly after fermentation. The analysis of total antioxidant capacity, reducing power, and DPPH inhibition capacity also found a similar trend. Three antioxidant capacities have been significantly improved by B. subtilis fermentation in red quinoa. In terms of physical properties, the hardness of unfermented red quinoa is 2509 g, which is reduced to 1439 g after fermentation; the viscosity before fermentation is 40.18 g, and the viscosity is increased to 140.80 g after fermentation. The above results show that the development of red quinoa natto through B. subtilis fermentation can increase the concentration of antioxidant components, and endow natto with nattokinase activity, and the fermentation can adjust its softness and palatability, and it can be developed as a functional elderly food.

摘要....i
Abstract....ii
誌謝....iv
目錄....v
表目錄....vii
圖目錄....viii
第一章 文獻探討....1
一、紅藜的背景....1
二、紅藜的利用及開發....3
三、紅藜中的酚類化合物....9
四、納豆簡介....13
五、枯草桿菌....13
六、納豆激酶....16
七、納豆發酵製品的健康益處....18
八、反應曲面法....20
九、研究目的....21
第二章 材料與方法....23
一、實驗原料與菌株....23
二、紅藜納豆樣品測試納豆激酶含量....23
三、反應曲面法....23
四、血纖維蛋白活性測定....24
(一) 納豆激酶酵素活性測定....24
(二) 納豆激酶酵素活性計算....24
五、血纖維蛋白平板試驗....25
(一) 試藥配置....25
(二) 實驗流程....25
六、納豆激酶納豆菌發酵萃取液的人工血栓溶解活性....25
七、紅藜納豆樣品抗氧化成分製備....26
八、萃取....26
九、總酚含量測定....26
十、類黃酮含量測定....26
十一、甜菜色素含量測定....27
十二 酚酸及甜菜苷含量測定....27
十三、總抗氧化能力....31
十四、還原力....31
十五、DPPH自由基清除能力試驗....32
十六、水分含量及水活性測定....33
十七、色澤分析....33
十八、物性結構儀分析及黏度測定....33
十九、統計分析....35
第三章 結果....36
一、納豆菌生長曲線....36
二、納豆激酶活性....36
三、RSM條件分析....40
四、血纖維平板試驗....43
五、總酚含量....44
六、類黃酮含量....44
七、甜菜色素含量....44
八、酚類化合物分析....48
九、甜菜色素分析....53
十、還原力能力分析....57
十一、DPPH能力試驗分析....58
十二、總抗氧化能力分析....61
十三、水分及水活性測定....63
十四、色澤測定....63
十五、物性分析....63
第四章 討論....67
第五章 結論....70
參考文獻 ....72
Extended Abstract....86
Abstract....86

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