臺灣博碩士論文加值系統

本實驗以全豆利用之概念，使用酵素水解綠豆，併用 Lactobacillus johnsonii BCRC 17010 或 Lactobacillus plantarum subsp. BCRC 10069 發酵，探討酵素水解與併用益生菌發酵對綠豆營養價值提升與保健機能性。結果顯示 1% 纖維素酶與 1% 澱粉酶同時添加於綠豆泥，以 50oC 水解 2 小時，經 Lactobacillus johnsonii BCRC 17010 或 Lactobacillus plantarum subsp. BCRC 10069 發酵 24 小時，菌數達 9.36 及 8.59 log CFU/mL；pH 值下降至 3.77 與 4.11。經掃描式電子顯微鏡觀察，綠豆水解物或發酵物之纖維結構明顯被破壞。總酚、類黃酮及花青素含量經水解後顯著增加；總胺基酸含量較未水解組增加 16% (52.8~61.24 g/100 mL)，經 Lactobacillus plantarum subsp. BCRC 10069 發酵 24 小時後，總胺基酸含量增加 7% (65.56 mg/100 mL)，顯示綠豆的生物活性成分可經由水解或發酵過程釋放出來。綠豆水解物與發酵物之清除 ABTS+• 自由基、抑制 α-澱粉酶及 α-葡萄糖苷酶活性之 IC50 值皆有顯著下降，分別為 0.47~1.52 mg/mL、1.89~2.14 mg/mL、35.85~45.89 mg/mL。經 GC/MS 分析之主要機能性成份為甲基麥芽酚 (Maltol) 與鄰苯二酚 (Catechol)，其清除 ABTS+• 自由基之 IC50 值分別為 1.41 與 2.52 μg/mL。綠豆水解發酵物進行耐酸性與耐膽鹽試驗，於 pH 3.0 PBS 耐酸 3 小時之菌數分別維持在 9.13~9.15、8.13~8.20 log CFU/mL，膽鹽耐受性達 97.64%、95.37%。二種發酵物之安姆氏試驗結果顯示，無論添加活化代謝物質與否，對測試菌株 Salmonella typhimurium TA97a、TA98、TA100、TA102 及 TA1535 皆不會產生基因毒性。利用脂多醣 (LPS) 刺激小鼠巨噬細胞 (RAW 264.7) 產生發炎反應，在添加 40 mg/mL 之綠豆未水解物、水解物與二種發酵物處理 24 小時條件下，可抑制 LPS 誘發之一氧化氮 (36.80~90.40%)、介白素-1β (49.26~72.29%) 及介白素-6 (17.78~93.75%) 生成。由上述結果顯示，綠豆利用酵素水解或併用益生菌發酵有助於機能性成分釋出並提升其保健機能性。

This study aimed to improve the functionality of mung bean, sample was hydrolyzed by cellulase and amylase, and further fermented by Lactobacillus johnsonii BCRC 17010 or Lactobacillus plantarum subsp. BCRC 10069. After 2 hr hydrolysis at 50oC by the combination of 1% cellulase and 1% α-amylase and further 24 hr fermentation with Lactobacillus johnsonii BCRC 17010 or Lactobacillus plantarum subsp. BCRC 10069 at 37oC, the LAB counts increased to 9.36 and 8.59 log CFU/mL, and the pH declined to 3.77 and 4.11, respectively. Scanning electron microscope (SEM) photograph indicated that the obvious break occurred in fibers after hydrolysis and fermentation. Increases in the amounts of the total phenolic, flavonoid and anthocyanin content and total amino acid content of hydrolysates and fermented samples were observed, compared with those of non-hydrolysed samples. These phenomena suggested the release of bionutrients occurred after hydrolysis or fermentation process. In hydrolyzed and further fermented samples, significant decreases in half maximal inhibitory concentration (IC50) on ABTS+˙ radical scavenging (0.47~1.52 mg/mL), α-amylase (1.89~2.14 mg/mL) and α-glucosidase (35.85~45.89 mg/mL) were observed. Gas chromatography-mass spectrometry analysis revealed that the major components were maltol and catechol. The IC50 on ABTS+• radical scavenging were 1.44 and 2.52 μg/mL, respectively. The fermented mung bean hydrolysates maintained 9.13~9.15 log CFU/mL (97.64%), 8.13~8.20 log CFU/mL (95.37%) in acid resistance (PBS at pH 3.0) and bile tolerance tests, respectively. According to the Ames tests, obtained, no matter whether the activating metabolites were added or not, the fermented products did not cause the genomic toxicity on the tested microbes ( Salmonella typhimurium TA97a, TA98, TA100, TA102 and TA1535). The inhibition of LPS-induced in murine macrophage cell (RAW 264.7) were used to investigate the anti-inflammatory activities of the hydrolysates and their fermented samples. Inhibition in the releasing of NO (36.80~90.40%), interleukin 1β (49.26~72.29%) and interleukin 6 (17.78~93.75%) were obtained, respectively. These data suggested that hydrolysis and fermentation have high potential to promote the releases of functional components, which could subsequently improve the functionality of mung bean.

謝誌
摘要
Abstract
目錄
表目錄
圖目錄
壹、研究目的與背景
貳、文獻回顧
一、綠豆 (Vigna radiata L.)
(一)綠豆之簡介與品種特性
(二)綠豆之營養與生理功能
二、酵素種類 (Enzyme type)
(一)纖維素酶 (Cellulase)
(二)澱粉酶 (Amylase)
三、益生菌 (Probiotic)
(一)益生菌之特性
(二)益生菌之種類
(三)益生菌之益處與應用
四、醣解酵素
(一)α-澱粉酶 (α-amylase)
(二)α-葡萄糖苷酶 (α-glucosidase)
五、自由基與活性氧化物 (Free radical and reactive oxygen species )
(一)自由基與活性氧化物對生理功能的影響
(二)抗氧化劑作用機轉
(三)天然抗氧化劑 (Natural antioxidant)
六、發炎反應 (Inflammatory response)
(一)巨噬細胞 (macrophage) 與發炎反應相關性
(二)發炎反應之介質 (mediators)
參、實驗設計
肆、實驗材料與方法
一、實驗材料
(一)原料與菌株
(二)主要藥品
(三)主要儀器
二、實驗方法
(一)水解條件探討
(二)益生菌發酵綠豆水解物試驗
(三)一般成份分析
(四)醣類成份分析
(五)機能性成份分析
(六)抗氧化能力
(七)蛋白質組成分析
(八)降血糖能力試驗
(九)綠豆發酵物之機能性分析
(十)綠豆水解物與發酵物之體外抗發炎能力分析
(十一)統計分析
伍、結果與討論
一、綠豆最適水解條件
二、纖維素酶與澱粉酶水解綠豆之結構變化
三、綠豆未水解或酵素水解併用益生菌發酵期間之結構變化
四、綠豆未水解或酵素水解併用益生菌發酵期間之 pH 值與菌數變化
五、綠豆水解物或酵素水解併用益生菌發酵期間之成份變化
六、益生菌發酵綠豆水解物之耐酸與耐膽鹽能力
七、綠豆水解物或酵素水解併用益生菌發酵期間之抗氧化能力
八、綠豆水解物或酵素水解併用益生菌發酵期間之抑制 α-amylase 與 α-glucosidase 活性能力
九、機能性成分定性與定量分析
十、安姆氏試驗 (Amest test)
十一、綠豆水解物與發酵物之體外抗發炎能力分析
陸、結論
柒、參考文獻

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