臺灣博碩士論文加值系統

醬油是一種普遍且具亞洲特色的調味品，傳統發酵釀造醬油製造過程中所產生的副產物─醬油粕年產量約 1.3 萬噸，主要成分為大豆和小麥，蛋白質含量高，適合作為開發活性胜肽基材之原料。蛋白質透過酵素水解所獲得之胜肽具有DPP-IV 抑制、ACE 抑制及抗氧化能力等功效，因此本研究目的為醬油粕蛋白質鑑定及其酵素水解物之潛在生理活性探討。經由鹼萃取純化之醬油粕分離蛋白透過十二烷基硫酸鈉聚丙烯醯胺凝膠電泳、質譜儀等蛋白質體學方式鑑定其蛋白質，經 Mascot 資料庫分析後依據其序列覆蓋率，挑選鹼性 7S 球蛋白、大豆球蛋白G1和 β-伴大豆球蛋白 β-亞基 1，接著以 BIOPEP-UWM 資料庫進行潛在活性胜肽分析和模擬酵素水解。醬油粕分離蛋白以木瓜酵素、胃蛋白酶及鳳梨酵素最適條件下進行水解，並測試其抗氧化及 ACE 抑制能力。木瓜酵素及鳳梨酵素水解物有較高金屬螯合能力及還原力，而胃蛋白酶水解物則具有最高 DPPH 自由基清除率及 ACE 抑制活性 72.26%，並透過超過濾區分其分子量 (<1 kDa、1-5 kDa、5> kDa)，以胃蛋白酶水解物 <1 kDa ACE 抑制活性 63.19% 為最高，顯示分子量愈小的胜肽愈具有 ACE 抑制能力。綜合上述結果，醬油粕具有發展降血壓及抗氧化胜肽的潛力，未來可以進一步分析純化並定序降血壓及抗氧化胜肽。

Soy sauce is characteristic and universal condiment in Asia, the annual yield of by-products from traditional soy sauce industries are nearly thirteen thousand tons in Taiwan. The fermented by-products from soybean and wheat contain abundant protein. The proteins hydrolyzed by the enzyme can obtain the bioactive peptides such as DPP-IV inhibitor, ACE inhibitor and antioxidant peptides. This study aimed to analyze the anti-hypertensive and antioxidative potential of bioactive peptides from fermented by-products of soybean and wheat. Alkali fractions of soy sauce by-products were identified by proteomic techniques, followed by SDS-PAGE and mass spectrometry with Mascot database. Based on the sequence coverage and molecular weight, protein basic 7S globulin, glycinin G1 and beta-conglycinin beta subunit 1 were chosen to analyze bioactive peptides with BIOPEP-UWM database. The results showed fermented by-products of soybean hydrolyzed by papain, pepsin and bromelain obtained higher ACE inhibition and antioxidant peptides. The pepsin hydrolysate had the highest ACE inhibitory activity 72.26%. The molecular weight of pepsin hydrolysates was differentiated (<1 kDa、1-5 kDa、5> kDa), and the result showed that peptides with MW <1 kDa had the best ACE inhibitory activity 63.19%. Overall, the pepsin hydrolysate had potential to produce antioxidant and ACE inhibitory peptides, and can be purified to identify the sequences of the amino acids in the future study.

壹、前言 P1
貳、文獻回顧 P2
一、醬油 P2
1.市售調味品之比較 P2
2.醬油副產物—醬油粕 P4
二、植物性蛋白 P4
1.大豆 P4
2.小麥 P5
三、蛋白質分離方法 P6
1.等電點沉澱 P6
2.十二烷基硫酸鈉聚丙烯醯胺凝膠電泳 (SDS-PAGE) P6
3.超過濾 P6
四、蛋白質鑑定 P7
1.質譜儀鑑定技術 P7
2.質譜儀鑑定技術之應用 P8
五、蛋白質酵素水解 P8
1.資訊工具分析 P9
2.苦味胜肽 P9
3.活性胜肽 P10
六、生理活性 P12
1.抗氧化 P12
2.降血壓 P13
3.降血糖 P13
4.神經系統調節 P14
5.免疫系統調節 P14
參、實驗架構與材料方法 P15
一、實驗材料 P17
二、實驗方法 P17
1.成分分析 P17
2.樣品前處理 P18
3.鹼萃取 P18
4.十二烷基硫酸鈉聚丙烯醯胺凝膠電泳 (SDS-PAGE) P18
5.蛋白質鑑定 P19
6.生物資訊工具分析 P21
7.酵素水解 P22
8.胜肽生理活性測試 P23
8.1抗氧化能力 P23
8.2降血壓活性 P24
9.超過濾 P25
10.統計分析 P25
肆、結果與討論 P26
一、醬油粕成分 P26
二、醬油粕蛋白質鑑定 P26
三、醬油粕蛋白質模擬酵素水解 P28
1.潛在活性胜肽 P28
2.模擬酵素水解 P28
四、醬油粕酵素水解物及其生理活性探討 P29
1.水解物特徵 P29
2.抗氧化能力 P30
3.ACE 抑制能力 P33
伍、結論 P35
陸、未來研究方向 P36
柒、參考文獻 P37
捌、圖表 P44

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