臺灣博碩士論文加值系統

轉麩胺酸基胺基酶 (Transglutaminase; TGase) 能促使蛋白質間產生 -(-glutamyl) lysine 鍵結而形成分子內或分子間的共價結合，故本研究利用此酵素作為盒裝豆腐 (packed tofu) 之凝膠劑 (coagulant)，探討不同來源 TGase 對其品質之影響。保水率 (water holding capacity) 以市售豆腐的77.5% 為最高，其次為添加0.5% TG-B (46-76 U/g)、0.5% TG-K (78-126 U/g) 及2.45 % MTGase (817.2 U/g) 的 67.6 %、66.4 % 及57.9%。以 1% MgCl2 混合 0.5% TG-K 為凝膠劑，經物性儀測得較好膠強度 (gel strength)，為 1328 gxmm，高於市售組的866.0 gxmm；0.3% TG-B 與 0.3% MgCl2 組之膠強度為 750.0 gxmm，使用2.45 % MTGase (817.2 U/g) 時膠強度為 150.1 gxmm。混合添加 MTGase，可提高僅添加 MgCl2 時的膠強度，由 218.7提升至 334.0 gxmm。經電子顯微鏡掃描圖 (scanning electron microscope; SEM) 發現市售豆腐均勻度佳且孔隙小，MTGase 組則較為平整且出現絲狀般結構連結膠體。若使用 MTGase 可改善 MgCl2 及 CaSO4 均勻度並解決孔隙大的問題。為確立 MTGase之鍵結，利用 LC-MS (Liquid chromatography–mass spectrometry) 圖譜分析，可增加 45% -(-glutamyl) lysine 鍵，證實形成 G-L-bounds。此外為了解自製豆腐被消化酵素分解情形，經體外消化性試驗 (in vitro digestion) 處理後，MTGase 組其可溶性蛋白質 (soluble protein)、胜肽 (peptide) 及游離氨基酸 (free amino acid) 量分別增加 277.2、12.6 及 1.46 (mg/g tofu)，與市售豆腐的 261.7、17.7 及 3.92 (mg/g tofu) 並無顯著差異 (p&;lt;0.05)；進一步分析各游離胺基酸，MTGae 組之glutamic acid、lysine 及 glutamine 分別增加 9.45、0.29 及 0.18 (mg/100g)，市售組則分別為 2.44、1.05 及 1.14 (mg/100g)(p&;lt;0.05)，顯示 -(-glutamyl) lysine 此鍵結可被胃蛋白酶 (pepsin) 及胰蛋白酶 (trypsin) 所水解。

This study aimed to investigate the effect of microbial TGase (MTGase) on the　texture of packed tofu and the possibility of using MTGase as coagulant. The highest water holding capacity was 77.5% from commercial tofu, while that of with 0.5% TG-B (46-76 U/g), 0.5% TG-K (78-126 U/g), and 2.45 % MTGase (817.2 U/g) were 67.6 %, 66.4 % and 57.9%, respectively. Combination use of 0.5% commercial TGase (TG-K) with 0.1% MgCl2 had gel strength 1328 gxmm which were higher than 866.0 gxmm of commercial tofu. The gel strength of sample with 0.3 % commercial TGase (TG-B) and 0.3% MgCl2 was 750.0 gxmm, while that with 2.45 % MTGase (817.2 U/g) was 150.1 gxmm. Comparing with that was using MgCl2 per se, the gel strength of samples with MTGase increased from 218.7 to 334.0 gxmm. Comparing the consistency of texture and porosity in scanning electron microscope (SEM) diagrams, the commercial tofu was the best, then the samples with MTGase > CaSO4 > MgCl2. SEM profiles suggested that MTGase could improve the coarse texture of tofu. According to LC-MS profile, 45% of -(-glutamyl) lysine bonds of samples using MTGas as coagulant increase, comparing to those without MTGase. After being digested by trypsin and pepsin, no significant differences in soluble protein, peptides and free amino acids was observed between samples with MTGase (277.2, 12.6 and 1.46 mg/g, respectively) and commercial tofu (261.7, 17.7 and 3.92 mg/g, respectively) (p&;lt;0.05). No significant differences of increasing values in glutamic acid, lysine and glutamine between samples with MTGase (9.45, 0.29 and 0.18 mg/100g, respectively) and commercial tofu (2.44, 1.05 and 1.14 mg/100g, respectively) was also obtained (p>0.05), suggesting the -(-glutamyl) lysine isopeptide bonds can be digested by trypsin and pepsin.

中文摘要 1
Abstract 2
壹、研究背景與目的 3
貳、文獻整理 4
一、轉麩胺酸醯胺基酶 4
(一)、TGase的分布 4
(二)、TGase 的分子特性 5
(三)、TGase的催化作用 5
(四)、微生物產生的MTGase 6
(五)、TGase的活性測定 6
(六)、影響TGase活性作用因子 7
(七)、TGase於蛋白食品工業之應用 9
(八)、MTGase之安全性 9
二、大豆之簡介 10
(一)大豆來源 10
(二)大豆的成分組成 10
(三)、大豆加工製品 11
(四)、豆腐 12
(五)、豆腐凝膠機制 12
(六)、豆腐的製作及種類 13
(七)、影響豆腐產率及品質之因素 13
(八)、豆腐加工過程與異黃酮素含量變化 15
(九)、豆腐的品質評估指標 15
(十)、TGase 應用於豆腐製備 16
三、質地分析 (texture profile analysis) 16
叁、研究架構 18
肆、實驗材料與方法 19
一、酵素來源 19
二、原料 19
三、化學藥品 19
四、儀器 20
五、實驗方法 21
(一)、盒裝豆腐 (Packed tofu) 製備 21
(二)、酵素製備 22
(三)、酵素活性測定 22
(四)、凝膠劑 22
(五)、大豆浸泡時間 23
(五)、pH值分析 23
(六)、保水力分析 (Water holding capacity) 23
(七)、質地分析 (texture profile analysis) 23
(八)、ε-(γ-glutamyl) lysine 測定 24
(九)、掃描式電子顯微鏡之觀察 (Scanning Electron Microscope analysis) 24
(十)、體外消化性試驗 (In vitro digestion) 24
(十一)、可溶性蛋白質測定 24
(十二)、胜肽測定 25
(十三)、總游離胺基酸測定 25
(十四)、游離胺基酸測定 25
(十五)、總生菌數測定 26
(十六)、統計分析 26
伍、結果與討論 27
一、大豆浸泡吸水量 27
二、以轉麩胺酸醯胺基酶做為凝膠劑之條件建立 27
三、利用不同來源TGase做為凝膠劑 27
四、不同凝膠劑對豆腐細微結構之影響 30
六、ε-(γ-glutamyl) lysine 分析 33
七、MTGase對於豆腐儲存性之影響 33
陸、結論 34
柒、參考文獻 35

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