冷凍是一種常用於白蝦的保鮮方式，然而不當的儲存過程中，可能會產生反覆解凍再凍結現象 (反覆凍融)，易形成大冰晶，導致白蝦蛋白質變性、質地變差以及失去保水能力等。商業上常添加冷凍保護劑延緩蛋白質冷凍變性，但商業配方中的蔗糖容易在極低溫下效果受限。因此本研究目利用海藻糖取代蔗糖，添加精油作為天然脂質抗氧化劑，並透二階反應曲面法求得 9% 海藻糖、9% 山梨糖醇與 1.6% 檸檬精油為模型中最適化冷凍保護劑濃度。並探討白蝦仁分別浸入純水 (Control)、4% 蔗糖 + 4% 山梨糖醇 (4SSo)、4% 海藻糖 + 4% 山梨糖醇 (4TSo)、9% 海藻糖 + 9% 山梨糖醇 + 1.6% 檸檬精油 (9TSoE) 和 6% 山梨糖醇 (6So) 處理，凍融 3 次後，於 4℃ 冰箱儲存下架售期與蛋白質變性之影響。微生物指標中，僅 9TSoE 組在第 7 天時的總生菌數仍低於 7 log CFU/mL；且鮮度指標中，冷藏第 11 天時，9TSoE 組的 K 值和揮發性鹽基態氮最低，分別為 30.6% 和 22.09 mg/100 g，顯示檸檬精油抑制了微生物生長，維持良好鮮度品質。而 4TSo 組儲藏至 11 天之硫巴比妥酸價顯著低於所有組別。色差變化中，儲藏第 11 天時，9TSoE 組受到檸檬精油導致蝦體色素氧化，L*、a* 和 b* 值急劇上升。在質地變化中，經冷凍保護劑處理之組別可維持硬度、彈性和咀嚼性，並在 4TSo 組觀察到更好的彈性與咀嚼性。於蛋白質微觀結構中，反覆凍融和冷藏期間蛋白質會持續變性，所有組別之表面疏水性顯著攀升，總硫醇基顯著下降。進一步透過傅里葉紅外光譜觀察於冷藏 11 天時蛋白質二級結構的變化，以 4TSo 組保有最多的 α-helix 為 15.19%，而 9TSoE 組則擁有最少的 Random coil (46.53%)。整體而言，添加精油能抑制微生物生長，然而在儲藏期間會增加蛋白質的不穩定性；4TSo 能有效保護凍融過程中的蛋白質氧化損害，且延緩儲存期間之組織蛋白結構、鮮度及品質變化。

Freezing is a common preservation method for white shrimp. However, improper storage processes may lead to freeze-thaw cycles causing issues like large ice crystal formation, protein denaturation, texture deterioration, and loss of water-holding capacity. People often added cryoprotectants to delay protein freezing denaturation. In an attempt to enhance frozen storage quality, we replaced sucrose with trehalose and added lemon essential oil as a natural lipid antioxidant. Using Response Surface Methodology (RSM), optimal concentrations were determined as 9% trehalose, 9% sorbitol, and 1.6% lemon essential oil.
Various treatment groups were compared, including immersion in pure water (Control), 4% sucrose + 4% sorbitol (4SSo), 4% trehalose + 4% sorbitol (4TSo), 9% trehalose + 9% sorbitol + 1.6% lemon essential oil (9TSoE), and 6% sorbitol (6So) to investigate the effects on shelf life and protein denaturation of white shrimp after three freeze-thaw cycles, and subsequent storage at 4℃. The 9TSoE group demonstrated the best microbial control, maintaining a total bacterial count below 7 log CFU/mL on the 7th day. It also exhibited the lowest K value and VBN on the 7th day, indicating excellent freshness preservation due to lemon essential oil's antimicrobial properties. While trehalose effectively reduced thiobarbituric acid value in the 9TSoE group, lemon essential oil in the 9TSoE group inhibited microbial growth and prevented enzymatic reactions, maintaining freshness quality. Color changes showed that 9TSoE may oxidize shrimp pigments due to the self-oxidation of lemon essential oil, leading to a sharp increase in L*, a*, and b* values on the 11th day of storage. Texture changes showed that groups treated with freezing protectants maintained hardness and cohesiveness, with the 4TSo group exhibiting superior elasticity and chewiness. Microstructural changes in proteins indicated that trehalose and sorbitol effectively protected protein denaturation and cellular structure integrity during freeze-thaw cycles, reducing freshness and quality changes during subsequent refrigerated storage. Further observation through ATR-FTIR on the 11th day of refrigeration unveiled that the 4TSo group retained the highest α-helix content at 15.19%, while the 9TSoE group exhibited the lowest random coil content (46.53%). Notably, the inclusion of essential oil proved effective in postponing lipid oxidation and inhibiting microbial growth, particularly within the initial 4 days of refrigeration. Trehalose and sorbitol (4TSo) emerged as robust protectors, preserving protein and cellular structure integrity during the freeze-thaw process and mitigating freshness and quality alterations post-freeze-thaw and subsequent refrigerated storage.

摘要 I
Abstract II
目錄 III
表目錄 VI
圖目錄 VII
壹、前言 1
貳、研究目的 2
參、文獻回顧 3
一、白蝦 3
1.1 簡介 3
1.2 銷售型態與產業現況 3
二、肌纖維蛋白質組成及其相關功能特性 3
2.1 肌纖維蛋白 3
2.2 肌動凝蛋白 (Native actomyosin, NAM) 4
三、水產品於低溫儲藏下的品質變化 4
3.1 冰晶對組織之影響 4
3.2 凍融 (Freeze-thaw, FT) 5
3.3 凍藏/凍融之肌動凝蛋白變化 5
3.4 脂質之變化 6
四、海藻糖 (Trehalose) 7
五、山梨糖醇 (Sorbitol) 7
六、檸檬精油 (Lemon essential oil) 8
七、反應曲面法 (Response surface methodology, RSM) 9
肆、實驗架構 10
一、使用反應曲面法求得最適冷凍保護劑與精油濃度條件 10
二、不同冷凍保護劑處理之白蝦仁，凍融3 次後於冷藏儲存下之品質變化 11
伍、材料方法 12
一、實驗材料 12
二、實驗藥品 12
三、實驗儀器與設備 12
四、實驗方法 13
4.1 白蝦前處理 13
4.2 冷藏儲藏 13
4.3 微生物指標 14
4.4 化學指標 15
4.5 物理指標 16
4.6 變性指標 18
4.7 統計分析 19
陸、結果與討論 20
一、以反應曲面法預測冷凍保護劑之最適條件 20
1.1 脂質氧化 20
1.2 總硫醇基含量 21
1.3 表面疏水性 21
1.4 初步最適條件評估 22
二、不同冷凍保護劑處理之白蝦，凍融 3 次後於冷藏儲存下之品質變化 22
2.1 微生物指標 22
2.2 化學指標 23
2.3 物理指標 26
2.4 變性指標 30
柒、結論 35
捌、參考文獻 36
玖、圖表 45

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