臺灣博碩士論文加值系統

隨水產養殖業蓬勃發展，飼料原料中的魚粉的消耗量隨之增加。豆粕含高含量粗蛋白且供應穩定，為魚粉之最佳取代原料。但豆粕中含有植酸皂素等抗營養因子，抑制生物內養分的吸收，降低養殖水產之生長速率。本研究使用植酸酶活性乳酸菌 FPS 2520以固態發酵豆粕方式降解植酸。先探討黃豆粒徑大小與豆粕基質水分比率對植酸降解影響。黃豆粉末最有利於植酸降解，而豆粕水分比率為 1:3 有最高植酸降解率，水分含量並不影響對皂素的降解。接種 6 log CFU/g 於黃豆與豆粕中於 37oC 下發酵 72 小時後，植酸殘餘量為 20.23 ± 0.26 mg/g phytate與 54.68 ± 0.42 mg/g phytate，降解率分別為 57.92 % 與 17.06 %。利用37oC 發酵 24 小時，再升溫至 50oC 保持 24 小時之二階段溫度發酵使黃豆與豆粕之植酸降解率分別提升至 75 與 34.56 %。接著以固態發酵槽探討降解植酸之最適攪拌轉速 (0, 1, 2 rpm) 與通氣量 (0, 0.5, 1.0, 1.5, 2.0 vvm)，看能否提升 FPS 2520 發酵豆粕對植酸之降解率。轉速於 2 rpm 時使植酸降解率顯著由 51.9 提升至 61.3%，但轉速不會影響皂素降解，其皂素降解率約 65.57%。通氣量為 1.5 vvm 時不會影響菌體生長並有最低植酸殘餘量。故以 2rpm 與 1.5 vvm 為基礎探討二階段溫度發酵 50oC 時之攪拌轉速與通氣量。當於 37oC 以 2 rpm, 1.5 vvm 發酵 24 小時，再以 2 rpm, 0.5 vvm 於 50oC 發酵 24 小時之條件發酵，能使植酸降解率提升至 97.60 %，發酵後豆粕中植酸與皂素含量分別為1.16 ± 3.32 mg/g and 81.25 ± 7.53 mg/g。

The consumption of fish meal is raising because of the booming development of aquaculture. Soybean meal with stable supplement and high protein content is the best substitute for fish meal in aquaculture feed. However, soybean contains many antinutrient factors such as phytate and saponin, which can block the nutrient absorption of the organism and cause the adverse effect on fish growth. In this study Lactobacillus plantarium FPS 2520 with phytase activity was used to degrade phytate in soybean and soybean meal using a solid-state fermentation in the flask. The effects of soybean particle size and water content of substrate on the phyate and saponin degradation were evaluated first. The phytate degradation efficacy increased with decreasing size of soybean particle, with the powder form being most favored. The best ratio of soybean powder/soybean meal to water for phytate degradation is 1:3; while the water content did not affect the saponin degradation. The residual saponin content was around 1.8 to 2.0 mg/g after FPS 2520 fermentation at 37oC for 72 h, irrespective the water content of substrate and soybean particle size. After fermentation of soybean (20.23 ± 0.26 mg/g phytate) and soybean meal (54.68 ± 0.42 mg/g phytate) by FPS 2520 with an initial inoculum of 6 log CFU/mL at 37oC for 72 hr, the phytate degradation efficacy were 57.92% and 17.06 %, respectively. The phytate degradation efficacy in soybean and soybean meal was further increased to 75 and 34.56% by using two stages of incubation temperatures of 37oC for 24 h, followed by 50oC for 24 h.Using a novel solid-state fermenter to try to increase the phytate degradation efficacy in soybean meal by implication of the systems of rotation and ventilation, and also by the strategy of two incubation temperature. Factors of rotation (0, 1, 2 rpm) and ventilation (0, 0.5, 1.0, 1.5, 2.0 vvm) on phytate degradation were evaluated first. Rotation with 2 rpm significantly increased the phytate degradation efficacy from 51.9 % to 61.3 % by FPS 2520 at 37oC for 72 h; while it did not affect saponin degradation efficacy (about 65.57%). Ventilation at 1.5 vvm did not affect bacterial growth; while phytate degradation efficacy was increased from 13.56% to 38.83%. The effects of rotation and ventilation on phytate degradation were further investigated using the stratagem of 2 incubation temperatures of 37oC and 50oC. The phytate degradation efficacy in soybean meal by FPS2520 was further increased to 97.60 % using two stages of incubation temperatures of 37oC at 2 rpm and 1.5 vvm for 24 h, followed by 50oC at 2 rpm 0.5 vvm for 24 h. After fermentation the residual phytate and saponin in soybean meal was only 1.16 ± 3.32 mg/g and 81.25 ± 7.53 mg/g.

目錄
壹、前言 1
貳、文獻整理 3
一、漁業與養殖 3
二、黃豆與豆粕取代魚粉之相關研究 4
1. 黃豆與豆粕 4
2. 黃豆中的抗營養因子 4
3. 豆粕作為水產養殖飼料之研究 5
三、植酸與植酸酶 6
1. 植酸與其特性 6
2. 植酸對養殖魚類與養殖環境的影響 7
3. 植酸的去除方法 8
5. 植酸酶的來源 9
6. 植酸酶於飼料中作用降植酸之研究 11
四、皂素對養殖魚類之影響 12
五、胰蛋白酶抑制劑對養殖魚類之影響 13
六、乳酸菌 14
1. 乳酸菌對水產養殖的益處 14
2. 具植酸酶活性乳酸菌 16
3. 乳酸菌降解黃豆中的抗營養因子 16
參、實驗架構 18
肆、實驗材料與方法 19
一、實驗材料 19
1.實驗菌株 19
2.化學藥品 19
3.培養基 20
二、儀器設備 20
三、實驗方法 21
1. 菌株保存 21
2. 菌株活化 21
3.黃豆粒徑對發酵降解抗營養因子之影響 21
4.豆粕基質水分含量對發酵降解抗營養因子之影響 22
5. 37℃ 下錐形瓶發酵 22
6. 錐形瓶二階段溫度發酵 22
7. 固態發酵槽乳酸菌發酵豆粕最適降解植酸條件探討 23
8. 固態發酵槽 二階段溫度發酵豆粕最適降解植酸條件探討 23
11. 分析 24
伍、結果與討論 26
1. 錐形瓶模式探討豆粕水分含量對植酸降解之影響 26
2. 錐形瓶模式探討黃豆粒徑對植酸降解之影響 27
3. 錐形瓶模式探討豆粕與黃豆於 37。C 下發酵降解植酸與皂素 28
4. 錐形瓶模式探討豆粕與黃豆於二階段溫度發酵降解植酸與皂素 29
5.固態發酵槽模式探討轉速與通氣量對37。C下發酵降解植酸與皂素之影響 30
6.固態發酵槽探討二階段溫度發酵降解植酸與皂素之最佳轉速與通氣量 32
7. 錐形瓶及發酵槽不同轉速與通氣條件對豆粕對總多酚之影響 34
陸、結論 35
柒、參考文獻 37
捌、圖表 54
圖一、乳酸菌 FPS2520 培養於不同水份含量之豆粕 (豆粕:水) 於 37oC 發酵 72 小時之菌數 (A)、pH 值 (B)、植酸含量 (C) 及皂素含量 (D)。 54
圖二、乳酸菌 FPS 2520 發酵不同尺寸之黃豆於 37oC 下發酵 72 小時之菌數 (A)、pH值、(B) 植酸含量 (C) 及皂素含量 (D) 變化情形。 55
圖三、乳酸菌 FPS 2520 培養於黃豆與豆粕中於 37oC 發酵 72 小時之菌數 (A)、pH值 (B)、植酸含量 (C) 及皂素含量 (D)。 56
圖四、FPS 2520 固態發酵黃豆與豆粕於 37oC 下發酵 24 小時再升溫至 50oC 培養 24 小時之菌數 (A) 、pH值 (B)、 植酸含量 (C) 及皂素含量 (D) 變化情形。 57
圖五、 發酵槽轉速對乳酸菌 FPS 2520於 37oC之固態發酵槽培養豆粕 72 小時之菌數 (A)、 pH值、 (B) 植酸含量 (C) 及皂素含量 (D) 之影響。 58
圖六、 發酵槽通氣量對乳酸菌 FPS 2520於 37oC之固態發酵槽培養豆粕 72 小時之菌數 (A)、pH值 (B)、植酸含量 (C) 及皂素含量 (D) 之影響。 59
圖七、 發酵槽轉速對乳酸菌 FPS 2520於 37oC 下發酵 24 小時再升溫至 50oC 保持 24 小時培養豆粕之菌數 (A)、 pH值 (B)、植酸含量 (C) 及皂素含量 (D) 影響。 60
圖八、 通氣量速對乳酸菌 FPS 2520於 37oC 下發酵 24 小時再升溫至 50oC 保持 24 小時培養豆粕之菌數 (A)、 pH值 (B)、植酸含量 (C) 及皂素含量 (D) 影響。 61
圖九、乳酸菌 FPS 2520 培養於 (A) 豆粕與 (B) 黃豆中於 37oC 發酵 72 小時之總多酚含量。 62
圖十、 槽攪拌轉速與空氣通氣量對乳酸菌 FPS 2520 於 37oC 之固態發酵槽培養豆粕 72 小時之總多酚含量之影響。 63
圖十一、 槽攪拌轉速與通氣量速對乳酸菌 FPS 2520於 37oC 下發酵 24 小時再升溫至 50oC 保持 24 小時培養豆粕之總多酚含量影響。 64
圖十二、 槽攪拌轉速與通氣量速對乳酸菌 FPS 2520於 37oC 下發酵 24 小時再升溫至 50oC 保持 24 小時培養豆粕之總類黃酮含量影響。 65

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