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研究生(外文):Chiung-Wen Yeh
論文名稱:應用義大利白松露Tuber magnatum生產多醣體最適化培養之探討
論文名稱(外文):Optimization of the cultivation conditions on the Polysaccharide production from Tuber magnatum
外文關鍵詞:Tuber magnatumsubmerged culturepolysaccharideResponse surface methodology
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近年來,應用液態醱酵培養菇類生產菌體和多醣體,已成為菇類利用之新趨勢。本研究的目的是以義大利白松露菌Tuber magnatum 開發液態醱酵生產其菌絲體與胞外多醣體,在研究結果中得知,碳、氮源對菌絲體生長與其代謝物生產之影響極大。在不同碳源測試實驗中,以sucrose為最佳碳源,其生產胞外多醣體為最高 397.6 mg/L,在不同氮源測試中,以yeast extract為最佳氮源,其生產胞外多醣體為最高 706.55 mg/L。
在基質最佳化實驗中,選擇碳源、氮源、溫度、pH、鎂離子及鉀離子為設計因子以尋找最佳化之培養基,利用回應曲面法來找出最佳的培養基組成,經由回應曲面法實驗設計法,結果顯示以sucrose 10.60 g/L、yeast extract 10.92 g/L、MgSO4 1.0 g/L、KH2PO4 1.01 g/L,培養基最適初始pH和醱酵溫度,分別為6.5和25.0℃在恆溫震盪箱以100 rpm的操作條件下,可得到最佳的胞外多醣體 1329.25 mg/L,胞外多醣體為未最佳化前的3.34倍。
Recently, the submerged fermentation process for the production of mushrooms’ mycelia and polysaccharides has been developed and become the new trends for the usage of mushrooms. Truffle with its special aroma and taste in food market, has a higher economic value that all the other mushrooms. In this study, the main goal is to enhance the growth of Tuber magnatum to produce Tuber mycelia and polysaccharides in submerged culture. In the results, the carbon and nitrogen sources in the medium had significant influences on the mycelium and metabolites. In carbon sources tests, sucrose had the best Tuber polysaccharides production, which reached at 397.6 mg/L. In nitrogen sources tests, yeast extract was identified to be the most favorable source for Tuber polysaccharides production, which reached at 706.5 mg/L.
In the optimization of fermentation medium, carbon source, nitrogen source, temperature, pH, magnesium ion and potassium ion were chosen as the factors of designing the best formula for growth medium. According to the response surface methodology, the optimal concentrations of sucrose, yeast extract, magnesium sulfate and potassium dihydrogen phosphate was 10.6, 10.92, 1.0 and 1.0 g/L, respectively. The fermentation condition were conducted at 25℃ and pH 6.5 on a rotary shaker at 100 rpm. The validation experiment showed the experimental values were in good agreement with the predicted values. The highest EPS production (1329.25 mg/L) was obtained under the optimal conditions, which was 3.3 fold higher than that of the control.
中文摘要 I
Abstract II
表目次 VI
圖目次 VII
一、緒論 1
(一) 前言 1
(二) 研究動機 2
二、文獻回顧 3
(一) 松露菌(Truffle) 3
1. 松露之簡介 3
2. 松露生長環境及栽培方法 6
3. 松露於液態培養中的球狀菌絲體的形成 7
(二) 松露多醣體 9
1. 多醣體之簡介 9
2. 多醣體之療效 12
3. 抗腫瘤機制 13
4. 多醣體之結構 15
5. (1,3)-β-D-葡聚醣 16
(三) 反應曲面法 (response surface methodology, RSM) 19
1. 極值點區域逼近 20
2. 反應曲面數學模式建立 20
3. 數學模式適切性檢驗 21
4. 極值點之決定 21
5. 各因子影響效應分析 22
三、材料與方法 23
(一) 實驗設備 23
1. 實驗儀器 23
2. 實驗藥品 24
(二) 菌種 25
1. 實驗菌株 25
2. 菌株培養 25
(三) 培養基組成 25
1. 前培養培養基 25
2. 主培養培養基 26
(四) 搖瓶培養之試驗 27
1. 不同碳源之培養 27
2. 不同碳源濃度的培養 27
3. 不同氮源之培養 27
4. 不同氮源濃度的培養 28
5. 不同起始pH值的培養 28
(五) 實驗分析 31
1. 乾重分析法: 32
2. 葡萄糖分析方法 32
3. 殘留糖濃度及純度的測定 33
4. 分子量的測定 34
5. 水溶性多醣濃度測定 34
四、結果與討論 36
(一) 碳源之搖瓶試驗 36
(二) Sucrose濃度之影響 40
(三) 氮源之影響 44
(四) Yeast extract濃度之影響 48
(五) 起始pH值之影響 52
(六) 回應曲面法於多醣生產最適培養基之探討 56
五、結論與展望 65
(一) 結論 65
(二) 未來與展望 66
參考文獻 67
附錄 72
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