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研究生:林靖超
研究生(外文):Ching Chao Lin
論文名稱:添加脂肪酸對液態培養巴西蘑菇菌絲體的抗氧化能力及多醣體之影響
論文名稱(外文):Effect of Fatty Acids Addition on the Antioxidant Activity and Polysaccharides by the Mycelium of Agaricus blazei Murill in Submerged Liquid Culture
指導教授:李 柏 旻謝 欽 城
指導教授(外文):Pomin LiChing-Chen Hsieh
學位類別:碩士
校院名稱:國立屏東科技大學
系所名稱:生物機電工程系
學門:工程學門
學類:機械工程學類
論文種類:學術論文
論文出版年:2006
畢業學年度:94
語文別:中文
論文頁數:75
中文關鍵詞:巴西蘑菇脂肪酸菌絲覆蓋率胞外多醣抗氧化能力
外文關鍵詞:Agaricus blazei Murillfatty acidThe mycelia coverageexopolysaccharideantioxidant activity
相關次數:
  • 被引用被引用:3
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巴西蘑菇為食藥用真菌,於液態深層培養所產生之二次代謝物具有抗腫瘤活性、提高人體免疫力及抗氧化能力等生理活性。本研究將討論脂肪酸添加於培養基對巴西蘑菇菌絲體的菌絲生長、胞外多醣含量及菌絲體抗氧化能力等影響。結果顯示,以不同種類及濃度之脂肪酸添加於YMA固態培養基,對菌絲的生長速度差異性不大,但添加Linoleic acid的菌絲生長型態與其餘鬆散生長方式不同,所表現的型態為內部及邊緣緊實的完整型態。於搖瓶培養巴西蘑菇菌絲的試驗中,添加Stearic acid、Palmitic acid及Oleic acid於培養基對菌絲生長顯現了促進效果,添加Linoleic acid於低濃度時(0.5 mg/ml)便顯現了抑制現象,高於1.0 mg/ml之後幾乎無新增生之菌絲球形成。隨著培養基中Stearic acid及Palmitic acid的濃度增加,醱酵後的液態培養基之pH值會隨之降低,胞外多醣含量也隨之增加。然而,培養基的pH值隨Linoleic acid添加濃度增加,菌絲及胞外多醣量則隨脂肪酸添加濃度而降低。Oleic acid的最高菌絲濃度及胞外多醣量發生在濃度2.0 mg/ml時。
在抗氧化力試驗當中,添加少量的Oleic acid及Palmitic acid(0.5 mg/ml及1.0 mg/ml)所培養的巴西蘑菇菌絲有較佳的抗氧化力。在還原力試驗當中,添加與未添加脂肪酸的的表現無顯著差異。而各種脂肪酸的菌絲之間,用乙醇萃取濃度2.5 mg/ml下,添加1.0 mg/ml亞油酸的巴西蘑菇菌絲體之還原力吸光值最高。其次為添加0.5 mg/ml Palmitic acid的巴西蘑菇菌絲體。添加Linoleic acid的菌絲與未添加脂肪酸的菌絲在高乙醇萃取濃度下有較佳的捕捉DPPH自由基能力。乙醇萃取濃度在2.5 mg/ml~10 mg/ml時,添加Linoleic acid的巴西蘑菇菌絲表現較高的螯合亞鐵離子能力,其次是未添加脂肪酸的菌絲。掃除超氧陰離子能力部分,以未添加脂肪酸的巴西蘑菇菌絲體較好。
添加脂肪酸的類型及濃度對於巴西蘑菇菌絲體的菌絲乾重及多醣含量有促進或抑制現象,菌絲乾重及多醣含量呈現正相關性,但不與抗氧化性質呈現相對應關係。
Agaricus blazei Murill is an edible and can be used as a medicinal fungus. The secondary metabolites produced from the submerged culture of mycelium were identified as antitumor, immunity-increasing and antioxidant activity. The growth of mycelium,content of exopolysaccharide and antioxidant activity of Agaricus blazei Murill mycelium with different fatty acids in liquid medium were discussed. Results of the study showed that without notable difference in the growth rate of mycelium with different fatty acids and different concentration in solid medium. However, The internal and rim of mycelium were closely knit with Linoleic acid in solid medium and different with other types. Stearic acid, Palmitic acid and Oleic acid addition showed a stimulatory effect in mycelial growth, but showed a inhibition effect with Linoleic acid at low concentration(0.5 mg/ml) and the mycelium without proliferation when Linoleic acid addition above 1.0 mg/ml. As the concentration of Stearic acid and Palmitic acid increases, the mycelium and content of exopolysaccharide increases with the pH value of broth decreases in flask cultured. As the concentration of Linoleic acid increases, the mycelium and content of exopolysaccharide decreases with the pH value of broth increases in flask cultured. The higher concentration of mycelium and exopolysaccharide with Oleic acid in liquid medium at 2.0 mg/ml.

The mycelium of Agaricus blazei Murill cultured with Oleic acid and Palmitic acid (0.5 mg/ml~1.0 mg/ml) show a good antioxidant activity (AOA). That is not notable difference with fatty acid addition or not in reduce power. The highest absorbance of reduce power from the mycelium of Agaricus blazei Murill with Linoleic acid at 1.0 mg/ml and EtOH extract at 2.5 mg/ ml. The secondary were Palmitic acid at 0.5 mg/ml. The mycelium of Agaricus blazei Murill cultured with Linoleic acid and without fatty acids addition at high EtOH extract show a good scavenging effect on 1,1-diphenyl-2-picrylhydrazyl radicals. The mycelium of Agaricus blazei Murill cultured with Linoleic acid at EtOH extract (2.5 mg/ml~10 mg/ml) showed a high chelating effects on ferrous ions, The secondary were the mycelium cultured without fatty acids. The mycelium cultured without fatty acids showed a good superoxide anion scavenging activity.
The extent of stimulation or inhibition of the mycelial growth and polysaccharide formation by Agaricus blazei Murill were associated with types and concentration of fatty acids. Mycelia dry weight and exopolysaccharide of Agaricus blazei Murill has straight dependence but without relatedness in antioxidant properties.
摘 要 I
ABSTRACT III
誌 謝 V
目錄 VII
表目錄 IX
圖目錄 XI
第1章 前言 1
第2章 文獻探討 2
2.1 食藥用菇類簡介 2
2.2 食藥用菇類的生理活性 3
2.3 巴西蘑菇的介紹 4
2.3.1 巴西蘑菇的分類地位及型態特徵 4
2.3.2 巴西蘑菇之化學成分 4
2.3.3 巴西蘑菇之多醣體 5
2.3.4 巴西蘑菇之抗氧化及活性物質 7
2.4食藥用菇類液態培養條件 7
2.4.1 培養基組成 8
2.4.1.1 碳源 8
2.4.1.2 氮源 8
2.4.1.3 碳氮比 9
2.4.1.4 無機鹽類 10
2.4.2 攪拌速率 10
2.4.3 溫度 10
2.4.4 pH值 10
2.4.5黏度(viscosity) 11
2.4.6其他添加物 11
2.4.7溶氧值(DO) 12
2.5脂肪酸的介紹 12
2.5.1脂肪酸的類型及性質 12
2.5.2脂肪酸及植物油的添加對食藥用菇類之影響 14
第3章 材料與方法 15
3.1實驗材料 15
3.1.1巴西蘑菇菌株 15
3.1.2固態培養基(YMA) 15
3.1.3液態醱酵培養基(YM) 15
3.1.4實驗藥品 15
3.1.5實驗儀器 17
3.2實驗流程架構 18
3.3實驗方法 19
3.3.1巴西蘑菇菌絲體培養 19
3.3.1.1巴西蘑菇菌絲體保存 19
3.3.1.2巴西蘑菇菌絲體活化 19
3.3.2巴西蘑菇菌絲體固態培養試驗 19
3.3.2.1菌絲體生長試驗 19
3.3.2.2不同脂肪酸對菌絲體生長之影響試驗 19
3.3.3巴西蘑菇液態醱酵培養 20
3.3.3.1搖瓶震盪培養實驗 20
3.3.3.2醱酵槽培養實驗 20
3.3.4巴西蘑菇菌絲體樣本製備及醱酵液的多醣體分離與純化 22
3.3.5巴西蘑菇醱酵液的多醣含量測定 22
3.3.6巴西蘑菇菌絲體之抗氧化性質測定 22
3.3.6.1抗氧化力之測定 22
3.3.6.2還原力測定 23
3.3.6.3捕捉1,1-diphenyl-2-picrylhydrazyl自由基能力測定 23
3.3.6.4螯合亞鐵離子之能力測定 24
3.3.6.5掃除超氧陰離子 24
3.4統計分析 25
第4章 結果與討論 26
4.1巴西蘑菇菌絲體固態培養 26
4.1.1巴西蘑菇菌絲體之平面生長速率 26
4.1.2巴西蘑菇菌絲體於不同脂肪酸下的生長情況 28
4.1.2.1巴西蘑菇菌絲體於YMA培養基添加不同脂肪酸(0.5
mg/ml)時的生長情況 29
4.1.2.2巴西蘑菇菌絲體於YMA培養基添加不同脂肪酸(1.0
mg/ml)時的生長情況 31
4.1.2.3巴西蘑菇菌絲體於YMA培養基添加不同脂肪酸(1.5
mg/ml)時的生長情況 33
4.1.2.4巴西蘑菇菌絲體於YMA培養基添加不同脂肪酸(2.0
mg/ml)時的生長情況 35
4.1.2.5巴西蘑菇菌絲體於YMA培養基添加不同脂肪酸(2.5
mg/ml)時的生長情況 37
4.2巴西蘑菇菌絲體搖瓶培養 39
4.2.1不同脂肪酸對搖瓶培養之巴西蘑菇菌絲體生長的影響 39
4.2.2不同脂肪酸對搖瓶培養之巴西蘑菇菌絲體多醣濃度的影響 40
4.3巴西蘑菇菌絲體醱酵槽培養 43
4.4巴西蘑菇菌絲體抗氧化能力測定 44
4.4.1抗氧化力 44
4.4.2還原力測定 48
4.4.3捕捉DPPH自由基能力測定 52
4.4.4螯合亞鐵離子能力測定 56
4.4.5掃除超氧陰離子能力測定 60
第5章 結論與建議 64
5.1結論 64
5.2建議 65
參考文獻 66
附錄 73
作者簡介 75
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