臺灣博碩士論文加值系統

烏苓參(Xylaria nigripes)又名黑柄炭角菌，屬於炭角菌科(Xylariaceae)真菌，為一種珍貴的藥用菇菌。依據中國民間藥典之記載，烏苓參具有利尿、補心腎、增強免疫、改善失眠、調理婦女生理及改善產後失血症等功效，但這些療效仍停留於民間傳說。為了發展烏苓參產業，了解其培養條件及量產技術為目前研究重點之一。本研究的主要目的為利用液態基質進行探討三株不同烏苓參菌株(XN-1、XN-2 及XN-3)在不同培養條件下之菌絲體生長速率及產量，同時評估其多醣體產量及DPPH 自由基之清除效果。實驗結果顯示，於基本培養基K2HPO4、KH2PO4、MgSO4 • 7H2O 0.05%下，XN-1 之最佳培養條件為glucose 2.5%，yeast extract 0.5 %、溫度30℃、振盪速度250 rpm 及pH 8.0培養7天，其菌絲體產量及多醣產量分別為12.2±0.01 g/L及10.17±0.09%；XN-2 之最佳培養條件為fructose 2.5%、peptone 0.3%、溫度26℃、振盪速度150 rpm 及pH 7.0培養8天，菌絲體產量及多醣產量分別為10.9±0.01 g/L及10.22±0.06%；XN-3 之最佳培養條件為玉米粉5%、黃豆粉0.5%、溫度22℃，振盪速度200 rpm 及pH 6.0培養8天，其菌絲體產量及多醣產量分別為11.3±0.01 g/L及9.94±0.10%。此最適條件培養之XN-1、XN-2 及XN-3 菌絲體產量與多醣體產量皆比市售PDB (potato dextrose broth)培養基所得到的結果高一倍，在清除DPPH 自由基能力，最適培養基之菌絲體水粗萃取物均比以PDB 培養基培養萃取物效果強。綜合以上結果說明，烏苓參菌絲體之產量與多醣體產量可於最適條件培養下提高，同時亦可提高其清除DPPH 自由基能力。

Xylaria nigripes (Koltz.) Sacc., also known as Wu Ling Shen, is a high value medicinal fungus of family Xylariaceae. According to the traditional Chinese pharmacopoeia, it possesses the properties of diuretic, invigorating kidney and enhancing immune function, as well as improving insomnia, woman menstrual cycle and bleeding after giving birth. In order to develop the Xylaria nigripes industry, it is important to understand the conditions and technique for its large scale production. The present study aimed to examine the mycelial growth rate and production of three different Xylaria nigripes (XN-1, XN-2 and XN-3) isolates using liquid fermentation. In addition, their polysaccharides content and DPPH radical scavenging activity were also evaluated. Results showed that under the basic medium comprising K2HPO4、KH2PO4、MgSO4 • 7H2O 0.05%, the optimal cultured condition for XN-1 was glucose 2.5%, yeast extract 0.5 %, 30℃, shaking speed 250rpm, pH 8.0 in 7 days, of with the mycelium weight and polysaccharide content were 12.2±0.01g/L and 10.17±0.09% respectively. For XN-2, the best condition was fructose 2.5%, peptone 0.3%, 26℃, shaking rate 150 rpm, pH 7.0 in 8 days, with the mycelial weight and polysaccharide content 10.9±0.01 g/L and 10.22±0.06%, respectively. For XN-3, the optimal condition was corn powder 5%, soya bean powder 0.5%, 22℃, shaking rate 200rpm, pH 6.0 in 8 days, with the mycelial weight and polysaccharide content 11.3±0.01 g/L and 9.94±0.10%, respectively. Under this optimal condition , XN-1, XN-2 and XN-3 exhibitd a better mycelial growth and polysaccharide production than PDB. Extracts of these mycelium also showed a better DPPH radical scavenging activity than that of PDB culture. Take together, these results conclude that the growth rate and polysaccharide production of XN can be improved by optimizing the condition of liquid fermentation, which can also enhance the DPPH radical scavenging activity.

中文摘要…………………………………………………………………. Ⅰ
英文摘要…………………………………………………………………. Ⅲ
誌謝………………………………………………………………………. Ⅴ
目錄………………………………………………………………………. Ⅹ
圖次………………………………………………………………………. XI
表次………………………………………………………………………. XII
第壹章 緒論……………………………………………………………... 1
1.1 前言……………………………………………………………….. 1
1.2 研究目的………………………………………………………….. 2
1.3 研究動機………………………………………………………….. 2
第貳章 文獻回顧………………………………………………………... 3
2.1 烏苓參的研究簡史……………………………………………….. 3
2.2烏苓參的分類地位及生活史………………………………….... 3
2.3 烏苓參的外觀…………………………………………………….. 4
2.4 烏苓參之活性物質及藥理研究………………………………….. 5
2.5 烏苓參生長所需基質探討……………………………………….. 11
第參章 研究架構………………………………………………………... 16
第肆章 材料與方法…………………………………………………….. 17
4.1 實驗菌株…………………………………………………………. 17
4.2 實驗藥品…………………………………………………………. 17
4.3 實驗器材…………………………………………………………. 17
4.4 實驗設備…………………………………………………………. 18
4.5 實驗方法………………………………………………………….. 18
4.5.1建立最適培養條件…………………………………………… 18
4.5.1.1菌種增殖……………………………………………………. 18
4.5.1.2菌種活化保存………………………………………………. 18
4.5.2 種菌菌液之製備……………………………………………... 18
4.5.3 培養溫度…………………………………………………….. 19
4.5.4 培養時間…………………………………………………….. 19
4.5.50 pH…………………………………………………………… 19
4.5.6 振盪速度…………………………………………………….. 19
4.5.7 碳源種類…………………………………………………….. 20
4.5.8 氮源種類…………………………………………………….. 20
4.5.3多醣含量分析………………………………………………… 21
4.5.3.1多醣檢量線…………………………………………………. 21
4.5.3.2 酵素透析………………………………………………….... 21
4.5.4 DPPH自由基清除活性試驗…………………………………. 22
4.5.4 統計分析……………………………………………………... 23
第伍章 結果……………………………………………………………... 24
5.1 烏苓參之最適培養溫度……………………………………….. 24
5.1.2不同溫度培養對烏苓參胞外多醣體產量之影響…………… 24
5.2烏苓參之最適培養時間………………………………………… 25
5.2.1不同培養時間對烏苓參胞外多醣體產量之影響…………… 25
5.3烏苓參之最適培養pH值………………………………………. 26
5.3.1不同培養pH值對烏苓參胞外多醣體產量之影響………….. 26
5.4烏苓參之最適培養振盪速度…………………………………… 27
5.4.1不同培養振盪速度對烏苓參胞外多醣體產量之影響………. 27
5.5烏苓參之最適培養碳源………………………………………… 28
5.5.1不同培養碳源對烏苓參胞外多醣體產量之影響……………. 28
5.6烏苓參之最適培養氮源………………………………………… 29
5.6.1不同培養氮源對烏苓參胞外多醣體產量之影響……………. 29
5.7最適培養條件與PDB 培養基培養之烏苓參菌絲體產量比較. 30
5.7.1最適培養基與PDB 培養基培養之胞外多醣體產量……….. 30
5.8最適培養條件與PDB 培養基培養之烏苓參菌絲體萃取物的
DPPH自由基清除活性................................................................. 31
第陸章 討論……………………………………………………………... 32
第柒章 結論……………………………………………………………... 39
參考文獻……………………………………………………………......... 41
附錄………………………………………………………………………. 49
作者簡介…………………………………………………………………. 68

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