臺灣博碩士論文加值系統

鬼傘屬真菌主要特色是生長週期短，子實體成熟後菌蓋會自溶呈墨汁狀，本研究重點著重於探討不同條件下，鬼傘屬真菌菌絲體自溶進行程度之差異。首先利用液態培養方式挑選生長迅速之菌株，經七天液態搖瓶培養後，Coprnius cinereus BCRC 36099生長最為迅速，第七天菌絲體乾菌體量可達1.215(g/100ml 培養基)。
在不同溫度與緩衝溶液pH值對真菌自體溶解影響試驗中，當pH值=6、溫度=50℃、反應24小時，自體溶解率達66.92%。在時間因素對真菌自體溶解影響試驗中，在第1小時自體溶解率已達到60.16%，至第24小時，自體溶解率僅上升了6.76%。在添加不同酵素對真菌自體溶解影響試驗中，添加木瓜酵素自體溶解率為69.99%，添加鳳梨酵素自體溶解率為68.06%。
當pH值=6、溫度=50℃、反應24小時，鳳梨酵素對於幾丁質與蛋白質的分解成效最佳，反應後得葡萄糖氨與游離胺基酸濃度分別是85.7835μg/ml與1.1894mg/ml。另外，在探討發酵槽自體溶解試驗中，菌絲體濃度減少39.5%，葡萄糖氨與游離胺基酸的濃度分別是270.7550 (μg/ml)與5.3519(mg/ml)。

The character of fungi Coprinus is the short growing periods. The cap of the fruit body will autolysis and be ink-like after maturity. This research focuses on the effects of Corprnius mycelial autolysis degree under different reaction conditions. By using a liquid culture method, we select the strain in which growth is the most rapid。After seven days of flask liquid cultivation, the Coprnius cinereus BCRC 36099 exhibited the most rapid growth, and based on the dry weight, the mycelium mass can be up to 1.215(g/100ml substrate).
The optimum condition for fungi autolysis was determined to be a temperature equal to 50℃. Initial pH=6 was controlled before reaction. The autolysis solubility can reach to 60.16% at 1st hour, and 66.92% after 24 hours of reaction in 0.1M sodium acetate buffer. In addition, adding 0.1g of the papain and bromelain can bring the autolysis solubility up to 69.99% and 68.06%.
During the reaction in 24 hours, the addition of bromelain can obtain the best dissolution result for chitin and protein; the concentration of glucosamine and free amino acids after reaction are 85.7835(μg/mL) and 1.1894(mg/mL). Moreover, in the fermenter operation for autolysis reaction, mycelium concentration decreased 39.5%, glucosamine concentration and free amino acids concentration are 270.7550 (μg/mL) and 5.3519(mg/mL) individually.

中文摘要 ..............................................Ⅰ
英文摘要...............................................Ⅱ
誌謝...................................................Ⅲ
目錄...................................................Ⅳ
圖目錄.................................................Ⅷ
表目錄.................................................Ⅸ

第一章 緒論.............................................1
1-1 前言................................................1
1-2 本文大綱............................................3

第二章 文獻回顧.........................................4
2-1 食用菇類之營養價值..................................4
2-2 食用菇類菌絲體培養物之利用..........................15
2-3 自體溶解(Autolysis).................................19
2-4 鬼傘屬真菌..........................................24

第三章 實驗材料與分析方法...............................28
3-1 菌株................................................28
3-2 實驗藥品............................................29
3-3 實驗儀器及設備......................................30
3-4 培養基組成..........................................31
3-4-1 斜面與培養皿培養基：............................31
3-4-2 液態培養基......................................31
3-5 分析方法............................................32
3-5-1 pH 值測定.......................................32
3-5-2 搖瓶培養之菌體乾重測定..........................32
3-5-3 鬼傘真菌自體溶解度之測定........................32
3-5-4 總醣之定量分析..................................33
3-5-5 蛋白質之定量分析................................35
3-5-6 游離胺基酸之定量分析............................38
3-5-7 葡萄糖氨之定量分析..............................41

第四章 實驗方法.........................................43
4-1 平面培養............................................43
4-1-1 試管斜面培養....................................43
4-1-2 培養皿培養......................................44
4-1-3 切割器製作方法..................................45
4-2 液態培養............................................46
4-2-1 種菌的製備......................................46
4-2-2 搖瓶液態培養....................................47
4-2-3 發酵槽液態培養..................................48
4-3 菌種挑選............................................49
4-3-1 平面培養皿生長挑選..............................49
4-3-2 搖瓶液態培養的挑選..............................50
4-4 鬼傘屬真菌自體分解實驗..............................51
4-4-1 均質液的製備....................................51
4-4-2 溫度與緩衝溶液pH值對真菌自體溶解影響試驗.......52
4-4-3 時間對真菌自體溶解影響試驗(24 小時) ...........53
4-4-4 時間對真菌自體溶解影響試驗(1 小時).............54
4-4-5 添加不同酵素對真菌自體溶解影響試驗.............55
4-4-6 自體溶解液中總醣濃度之變化.....................56
4-4-7 自體溶解液中葡萄糖氨濃度之變化.................57
4-4-8 自體溶解液中蛋白質濃度之變化...................58
4-4-9 自體溶解液中游離胺基酸濃度之變化...............59
4-4-10 探討攪拌式發酵槽進行自體溶解試驗..............60
4-4-11溫度與緩衝溶液pH值對不同酵素分解蛋白質活性測試
..............................................61

第五章 結果與討論 ......................................62
5-1 菌種挑選...........................................62
5-1-1 平面培養皿生長挑選.............................62
5-1-2 搖瓶液態培養的挑選.............................65
5-2 鬼傘屬真菌自體溶解實驗.............................67
5-2-1 溫度與緩衝溶液pH值對真菌自體溶解影響試驗......67
5-2-2 時間對真菌自體溶解影響試驗(24 小時)............69
5-2-3 時間對真菌自體溶解影響試驗(1 小時).............71
5-2-4 添加不同酵素對真菌自體溶解影響試驗.............73
5-2-5 自體溶解液中總醣濃度與葡萄糖氨濃度之變化情形...75
5-2-6 自體溶解液中蛋白質濃度與游離胺基酸濃度之變化情形
...............................................79
5-2-7 探討攪拌式發酵槽進行自體溶解試驗...............83
5-2-8 溫度與緩衝溶液pH值對不同酵素分解蛋白質活性測試
...............................................85
第六章 結論及未來展望..................................87
6-1 結論...............................................87
6-2 未來展望...........................................89

參考文獻...............................................90

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