臺灣博碩士論文加值系統

羊肚菌(Morchella esculenta)是著名的珍貴野生食用菌，目前子實體難以人工培養。由於一般菇類菌絲體經由液態發培養時，在定速攪拌下，菌體聚集效應明顯，致使有效種菌菌絲量迅速降低且造成養份質傳上的困難。本研究重點在於探討不同的攪拌策略對羊肚菌液態培養菌絲體生長之影響與自體分解研究。

在發酵槽中的分析，證明羊肚菌菌絲為凝聚型，發酵初期以定速攪拌會使菌體濃度迅速下降，於40 hr取樣菌體濃度趨近於0 g/L。若以間歇式攪拌可以減緩菌絲凝聚趨勢，提升菌體濃度。而在實驗進行48 hr之後用不同的攪拌策略中，若只以定速攪拌，則菌體容易凝聚造成菌體濃度下降，因此最後以定速攪拌300rpm伴隨500rpm (1min/8hr)的攪拌於152hr可得最佳菌重7.63g/L。若發酵槽配合連續式均質機，則可防止菌絲球型化，加快其生長速度而提升產率，88hr即可得7.73 g/L。

而在經發酵槽培養後，羊肚菌會因攪拌操作策略的不同而顯現不同自體分解能力，在取樣於菌體凝聚而濃度下降時的菌體較容易自體分解，於碳氮充足情況下取樣則反之。在不添加其他酵素情況下以25oC，24hr下菌體即可達到最佳自體分解，若需添加酵素，則以木瓜酵素0.1%，70oC，pH=7狀況下有最佳水解能力。

Morchella esculenta is one of the most valuable edible mushrooms, which still could not be cultivated easily. During the culture of filamentous fungi, the phenomenon of rapid aggregation causes the decrease of amount of dispersed mycelia and the formation of pellets will lead to the problem of diffusion limitation. Due to the fact that large pellets often occurs in the mycelium submerged culture of Morchella esculenta, the effect of different agitation schemes on mycelium aggregation was investigated in 5L fermentor in this study. A novel scheme using intermittent agitation could maintain a higher level of mycelium fragment concentration for 48 hours and enhanced the biomass concentration to the level of 7.63 g/L at 152hr. When the fermentor was connected with a homogenizer, mechanical shear force could reduce the pellet size and numbers, but also damaged the mycelium at the same time. However, the biomass concentration rose to 7.73 g/L at 88hr.
The results also demonstrated that the aggregated mycelia or the mycelia from the culture connected with a homogenizer tended to be easy for autolysis. In contrast, high amount of carbon and nitrogen source might resist the occurrence of autolysis. The optimum conditions for the autolysis of Morchella esculenta was found to be around 25oC in 24 hr. The addition of 0.1% papain could enhance the autolysis under the conditions of 70oC and pH 7.

Keyword：Morchella esculenta, filamentous fungi morphology, intermittent agitation, autolysis.

目 錄
中文摘要 ………..…………………………………………………………….I
英文摘要 ………..…………………….……………………………………..III
目錄……………………………………………………………………………V
圖目錄……………………………………………………….……………...VIII
表目錄 ……………..…………………………………………………………X

第一章 緒論……..……………………………………………………...…1
1-1羊肚菌簡介…………………………………………………………………2
1-2羊肚菌的生長環境………………………………………………………....6

第二章 文獻回顧 ...……...……………………………………..….……7
2-1 羊肚菌的液態培養…………………………………………………..……8
2-2 絲狀真菌之生物特性…………………………………………………....18
2-3 絲狀真菌型態之影響因子…………………………………….………...20
2-4 絲狀真菌培養之生物反應器設計……………………………................29
2-5 絲狀真菌的自體分解…………………………………………………....32

第三章 實驗材料與分析方法………………………………………...42
3-1 菌株……………………………………………………………….……...42
3-2 實驗藥品 ………………………………………………………….….…43
3-3 實驗儀器與設備 …………………………………………………..……45
3-4 實驗方法……………………………………………………………..…..47
3-5 分析方法 …………………………………………………………..……50

第四章 液態培養下的菌種篩選與培養基改良…………………….51
4-1 前言…………………………………………………………………..…..51
4-2液態培養下的菌種篩選實驗………………………………...………..…52
4-3培養基改良實驗 ……………………………………………………..….55
4-4 結論………………………………………………………………………58

第五章 定速攪拌對羊肚菌之影響….…………………………...…. 59
5-1 前言……………………………………………………………………...59
5-2定速攪拌對羊肚菌之影響……………………………………...…….…60
5-3 結論……………………………………………………………………...64

第六章 羊肚菌於不同的間歇攪拌操作之發酵槽比較………….65
6-1 前言……………………………………………………..………….……65
6-2羊肚菌菌體於發酵槽中之間歇攪拌實驗A…………..………………..66
6-3羊肚菌菌體於發酵槽中之間歇攪拌實驗B……...……………….……71
6-4羊肚菌菌體於發酵槽中之間歇攪拌實驗C……...……………….……76
6-5結論………………………………………………….………………...…80

第七章 羊肚菌於發酵槽之間歇攪拌與連續式均質機的搭配操作.....81
7-1 前言……………………………………………………..…………….…81
7-2間歇攪拌與連續式均質機的搭配操作實驗D....…..……………..……82
7-3間歇攪拌與連續式均質機的搭配操作實驗E.…..……………….……86
7-4間歇攪拌與連續式均質機的搭配操作實驗F. …..………….…………90
7-5間歇攪拌與連續式均質機的搭配操作實驗G. …..………….…………95
7-6結論………………………………………………….………….………..99

第八章 羊肚菌的自體分解測試………………………….………….101
8-1 前言………………………………………...………………………..….101
8-2不同溫度與pH值下的自體分解……………………………………….102
8-3不同酵素添加於羊肚菌的分解實驗……………...……………..……..104
8-4不同酵素添加於羊肚菌的分解實驗(續) ……………...……….………107
8-5綜合討論……….………………………………………………….….…109

第九章 結論與未來展望………………………………...…....…….…110
9-1 結論………………………………………………………………..……110
9-2 未來展望……………………………………………………………..…111

參考文獻……………………………………………………………………. 112

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