臺灣博碩士論文加值系統

本論文探討牛樟芝Antrodia camphorata NPU-44菌絲體固態發酵多醣體結構特徵與其免疫調節之效應，本研究內容正申請專利中，暫不公開。

摘要……………………………………………………………………… I
Abstract…………………………………………………………………… III
謝誌……………………………………………………………………… V
目錄……………………………………………………………………… VI
圖表目錄…………………………………………………………………. X
壹、 前言……………………………………………………………… 1
貳、 文獻回顧…………………………………………………………. 4
一、樟芝簡介…………………………………………………… 4
(一) 牛樟芝子實體與菌絲體型態…………………………. 4
(二) 牛樟芝命名史………………………………………… 4
(三) 牛樟芝菌種鑑定…………………………………........ 6
二、牛樟芝機能性成分及其生物活性…………………………. 6
(一) 固醇類及其生物活性………………………………… 6
(二) 三萜類及其生物活性…………………………………. 7
(三)核苷類及其生物活性…………………………………... 7
(四)多醣體及其生物活性…………………………………... 8
(五)其他生物活性…………………………………………... 8
1. 抗氧化活性………………………………………….. 8
2. 護肝生物活性……………………………………….. 8
三、多醣體之功能性機制………………………………………. 9
(ㄧ) 多醣體免疫活性機制…………………………………. 9
(二) 多醣體結構與其免疫活性之關連……………………. 10
四、牛樟芝菌絲體多醣體結構特徵…………………………….. 14
五、多醣體之結構分析原理…………………………………….. 14
(ㄧ) 利用高效能液相層析儀測定分子量之原理與應用………………………………………………………
14
(二) 利用氣相層析儀測定單醣組成之原理與應用……… 15
(三) 利用氣相層析質譜儀測定醣苷鍵結之原理與應用… 15
(四) 利用傅立葉紅外線光譜測定官能基團之原理與應用………………………………………………………
16
(五)利用核磁共振光譜儀鑑定結構之原理與應用……… 16
參、 材料與方法…………………………………………………......... 22
一、實驗設計…………………………………………………….. 22
二、材料與方法………………………………………………….. 23
(一) 材料……………………………………………………... 23
(二) 分離菌株………………………………………………... 23
1. 平板培養基配製…………………………………….. 23
2. 液態培養基配製…………………………………….. 23
3. 菌種分離…………………………………………….. 24
(三)樟芝固態發酵……………………………………………. 24
1. 胚芽米固態培養基配製…………………………….. 24
2. 接種菌株…………………………………………….. 24
(四)菌株鑑定…………………………………………………. 25
1. 萃取純化DNA………………………………………. 25
2. 聚合酶連鎖反應…………………………………….. 25
3. 電泳膠分析………………………………………… 25
(五)萃取樟芝固態發酵物之粗多醣體…………………....... 26
(六)去鹽…………………………………………………....... 26
(七)多醣體濃度的測定…………………………………....... 27
1. 標準品……………………………………………… 27
2. 粗多醣樣品………………………………………… 27
(八)分離純化粗多醣體……………………………………... 27
(九)分子量測定………………………………………..……. 27
1. 標準曲線…………………………………………… 27
2. 樣品分析…………………………………………… 28
(十)單糖組成分析………………………………………....... 28
(十一)多醣體鍵結分析……………………………...……… 29
(十二)官能基分析………………………………………....... 29
(十三)結構鑑定…………………………………………....... 30
(十四)免疫力測試………………………………………....... 30
1. 實驗動物…………………………………………… 30
2. 採集老鼠血液及脾臟……………………………… 30
3. 周邊血液單核細胞分離…………………………… 30
4. 脾臟細胞分離……………………………………… 30
5. 總免疫細胞增殖作用……………………………… 31
(十五) 製備抗ACP-179之多株抗體……………………… 31
1. 實驗動物…………………………………………… 31
2. 免疫兔子…………………………………………… 31
3. 間接性酵素連結免疫吸附分析法………………… 32
4. 免疫擴散法………………………………………… 32
肆、 結果與討論………………………………………………………. 34
ㄧ、菌種鑑定…………………………………………………… 34
二、牛樟芝菌絲體固態發酵及其粗多醣體製備……………… 34
三、分離及純化牛樟芝菌絲體固態發酵物之粗多醣體……… 41
(一) 分離及純化粗多醣體…………………………………. 41
(二) 多醣體分子量…………………………………………. 45
四、總免疫細胞增殖…………………………………………….. 51
五、多醣體的單糖組成………………………………………….. 56
六、多醣體鍵結………………………………………………….. 56
七、官能基分析………………………………………………….. 62
八、結構鑑定……………………………………………………. 67
九、間接性酵素連結免疫吸附及免疫擴散測試………………. 71
伍、 結論…………………………………………………………......... 80
參考文獻…………………………………………………………………. 81
作者簡介…………………………………………………………………. 88

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