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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 蔣丙煌 | |
dc.contributor.author | Shiau-Wei Chen | en |
dc.contributor.author | 陳小薇 | zh_TW |
dc.date.accessioned | 2021-06-08T05:19:05Z | - |
dc.date.copyright | 2005-08-01 | |
dc.date.issued | 2005 | |
dc.date.submitted | 2005-07-28 | |
dc.identifier.citation | 六、參考文獻
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/24228 | - |
dc.description.abstract | 中文摘要
本實驗利用靈芝發酵特用植物 (氮源、碳源1),期望合併三者之保健效益,首先以 5 L 發酵槽探討發酵條件對發酵液成分之影響,繼而將初步所得發酵條件以 200 L 發酵槽放大量產。為了獲得更多發酵所產生之生理活性成分,乃以不同的發酵方法,以不同功能性成分為指標進行發酵試驗。最後以及時定量聚合酶鏈鎖反應偵測細胞激素 mRNA 的表現,並對靈芝發酵液進行抗過敏功能評估。 實驗結果顯示,含氮源 5%、碳源1 2%以及碳源2 2%之基質,在500 ml 之搖瓶試驗,以不調整 pH 值能獲得較高量之多醣及 (1→3)-β-D-glucan,於 5 L 發酵槽以不調整發酵培養基之 pH 值,探討接菌量對發酵過程中的影響,得知低接菌量之下的 (1→3)-β-D-glucan 的累積速度較快,而且對於三萜類的生成較有助益。因此,以不調整 pH 值,氮源 5%、碳源1 2%以及碳源2 2%,以 200 L 發酵槽放大量產,結果顯示大型發酵槽能得到更高量之多醣及 (1→3)-β-D-glucan。在兩階段式發酵及饋料式發酵中發現☆及溫度因子的限制有助於三萜類之生成,且饋料的方式能避免本身具抗菌成分之基質對靈芝生長的抑制,亦能獲得高量 (1→3)-β-D-glucan。 於抗過敏反應方面,靈芝發酵產品能抑制人類週邊血液單核細胞 (HPBMNCs) 細胞激素 IL-4、IL-5、IL-13 mRNA 表現,雖然對 IFN-γ、IL-2 mRNA 表現也有抑制情形出現,但是與 IL-4、IL-5、IL-13 mRNA 表現量相比,其抑制程度相對的較小,因而推論靈芝發酵液具有減輕過敏反應之功能。其中,又以氮源 5%及碳源2 2%在 x℃、通氣量 も vvm、もrpm 發酵 5 天,饋入碳源1 2%繼續發酵 6 天之發酵條件,所得之發酵產品最具有發展成抗過敏保健食品之潛力。 | zh_TW |
dc.description.abstract | Abstract
The medium consisting of B plants, C and A, was used for cultivating G. lucidum. It was found that cultivation of G. lucidum in 500 ml flask with the medium consisting of 5% C, 2% A ( A ), and 2% glucose yielded the highest amounts of polysaccharide and (1→3)-β-D-glucan. Low inoculation density could promote the production of (1→3)-β-D-glucan and crude triterpenoids during submerged fermentation at x℃, aeration rates も vvm, and agitation speed もrpm in a 200L fermenter. Due to the environment and temperature restriction, the two-stage fermentation could improve crude triterpenoids production. And fed batch cultivation could further increase the yield of polysaccharide and (1→3)-β-D-glucan in the fermentation broth. For the anti-allergic effect, the mononuclear cells (MNCs) from human was used. Results indicated that the fermented products could reduce the mRNA expression of the TH1 cytokines IL-4, IL-5, IL-13 by the mitogen-activated MNCs. Although the fermented products also reduced the mRNA expression of the TH2 cytokines IFN-γand IL-2, the extend of reduction was less than that mRNA expression of the TH1 cytokines. | en |
dc.description.provenance | Made available in DSpace on 2021-06-08T05:19:05Z (GMT). No. of bitstreams: 1 ntu-94-R92641007-1.pdf: 1063444 bytes, checksum: 1d053d408b23dbaf19e9954edcf005e8 (MD5) Previous issue date: 2005 | en |
dc.description.tableofcontents | 目 次
中文摘要…………………………………………………...……………..I 英文摘要………………………………………………...……………….II 前言…………………………………………………………………..... 1 壹、文獻回顧 一、靈芝之簡介 (一) 靈芝分類…………………………………………………… ..3 (二) 靈芝活性成分 (1) 多醣類…………………………………………….………..4 (2) 活性多醣 (1→3)-β-D-glucan……………………..………5 (3) 三萜類物質………………………………………………. 6 (三) 靈芝栽培方式 (1) 野生靈芝…………………………………………………..8 (2) 人工栽培靈芝……………………………………………..9 (四) 深層液態醱酵 (1) 培養基成分……………………………………………….10 (2) 培養條件……………………...…………………………..13 (五) 不同之醱酵方式 (1) 兩階段式醱酵…………………...……………………… 15 (2) 饋料式醱酵……………………………………………… 16 二、特用植物基質之簡介 三、過敏反應 (Hypersensitivity or Allergy) (一) 過敏反應之簡介……………………………………………..20 (二) 細胞激素之功能………………………….………….………22 (三) TH1 和 TH2 細胞亞群與平衡………..…………………….23 四、聚合酶鏈鎖反應與即時定量聚合酶鏈鎖反應 (一) 聚合酶鏈鎖反應 (PCR)…………………………………….24 (二) 即時定量聚合酶鏈鎖反應 (1)即時螢光定量 PCR 技術的原理與方法……….………...26 (2)螢光化學……………………………………….…………27 (3) 定量方法…………………………………..…………….31 (4) 即時螢光定量PCR技術的應用………………………...32 貳、實驗架構…………………………………………………………...34 參、材料方法 一、Ganoderma lucidum 醱酵特用植物之試驗…….………………37 (一) 實驗材料 (1) 菌株來源………………………..…………………….….. 37 (2) 藥品……………………………….……………………….37 (3) 儀器………………………………………………………..39 (二) 實驗方法 (1) 菌株的培養………………………………………………40 (2) 三角瓶培養………………………………………………41 (3) 接菌量……………………………………………………41 (4) 醱酵………………………………………………………41 (5) 培養基製備………………………………………………42 (6) 分析方法…………………………………………………43 (7) 醱酵條件…………………………………………………46 二、探討靈芝醱酵產物對過敏反應之影響 (一) 實驗材料 (1) 血液來源……………………….………………………….48 (2) 樣品…………………………….………….………………48 (3) 藥品…………………………….…………………….……48 (4) 器材…………….…………………….……………………49 (5) 儀器……………………………….……….………………50 (二) 實驗方法 (1) 人類周邊血球細胞分離…………..………………………51 (2) 靈芝醱酵液對於人類周邊血球細胞增生測定方法..……52 (3) 人類周邊血液單核細胞之活化……………….………….53 (4) 人類周邊血液單核細胞 RNA 之萃取………………….53 (5) RNA 的測量……………………………….……………54 (6) 反轉錄反應……………………………….………………54 (7) 即時定量聚合酶連鎖反應………………………….……55 (8) 基因之引子序列設計……………….…………….………57 (9) 統計分析…………………………………………….……57 肆、結果討論 第一部分:以特用植物當基質之靈芝醱酵條件之探討 一、起始pH對靈芝醱酵特用植物活性成分之影響……………..59 二、接菌量對靈芝醱酵液活性成分之影……………………..…..64 (一)、接菌量對靈芝生長情形之影響……………………………..64 (二)、接菌量對於靈芝醱酵過程中總多醣與(1→3)-β-D-glucan生成量之影響………..…………….………………………….69 (三)、接菌量對於靈芝醱酵過程中三萜類生成量之影響……......71 三、200公升醱酵槽之醱酵試驗……………………………...….. 74 (一)、以200 L醱酵槽放大量產對於靈芝生長情形之觀察……...74 (二)、以200 L醱酵槽放大量產靈芝於醱酵過程中總多醣與 (1→3)-β-D-glucan生成量之變化……………………..…77 第二部分:不同醱酵方式對靈芝醱酵產品之活性成分影響 一、兩階段式之醱酵試驗(Two-stage cultivation)…………….. 83 (一)、兩階段式之醱酵對於靈芝生長情形之觀察……………..…83 (二)、兩階段式之醱酵對於靈芝醱酵中總多醣與(1→3)-β-D-glucan生成量之影 響……………………….……….….83 (三)、兩階段式之醱酵對於靈芝醱酵過程指標性成分粗三萜生成量之影響………………………….………………………….84 二、饋料式之醱酵試驗(Fed-batch cultivation)……………….….88 (一)、以饋料方式醱酵對於靈芝生長情形之觀察…………..……88 (二)、以饋料方式醱酵對於靈芝醱酵過程指標性成分總多醣與 (1→3)-β-D-glucan 生成量之影響……………………..92 (三)、以饋料方式醱酵對於靈芝醱酵過程中 粗三萜生成量之影響……………………………………..92 第三部分:靈芝醱酵產物對過敏反應之影響 (一)、靈芝醱酵樣品對人類周邊血液單核細胞 (HPBMNCs)生長能力之影響……………………………..……………..…..99 (二)、靈芝醱酵樣品對人類周邊血液單核細胞 (HPBMNCs)分泌細胞激素mRNA表現之影響………………………...….100 伍、結論………………………………………………………………..111 陸、參考文獻………………………………………..…………………113 圖 次 圖一、靈芝 ( Ganoderma lucidum (Leyss ex Fr) Karst)之子實體…….2 圖二、(1→3)-β-D-glucan之結構………………………………………..6 圖三、三萜類成分之結構……………………...……………………….. 7 圖四、四種經由免疫機制產生之過敏反應……………………………20 圖五、即時定量PCR之螢光化學標定法 (A) SYBR Green 1 (B) Taqman (C) molecular beacons………………………………30 圖六、培養基起始pH值對靈芝於500 ml三角瓶培養時總多醣生成量之影響………………………………….…………………….61 圖七、培養基起始pH值對靈芝於500 ml三角瓶培養時(1→3)-β-D-glucan生成量之影響…………………………………..…62 圖八、培養基起始pH值對靈芝於500 ml三角瓶培養時 crude triterpenoids生成量之影響……………...……………….63 圖九、不同接菌量對靈芝醱酵液pH值之變化情形…………………..65 圖十、不同接菌量對靈芝醱酵液中還原醣之變化情形……..………..67 圖十一、不同接菌量對靈芝醱酵過程中菌絲球粒徑之變化情形...….68 圖十二、不同接菌量對靈芝醱酵液中總多醣之變化…………...…….70 圖十三、不同接菌量對靈芝醱酵液中(1→3)-β-D-glucan之變化 情形…………………………….…………………………….72 圖十四、比較200L與5L醱酵槽對靈芝醱酵過程中pH值之變化情形………………………………………………………….….75 圖十五、比較200L與5L醱酵槽對靈芝醱酵過程中還原醣之變化情形…………………………………………………………….76 圖十六、比較200L與5L醱酵槽對靈芝醱酵過程中總多醣之變化情形………………………………………………………...…..78 圖十七、比較200L與5L醱酵槽對靈芝醱酵過程中(1→3)-β-D-glucan 之變化情形…………………………………….…………….79 圖十八、兩階段式醱酵對靈芝醱酵過程中pH.………………………..81 圖十九、兩階段式醱酵對靈芝醱酵過程中還原醣之變化情形………82 圖二十、兩階段式醱酵對靈芝醱酵過程中總多醣之變化情形……....85 圖二十一、兩階段式醱酵對靈芝醱酵過程中(1→3)-β-D-glucan之變化情形….…………………………………………………….86 圖二十二、饋料式醱酵對靈芝醱酵過程中pH之變化情形……………………………………………………………..89 圖二十三、饋料式對靈芝醱酵過程中菌絲球粒徑之變化情形……………………………………………………………. 90 圖二十四、饋料式醱酵對靈芝醱酵過程中還原醣之變化情形…………………………………………….……………….91 圖二十五、饋料式醱酵對靈芝醱酵過程中總多醣之變化情形……………………………………………….…………….93 圖二十六、饋料式醱酵對靈芝醱酵過程中(1→3)-β-D-glucan之變化情形………………………………………………………… 94 圖二十七、比較不同醱酵方式靈芝醱酵至第も 天時功能性成分的含量……………………………………………………….…….95 圖二十八、各種培養方式生產之靈芝醱酵液對PBMNCs中IL-4 mRNA表現的影響……….…………………………………..…….106 圖二十九、各種培養方式生產之靈芝醱酵液對PBMNCs中IL-5 mRNA表現的影響……………...………………………………….107 圖三十、各種培養方式生產之靈芝醱酵液對PBMNCs中IL-13 mRNA表現的影響…………..….………………………………….108 圖三十一、各種培養方式生產之靈芝醱酵液對PBMNCs中IL-2 mRNA表現的影響……………….……………………………….109 圖三十二、各種培養方式生產之靈芝醱酵液對PBMNCs中IFN-γ mRNA表現的影響………...……………………………….110 表 次 表一、本實驗所設計細胞激素IL-2、IL-4、IL5、IL13、INF-γ | |
dc.language.iso | zh-TW | |
dc.title | 各種液態培養靈芝方式對產品之活性成分及抗過敏功能之影響 | zh_TW |
dc.title | Submerged Cultivation of Ganoderma lucidum for the Production of Bioactive Components and the Anti-allergic Function of the Fermentation Products | en |
dc.type | Thesis | |
dc.date.schoolyear | 93-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 孫璐西,徐敬衡,繆希椿,沈立言 | |
dc.subject.keyword | 靈芝,深層液態醱酵, | zh_TW |
dc.subject.keyword | Ganoderma lucidum,submerged fermentation, | en |
dc.relation.page | 126 | |
dc.rights.note | 未授權 | |
dc.date.accepted | 2005-07-29 | |
dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
dc.contributor.author-dept | 食品科技研究所 | zh_TW |
顯示於系所單位: | 食品科技研究所 |
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