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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 蔣丙煌(Been-Huang Chiang) | |
dc.contributor.author | Yi-Ling Wu | en |
dc.contributor.author | 鄔宜伶 | zh_TW |
dc.date.accessioned | 2021-06-15T11:36:52Z | - |
dc.date.available | 2025-08-16 | |
dc.date.copyright | 2020-08-20 | |
dc.date.issued | 2020 | |
dc.date.submitted | 2020-08-16 | |
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/49598 | - |
dc.description.abstract | 牛樟芝 (Antrodia cinnamomea) 是台灣原生種真菌,其液態發酵產出之菌絲體含有高量之具有抗癌功效的泛醌類 antroquinonol (AQ) 及 4-acetylantroquinonol B (4-AAQB) ,其中 4-AAQB是菌絲體中主要之抗肝癌化合物。本研究室過去曾利用超音波作為外力刺激,發現可有效促進液態發酵牛樟芝菌絲體中 4-AAQB 生合成,但是不能刺激 AQ 的生合成。本研究再次證實此一結果的正確性,而其原因可能是由於生合成途徑中的酵素活性受超音波改變所致。本研究原本預期,在可促進 4-AAQB 生合成的超音波處理條件下,牛樟芝應能更有效地運用六碳環前驅物 CoQ0 生合成 4-AAQB ,但結果卻不如預期,在給予超音波刺激下,外加的 CoQ0 無法有效被用於 4-AAQB 生合成。由 LC-MS-MS 分析則可推論, CoQ0 應可經由 polyketide pathway 被代謝為 CoQ3 與 CoQ3B ,故 CoQ0 應同時是 AQ 與 4-AAQB 的六碳環前驅物,而添加 CoQ0 無法提升 4-AAQB 產量可能是由於菌絲體中累積的 CoQ3B 與 AQB 無法進一步被轉為 4-AAQB 所致。此外,本研究也推論,超音波處理對 polyketide pathway 的主要影響,是能促進其中間產物 5-DMQ3B 轉為 CoQ3B 的步驟,並能抑制其下游產物 CoQ3 轉為 AQ 之步驟。若同時添加 CoQ0 並使用超音波處理,超音波的介入則可能會使部分外加的 CoQ0 被轉而用於其他代謝途徑而造成 CoQ3B 生合成減少,並可能抑制 5-DMQ3 與 6-DMQ3 轉為 CoQ3 之步驟,進而抑制 4-AAQB 及 AQ 的生合成。 | zh_TW |
dc.description.abstract | Antrodia cinnamonea is an endemic fungus of Taiwan that specifically inhabits the decayed inner wall of stout camphor trees. The mycelia of A. cinnamonea produced through submerged fermentation are rich in anti-hepatic compounds such as antroquinonol (AQ) and 4-acetylantroquinonol B (4-AAQB), and the latter has been proven to possess the most potent anti-hepatocellular carcinoma function. Our laboratory has used ultrasound as an external force to promote 4-AAQB biosynthesis, and this study further confirmed that ultrasound treatment indeed could increase 4-AAQB production in the mycelia, but not AQ yield. We suspect that the enzymes involved in the biosynthesis of these two compounds responded differently to ultrasound treatment. We postulated that supplementation of the probable precursor CoQ0 along with ultrasound treatment during submerged fermentation should be able to to further stimulate the fungus to utilize CoQ0 to synthesize 4-AAQB. However, the results showed that ultrasound treatment with CoQ0 supplementation could not increase the yield of 4-AAQB. According to the results of LC-MS-MS analysis, CoQ0 was converted to CoQ3 and CoQ3B, therefore, it shuld be the precursor of both AQ and 4-AAQB. The fact that supplementation of CoQ0 could not increase 4-AAQB yield might be due to the accumulation of CoQ3B and AQB, because they could not further concvert to 4-AAQB. We could also deduce that the major influence of ultrasound treatment on polyketide pathway is to stimulate the intermediate 5-DMQ3B to convert to CoQ3B. Ultrasound treatment might also inhibit CoQ3 to convert to AQ. We also suspect that supplementation of CoQ0 along with ultrasound treatment would stimulate other metabolic pathways to utilize the CoQ0, which decreases the biosynthesis of CoQ3B, and thus decreases 4-AAQB yield. This combined treatment might also inhibit 5-DMQ3 and 6-DMQ3 to convert to CoQ3, and thus decreases AQ yield. | en |
dc.description.provenance | Made available in DSpace on 2021-06-15T11:36:52Z (GMT). No. of bitstreams: 1 U0001-1208202011085500.pdf: 3863686 bytes, checksum: 954b41be787ecfddbc311f7cd11aa5bc (MD5) Previous issue date: 2020 | en |
dc.description.tableofcontents | 目錄 謝誌 i 中文摘要 ii Abstract iii 第一章、前言 1 第二章、文獻整理 3 2.1 牛樟芝 3 2.1.1 牛樟芝簡介 3 2.1.2 牛樟芝液態發酵 3 2.1.3 牛樟芝液態發酵的影響因子 4 2.1.3.1 溫度 4 2.1.3.2 pH 值 5 2.1.3.3 攪拌速率與通氣量 6 2.1.3.4 碳源及氮源 7 2.2 牛樟芝的生理活性 7 2.2.1 抗氧化 8 2.2.2 抗發炎 8 2.2.3 免疫調節 9 2.2.4 肝臟保護 10 2.2.5 抗癌 11 2.3 牛樟芝的功效性成分 12 2.3.1 子實體功效性成分 12 2.3.2 菌絲體功效性成分 13 2.3.2.1 Antroquinonol 13 2.3.2.2 4-Acetylantroquinonol B 14 2.4 與 4-acetylantroquinonol B 及 antroquinonol 生合成相關之途徑 15 2.5 Coenzyme Q0 19 2.5.1 Coenzyme Q0簡介 19 2.5.2 Coenzyme Q0在牛樟芝菌絲體中扮演的角色 19 2.6 超音波 20 2.6.1 超音波簡介 20 2.6.2 超音波對生物體與發酵的影響 21 第三章、實驗構想及目的 24 3.1 實驗構想 24 3.2 實驗目的 25 3.3 實驗架構 26 第四章、材料與方法 28 4.1 實驗材料 28 4.1.1 牛樟芝菌株 28 4.1.2 實驗藥品 28 4.2 實驗方法 30 4.2.1 牛樟芝菌種保存 30 4.2.2 牛樟芝菌種活化 30 4.2.3 牛樟芝菌種繼代 31 4.2.4 牛樟芝液態發酵之菌酛製備 31 4.2.5 牛樟芝液態發酵 31 4.2.6 牛樟芝發酵期間之超音波處理 32 4.2.7 牛樟芝發酵液分析 32 4.2.7.1 pH 值測定 32 4.2.7.2 還原醣濃度測定 32 4.2.7.2.1 DNS 試劑配置 32 4.2.7.2.2 還原醣測定步驟 33 4.2.8 牛樟芝液態發酵菌絲體之分離與萃取 33 4.2.8.1 菌絲體與濾液分離 33 4.2.8.2 菌絲體生物質量測定 33 4.2.8.3 菌絲體乙醇萃取物製備 33 4.2.9 菌絲體乙醇萃取物 4-AAQB 及 AQ 之定量 34 4.2.9.1 HPLC 操作及分析條件 34 4.2.9.2 HPLC 分析之動相及流洗條件 34 4.2.10 菌絲體乙醇萃取物4-AAQB 及 AQ 生合成途徑中間產物分析 34 4.2.10.1 UPLC 分析條件 35 4.2.10.2 UPLC 管柱流洗條件 35 4.2.10.3 熱電噴灑游離器之參數 35 4.2.10.4 目標離子選擇之監控參數 35 4.2.10.5 平行反應監控參數 38 4.2.10.6 4-AAQB 及 AQ 生合成途徑中間產物資料 38 4.2.11 統計分析方法 39 第五章、結果與討論 40 5.1 超音波處理及添加 coenzyme Q0 對於液態發酵菌絲體之影響 40 5.1.1 發酵四週期間內發酵液 pH 值之變化 40 5.1.2 超音波處理及添加 coenzyme Q0 對於發酵液還原醣濃度之影響 42 5.1.3 超音波處理及添加 coenzyme Q0 對於菌絲體生物質量之影響 44 5.1.4 超音波處理及添加 coenzyme Q0 對於 4-AAQB 產率之影響 46 5.1.5 超音波處理及添加 coenzyme Q0 對於 AQ 產率之影響 48 5.2 添加不同濃度 coenzyme Q0 對牛樟芝液態發酵之影響 50 5.2.1 添加不同濃度 coenzyme Q0 對於發酵液 pH 值之影響 50 5.2.2 添加不同濃度 coenzyme Q0 對於發酵液還原醣濃度之影響 52 5.2.3 添加不同濃度 coenzyme Q0 對於菌絲體生物質量之影響 53 5.2.4 添加不同濃度 coenzyme Q0 對於 4-AAQB 產率之影響 55 5.2.5 添加不同濃度 coenzyme Q0 對於 AQ 產率之影響 56 5.2.6 小結 57 5.3 超音波處理並添加 coenzyme Q0 在不同發酵時間下對牛樟芝液態發酵之影響 58 5.3.1 發酵六週期間內發酵液 pH 值之變化 58 5.3.2 發酵六週期間內發酵液還原醣濃度之變化 60 5.3.3 在不同發酵時間下菌絲體生物質量之變化 61 5.3.4 在不同發酵時間下 4-AAQB 產率之變化 64 5.3.5 在不同發酵時間下 AQ 產率之變化 67 5.4 超音波處理及添加 coenzyme Q0 對牛樟芝 polyketide pathway 之影響 70 5.4.1 添加 coenzyme Q0 對 polyketide pathway 中間產物之影響 75 5.4.2 超音波處理對 polyketide pathway 中間產物之影響 76 5.4.3 超音波處理及添加 coenzyme Q0對 polyketide pathway中間產物之影響 76 第六章、結論 78 參考文獻 79 圖目錄 圖一、 AQ 與 4-AAQB 之結構 13 圖二、牛樟芝菌絲體中 4-AAQB 與 AQ 可能之生合成途徑 (Chou et al., 2019) 18 圖三、 Coenzyme Q0 之結構 19 圖四、超音波空蝕現象產生之原理 (Khan et al., 2018) 23 圖五、本實驗之主要概念 25 圖六、超音波處理及添加 coenzyme Q0 對發酵期間 pH 之影響 41 圖七、超音波處理及添加 coenzyme Q0 對於發酵第 28 天發酵液還原醣濃度之影響 43 圖八、超音波處理及添加 coenzyme Q0 對牛樟芝菌絲體生物質量之影響 45 圖九、超音波處理及添加 coenzyme Q0對牛樟芝菌絲體 4-AAQB 產率之影響 47 圖十、超音波處理及添加 coenzyme Q0對牛樟芝菌絲體 AQ 產率之影響 49 圖十一、添加 coenzyme Q0 對牛樟芝發酵液在 28 天發酵期間 pH 的影響 51 圖十二、添加 coenzyme Q0 對於發酵第 28 天發酵液還原醣濃度之影響 52 圖十三、添加 coenzyme Q0 對牛樟芝菌絲體生物質量之影響 54 圖十四、添加 coenzyme Q0對牛樟芝菌絲體 4-AAQB 產率之影響 55 圖十五、添加 coenzyme Q0對牛樟芝菌絲體 AQ 產率之影響 56 圖十六、牛樟芝發酵液在 6 週發酵期間 pH 的變化 59 圖十七、超音波處理及添加 coenzyme Q0 在不同時間點對還原醣濃度之影響 60 圖十八、超音波處理及添加 coenzyme Q0 在同一時間點對生物質量之影響 62 圖十九、每一組別在不同時間點下菌絲體生物質量之比較 63 圖二十、超音波處理及添加 coenzyme Q0 在同一時間點對 4-AAQB 產率之影響 65 圖二十一、每一組別在不同時間點下菌絲體 4-AAQB 產量之比較 66 圖二十二、超音波處理及添加 coenzyme Q0 在同一時間點對 AQ 產率之影響 68 圖二十三、每一組別在不同時間點下菌絲體 AQ 產量之比較 69 圖二十四、利用 LC-MS-MS 定量牛樟芝菌絲體中 4-AAQB 含量之結果 73 圖二十五、利用 LC-MS-MS 定量牛樟芝菌絲體中 AQ 含量之結果 74 表目錄 表一、HPLC 流洗條件 36 表二、UPLC 流洗條件 36 表三、質譜選擇離子監控參 37 表四、質譜選擇離子監控參數 37 表五、質譜平行反應監控參數 38 表六、牛樟芝菌絲體 polyketide pathway 中間產物 LC-MS-MS 分析資訊 (Chou et al., 2019) 38 表七、牛樟芝經不同處理後 polyketide pathway 中間產物含量與控制組之比較 72 表八、牛樟芝經不同處理後 polyketide pathway 上下游產物含量之比較 72 | |
dc.language.iso | zh-TW | |
dc.title | 探討牛樟芝菌絲體中 4-acetylantroquinonol B 生合成途徑 polyketide pathway 下游步驟之機制
| zh_TW |
dc.title | The Role of Downstream Steps of Polyketide Pathway in the Biosynthesis of 4-AAQB during Submerged Fermentation of Antrodia cinnamomea | en |
dc.type | Thesis | |
dc.date.schoolyear | 108-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 許輔(Fu Sheu),陳時欣(Shin-Hsin Chen),周繼中(Chi-Chung Chou),林育蔚(Yu-Wei Lin) | |
dc.subject.keyword | 牛樟芝,antroquinonol,4-acetylantroquinonol B,polyketide pathway,coenzyme Q0, | zh_TW |
dc.subject.keyword | Antrodia cinnamomea,antroquinonol,4-acetylantroquinonol B,polyketide pathway,coenzyme Q0, | en |
dc.relation.page | 86 | |
dc.identifier.doi | 10.6342/NTU202003060 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2020-08-17 | |
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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