請用此 Handle URI 來引用此文件:
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/69500
完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 呂廷璋 | |
dc.contributor.author | Hsin-Hua Lee | en |
dc.contributor.author | 李幸樺 | zh_TW |
dc.date.accessioned | 2021-06-17T03:17:29Z | - |
dc.date.available | 2023-07-06 | |
dc.date.copyright | 2018-07-06 | |
dc.date.issued | 2018 | |
dc.date.submitted | 2018-07-03 | |
dc.identifier.citation | 王鐘凰。菇蕈中多醣(1,3;1,6)-β-D-葡萄聚糖的分析與功能評估。國立臺灣大學博士論文。2014
李卓璟。以液相層析串聯質譜探討羅漢果皂素在酸與酵母菌模式中的結構轉換。國立台灣大學碩士論文。2009 林儀儂。培養條件對靈芝菌絲多醣與其(1,3;1,6)-β-D-聚葡萄醣生成量及性質之影響。國立台灣大學碩士論文。2004 林怡朱。靈芝不同生長階段液態培養菌絲與子實體之可溶性多醣特性。國立臺灣大學碩士論文。2005 林志彬。靈芝的現代研究,第三版,2007 邱群惠。靈芝或酵母菌對羅漢果皂苷之生物轉換及其應用。國立臺灣大學博士論文。2013 許俐菱。以豆科為基質之靈芝液態培養物之水溶性多醣特徵。國立臺灣大學碩士論文。2005 陳攸然。不同生長階段靈芝子實體水溶性多醣特性。國立臺灣大學碩士論文。2015 黃琬庭。結合酵素-層析法分析可食用菇類中具分支之(1,3;1,6)-β-D-葡萄聚糖含量。國立臺灣大學碩士論文。2013 黃美嫆。探討不同培養方法對靈芝菌絲和多醣體與靈芝酸產量之影響。中國藥學研究所碩士論文。2009 張毅偉。靈芝中具β-(1-6)分支之(1-3)-β-D-聚葡萄醣的性質與檢測。國立臺灣大學博士論文。2003 台灣質譜學會。質譜分析技術原理與應用,第一版,2015。 Babitskaya, V.; Shcherba, V.; Puchkova, T.; Smirnov, D., Polysaccharides of Ganoderma lucidum: factors affecting their production. Appl. Biochem. Biotechnol. 2005, 41 (2), 169-173. Bae, I. Y.; Kim, H. W.; Yoo, H. J.; Kim, E. S.; Lee, S.; Park, D. Y.; Lee, H. G., Correlation of branching structure of mushroom β-glucan with its physiological activities. Food Res. Intern. 2013, 51 (1), 195-200. Chen, Y.; Xie, M.-Y.; Gong, X.-F., Microwave-assisted extraction used for the isolation of total triterpenoid saponins from Ganoderma atrum. J. Food Eng. 2007, 81 (1), 162-170. Chen, Y.; Yan, Y.; Xie, M.-Y.; Nie, S.-P.; Liu, W.; Gong, X.-F.; Wang, Y.-X., Development of a chromatographic fingerprint for the chloroform extracts of Ganoderma lucidum by HPLC and LC–MS. J. Pharm. Biomed. Anal. 2008, 47 (3), 469-477. Cui, M.-l.; Yang, H.-y.; He, G.-q., Submerged fermentation production and characterization of intracellular triterpenoids from Ganoderma lucidum using HPLC-ESI-MS. Journal of Zhejiang University-SCIENCE B 2015, 16 (12), 998-1010. Cuong, T. D.; Hung, T. M.; Lee, J. H.; Woo, M. H.; Choi, J. S.; Kim, J.; Ryu, S. H.; Min, B. S., Inhibitory effect on NO production of triterpenes from the fruiting bodies of Ganoderma lucidum. Bioorg. Med. Chem. Lett. 2013, 23 (5), 1428-1432. Da, J.; Wu, W.-Y.; Hou, J.-J.; Long, H.-L.; Yao, S.; Yang, Z.; Cai, L.-Y.; Yang, M.; Jiang, B.-H.; Liu, X., Comparison of two officinal Chinese pharmacopoeia species of Ganoderma based on chemical research with multiple technologies and chemometrics analysis. J. Chromatogr. A 2012, 1222, 59-70. Dong, Q.; Wang, Y.; Shi, L.; Yao, J.; Li, J.; Ma, F.; Ding, K., A novel water-soluble β-d-glucan isolated from the spores of Ganoderma lucidum. Carbohydr. Res. 2012, 353, 100-105. Fang, Q. H.; Zhong, J. J., Two‐stage culture process for improved production of ganoderic acid by liquid fermentation of higher fungus Ganoderma lucidum. Biotechnol. Progr. 2002, 18 (1), 51-54. Fang, Q.-H.; Tang, Y.-J.; Zhong, J.-J., Significance of inoculation density control in production of polysaccharide and ganoderic acid by submerged culture of Ganoderma lucidum. Process Biochem. 2002, 37 (12), 1375-1379. Ferreira, I. C.; Heleno, S. A.; Reis, F. S.; Stojkovic, D.; Queiroz, M. J. R.; Vasconcelos, M. H.; Sokovic, M., Chemical features of Ganoderma polysaccharides with antioxidant, antitumor and antimicrobial activities. Phytochemistry 2015, 114, 38-55. Fraga, I.; Coutinho, J.; Bezerra, R. M.; Dias, A. A.; Marques, G.; Nunes, F. M., Influence of culture medium growth variables on Ganoderma lucidum exopolysaccharides structural features. Carbohydr. Polym. 2014, 111, 936-946. Gao, Q.; Araia, M.; Leck, C.; Emmer, Å., Characterization of exopolysaccharides in marine colloids by capillary electrophoresis with indirect UV detection. Anal. Chim. Acta 2010, 662 (2), 193-199. Gong, H.-G.; Zhong, J.-J., Hydrodynamic shear stress affects cell growth and metabolite production by medicinal mushroom Ganoderma lucidum. Chinese Chin. J. Chem. Eng. 2005, 13 (3), 426-428. Guo, X.-Y.; Han, J.; Ye, M.; Ma, X.-C.; Shen, X.; Xue, B.-B.; Che, Q.-M., Identification of major compounds in rat bile after oral administration of total triterpenoids of Ganoderma lucidum by high-performance liquid chromatography with electrospray ionization tandem mass spectrometry. J. Pharm. Biomed. Anal. 2012, 63, 29-39. Hirotani, M.; Furuya, T., Changes of the triterpenoid patterns during formation of the fruit body in Ganoderma lucidum. Phytochemistry 1990, 29 (12), 3767-3771. Isaka, M.; Chinthanom, P.; Kongthong, S.; Srichomthong, K.; Choeyklin, R., Lanostane triterpenes from cultures of the Basidiomycete Ganoderma orbiforme BCC 22324. Phytochemistry 2013, 87, 133-139. Jayasinghe, C.; Imtiaj, A.; Hur, H.; Lee, G. W.; Lee, T. S.; Lee, U. Y., Favorable culture conditions for mycelial growth of Korean wild strains in Ganoderma lucidum. Mycobiology 2008, 36 (1), 28-33. Jeong, Y.-T.; Jeong, S.-C.; Yang, B.-K.; Islam, R.; Song, C.-H., Optimal culture conditions for mycelial growth and exo-polymer production of Ganoderma applanatum. Mycobiology 2009, 37 (2), 89-93. Jo, W.-S.; Cho, Y.-J.; Cho, D.-H.; Park, S.-D.; Yoo, Y.-B.; Seok, S.-J., Culture conditions for the mycelial growth of Ganoderma applanatum. Mycobiology 2009, 37 (2), 94-102. Keypour, S.; Rafati, H.; Riahi, H.; Mirzajani, F.; Moradali, M. F., Qualitative analysis of ganoderic acids in Ganoderma lucidum from Iran and China by RP-HPLC and electrospray ionisation-mass spectrometry (ESI-MS). Food Chem. 2010, 119 (4), 1704-1708. Lee, K. M.; Lee, S. Y.; Lee, H. Y., Bistage control of pH for improving exopolysaccharide production from mycelia of Ganoderma lucidum in an air-lift fermentor. J. Biosci. Bioeng. 1999, 88 (6), 646-650. Lee, W. Y.; Park, Y.; Ahn, J. K.; Ka, K. H.; Park, S. Y., Factors influencing the production of endopolysaccharide and exopolysaccharide from Ganoderma applanatum. Enzyme Microb. Technol. 2007, 40 (2), 249-254. Leung, M.; Liu, C.; Koon, J.; Fung, K., Polysaccharide biological response modifiers. Immunol. Lett. 2006, 105 (2), 101-114. Li, C.-H.; Chen, P.-Y.; Chang, U.-M.; Kan, L.-S.; Fang, W.-H.; Tsai, K.-S.; Lin, S.-B., Ganoderic acid X, a lanostanoid triterpene, inhibits topoisomerases and induces apoptosis of cancer cells. Life Sci 2005, 77 (3), 252-265. Li, N.; Yan, C.; Hua, D.; Zhang, D., Isolation, purification, and structural characterization of a novel polysaccharide from Ganoderma capense. Int. J. Biol. Macromol. 2013, 57, 285-290. Liu, G.-Q.; Wang, X.-L.; Han, W.-J.; Lin, Q.-L., Improving the fermentation production of the individual key triterpene ganoderic acid Me by the medicinal fungus Ganoderma lucidum in submerged culture. Molecules 2012, 17 (11), 12575-12586. Liu, L.-Y.; Chen, H.; Liu, C.; Wang, H.-Q.; Kang, J.; Li, Y.; Chen, R.-Y., Triterpenoids of Ganoderma theaecolum and their hepatoprotective activities. Fitoterapia 2014, 98, 254-259. Liu, Y.; Liu, Y.; Qiu, F.; Di, X., Sensitive and selective liquid chromatography–tandem mass spectrometry method for the determination of five ganoderic acids in Ganoderma lucidum and its related species. J. Pharm. Biomed. Anal. 2011, 54 (4), 717-721. Maruyama, H.; Yamazaki, K.; Murofushi, S.; Konda, C.; Ikekawa, T., Antitumor activity of Sarcodon aspratus (BERK.) S. ITO and Ganoderma lucidum (FR.) KARST. J. Pharmacobio-Dyn. 1989, 12 (2), 118-123. Mdachi, S. J.; Nkunya, M. H.; Nyigo, V. A.; Urasa, I. T., Amino acid composition of some Tanzanian wild mushrooms. Food Chem. 2004, 86 (2), 179-182. Papaspyridi, L. M.; Katapodis, P.; Gonou‐Zagou, Z.; Kapsanaki‐Gotsi, E.; Christakopoulos, P., Growth and biomass production with enhanced β‐glucan and dietary fibre contents of Ganoderma australe ATHUM 4345 in a batch‐stirred tank bioreactor. Eng. Life Sci. 2011, 11 (1), 65-74. Peng, L.; Qiao, S.; Xu, Z.; Guan, F.; Ding, Z.; Gu, Z.; Zhang, L.; Shi, G., Effects of culture conditions on monosaccharide composition of Ganoderma lucidum exopolysaccharide and on activities of related enzymes. Carbohydr. Polym. 2015, 133, 104-109. Raja, G. A.; Aravindan, R.; Viruthagiri, T., Batch and continuous production of Exopolysaccharide by Ganoderma lucidum MTCC 1091 in submerged fermentation. Indian J. Pharm. Educ. Res. 2007, 41 (3), 211-218. Su, H.-J.; Fann, Y.-F.; Chung, M.-I.; Won, S.-J.; Lin, C.-N., New Lanostanoids of Ganoderma tsugae. J. Nat. Prod. 2000, 63 (4), 514-516. Tang, Y. J.; Zhong, J. J., Scale‐Up of a Liquid Static Culture Process for Hyperproduction of Ganoderic Acid by the Medicinal Mushroom Ganoderma lucidum. Biotechnol. Progr. 2003, 19 (6), 1842-1846. Tang, Y.-J.; Zhang, W.; Liu, R.-S.; Zhu, L.-W.; Zhong, J.-J., Scale-up study on the fed-batch fermentation of Ganoderma lucidum for the hyperproduction of ganoderic acid and Ganoderma polysaccharides. Process Biochem. 2011, 46 (1), 404-408. Tang, Y.-J.; Zhong, J.-J., Fed-batch fermentation of Ganoderma lucidum for hyperproduction of polysaccharide and ganoderic acid. Enzyme Microb. Technol. 2002, 31 (1-2), 20-28. Taylor, P. R.; Tsoni, S. V.; Willment, J. A.; Dennehy, K. M.; Rosas, M.; Findon, H.; Haynes, K.; Steele, C.; Botto, M.; Gordon, S., Dectin-1 is required for β-glucan recognition and control of fungal infection. Nat. Immunol. 2007, 8 (1), 31. Volman, J. J.; Ramakers, J. D.; Plat, J., Dietary modulation of immune function by β-glucans. Physiol. Behav. 2008, 94 (2), 276-284. Wang, C.-L.; Lu, C.-Y.; Hsueh, Y.-C.; Liu, W.-H.; Chen, C.-J., Activation of antitumor immune responses by Ganoderma formosanum polysaccharides in tumor-bearing mice. Appl. Microbiol. Biotechnol. 2014, 98 (22), 9389-9398. Wang, Y.-Y.; Khoo, K.-H.; Chen, S.-T.; Lin, C.-C.; Wong, C.-H.; Lin, C.-H., Studies on the immuno-modulating and antitumor activities of Ganoderma lucidum (Reishi) polysaccharides: functional and proteomic analyses of a fucose-containing glycoprotein fraction responsible for the activities. Bioorg. Med. Chem. 2002, 10 (4), 1057-1062. Wei, Z.-h.; Duan, Y.-y.; Qian, Y.-q.; Guo, X.-f.; Li, Y.-j.; Jin, S.-h.; Zhou, Z.-X.; Shan, S.-y.; Wang, C.-r.; Chen, X.-j., Screening of Ganoderma strains with high polysaccharides and ganoderic acid contents and optimization of the fermentation medium by statistical methods. Bioprocess Biosyst. Eng. 2014, 37 (9), 1789-1797. Wei, Z.-h.; Duan, Y.-y.; Qian, Y.-q.; Guo, X.-f.; Li, Y.-j.; Jin, S.-h.; Zhou, Z.-X.; Shan, S.-y.; Wang, C.-r.; Chen, X.-j., Screening of Ganoderma strains with high polysaccharides and ganoderic acid contents and optimization of the fermentation medium by statistical methods. Bioprocess Biosyst. Eng. 2014, 37 (9), 1789-1797. Xu, J.-W.; Xu, Y.-N.; Zhong, J.-J., Production of individual ganoderic acids and expression of biosynthetic genes in liquid static and shaking cultures of Ganoderma lucidum. Appl. Microbiol. Biotechnol. 2010, 85 (4), 941-948. Xu, J.-W.; Zhao, W.; Zhong, J.-J., Biotechnological production and application of ganoderic acids. Appl. Microbiol. Biotechnol. 2010, 87 (2), 457-466. Yang, F.-C.; Liau, C.-B., The influence of environmental conditions on polysaccharide formation by Ganoderma lucidum in submerged cultures. Process Biochem. 1998, 33 (5), 547-553. Yang, M.; Wang, X.; Guan, S.; Xia, J.; Sun, J.; Guo, H.; Guo, D.-a., Analysis of triterpenoids in Ganoderma lucidum using liquid chromatography coupled with electrospray ionization mass spectrometry. J. Am. Soc. Mass Spectrom. 2007, 18 (5), 927-939. Yuan, B.; Chi, X.; Zhang, R., Optimization of exopolysaccharides production from a novel strain of Ganoderma lucidum CAU5501 in submerged culture. Braz. J. Microbiol. 2012, 43 (2), 490-497. Zhao, W.; Xu, J.-W.; Zhong, J.-J., Enhanced production of ganoderic acids in static liquid culture of Ganoderma lucidum under nitrogen-limiting conditions. Bioresour. Technol. 2011, 102 (17), 8185-8190. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/69500 | - |
dc.description.abstract | 靈芝為中國傳統食用藥材,紫芝(Ganoderma formosanum)為臺灣特有種,而赤芝(Ganoderma lucidium)為常見之藥材及保健食品,被廣為研究及有效成分為靈芝酸以及多醣。本研究目的為建立一套靈芝酸分析平台,使用液相層析串聯質譜(HPLC-MS/MS)分析靈芝酸的種類,在特定能量下結因構不同使得斷片模式有所差異;另外使用高解析質譜(HRMS)得精確分子量以及紫外光吸收來推測靈芝酸種類以及含量。另一個具有生理活性成分為多醣體,其中廣泛被認為具有調節免疫結構之多醣體為(1,3;1,6)-β-D-葡聚醣分支度介於0.2-0.33,本實驗使用專一性酵素水解(1,3;1,6)-β-D-葡聚醣可精確得知含量並得知分支度。子實體生長時程曠日費時,但是液態醱酵具有培養時間快,分離萃取容易等優點。許多文獻研究指出靜置培養靈芝使得靈芝酸的產量增加,因此本實驗使用靜置培養方式產生靈芝酸及(1,3;1,6)-β-D-葡聚醣。搖瓶培養下菌絲體與多醣體生產較為快速以及大量,兩階段培養方式有利生產目標產物。本實驗研究目的在比較紫芝以及赤芝在三種不同培養方式下,菌絲體生成量、菌絲體中多醣體、醱酵液中多醣體、(1,3;1,6)-β-D-葡聚醣含量及其分支度以及靈芝酸含量與其種類的差異。結果顯示紫芝在靜置培養下菌絲體最多,並且第十天下菌絲體及醱酵液中多醣體最多,赤芝則是在搖瓶培養第十五天下菌絲體最多,並且菌絲體多醣體在第十天最多,但是醱酵液並沒有顯著差異。紫芝產(1,3;1,6)-β-D-葡聚醣,在靜置培養下第三十天含量最高並且顯著高於赤芝,並且分支度落於0.2-0.33之間,但是赤芝只有在靜置培養下才有(1,3;1,6)-β-D-葡聚醣產生並且分支度並低於0.2-0.33。整體而言赤芝菌絲體含量以及多醣含量高於紫芝,但是紫芝所產多醣體中(1,3;1,6)-β-D-葡聚醣佔多醣體比例高於赤芝。使用超高效能液相串聯質譜儀(UHPLC-MS/MS)靈芝酸分析方法平台中,從松杉靈芝以及赤芝子實體使用酒精萃取靈芝酸液中建立品管樣品,品管樣品中推測可能含有靈芝酸27種,並且使用標準添加法測品管樣品中,具有靈芝酸標準品種類的真實濃度以及相對定量無標準品之靈芝酸,另外回收率推測質譜訊號影響樣品基質的結果收吸波峰的特徵篩選波峰,進一步利用高解析質譜(HRMS)分析其精確分子量,此方法理論分子質荷比(m/z)與實際測試m/z的偏差值在-2.33~ 2.30 ppm之間。最後在本實驗培養方法下,兩株菌在三種培養方式下以紫芝靜置培養30天生成Ganoderi acid H 0.99 ppm最多,並且高於赤芝搖瓶培養的0.23 ppm,赤芝靜置培養以及搖瓶培養下生成Ganoderic acid H 則無顯著差異。 | zh_TW |
dc.description.abstract | Ganoderma formosanum and Ganoderma lucidium, Lingzhi, are traditional Chinese medicine and are considered as healthy food. The bioactive components in Lingzhi are (1, 3;1, 6)-β-D-glucans and ganoderic acids. This study aims to investigate that the effect of the static culture, the submerged culture and the two-stage fermentation on the production of the (1, 3;1, 6)-β-D-glucans and ganoderic acids of Ganoderma formosanum and Ganoderma lucidum. The biomass production, polysaccharide in mycelium and broth, the content and the degree of the branch of (1,3;1,6)-β-D-glucans and the structure of ganoderic acids of Ganoderma formosanum and Ganoderma lucidum were compared. In this study, the (1,3)-β-D-glucanase was used to investigate the content and the degree of the branch of (1,3;1,6)-β-D-glucans. Liquid chromatography-tandem mass spectrometry was used to analyze ganoderic acids. The structure properties of ganoderic acids were identified by accurate m/z (-2.33-2.33 ppm), MS/MS spectra and UV spectra. Standard addition method was used to analyze the concentration of ganoderic acids. Relative quantification method was used to analyze the ganoderic acids without an available commercial standard. In this research, the result shows that static culture produces more ganoderic acid than submerge culture. However, shaking culture produces more biomass and polysaccharide. Two-stage fermentation can enhance the yield of the target compound. The mycelium of Ganoderma formosanum has maximum biomass by static culture. At 10 days, Ganoderma formosanum has the maximum yield of polysaccharides in the mycelium and the broth. By shaking culture, the maximum biomass of the mycelium of Ganoderma lucidum is at 15 days and the maximum yield of the polysaccharide of Ganoderma lucidum is at 10 days but there is no significant difference in the broth. At 30 days, by static culture, the content of (1,3;1,6)-β-D-glucan of Ganoderma formosanum mycelia is more than Ganoderma lucidum. The degree of the branch of (1,3;1,6)-β-D-glucan of Ganoderma formosanum mycelia is between 0.2-0.33. Ganoderma lucidum only produces (1,3;1,6)-β-D-glucan in static culture and the degree of the branch of (1,3;1,6)-β-D-glucan is lower than 0.2-0.33. Ganoderma lucidum contains more mycelia and polysaccharides than Ganoderma formosanum. However, the percentage of (1,3;1,6)-β-D-glucan in the polysaccharides of Ganoderma formosanum is more than Ganoderma lucidum. Ganoderma lucidum and Ganoderma tsugae fruiting body were used as QC sample. 27 kinds of ganoderic acids were found in QC sample. In this study, the mycelium of Ganoderma formosanum in static culture 30 days has more ganoderic acid H than Ganoderma lucidum. The static culture and shaking culture have no significant effect on the production of Ganoderic acid H of Ganoderma lucidum. | en |
dc.description.provenance | Made available in DSpace on 2021-06-17T03:17:29Z (GMT). No. of bitstreams: 1 ntu-107-R03641029-1.pdf: 3388445 bytes, checksum: 05ef3e393eb4c39e91ab4a3bf10c0559 (MD5) Previous issue date: 2018 | en |
dc.description.tableofcontents | 摘要 i
Abstract iii 目錄 I 圖目錄 XIII 表目錄 XIX 一. 前言 1 二. 文獻回顧 2 2.1. 靈芝介紹 2 2.2. 生物分類及構造 3 2.3. 靈芝生長 3 2.3.1野生靈芝 3 2.3.2人工靈芝 4 2.4. 生物活性成分與功能 4 2.4.1多醣體 4 2.4.2三萜類 5 2.4.2.1結構與斷片模式 6 2.4.3分析方法 10 2.4.4質譜儀及串聯質譜 11 2.4.5高解析質譜 15 2.4.6三萜類生理活性 18 2.5. 靈芝液態培養因子 19 2.5.1溫度 19 2.5.2酸鹼值 19 2.5.3碳源 20 2.5.4氮源 20 2.5.5碳氮比 21 2.5.6醱酵方式 21 2.5.6.1連續式醱酵 21 2.5.6.2饋料式醱酵 22 2.5.6.3批次醱酵 22 2.5.6.4靜置培養 22 2.5.7接菌量 24 2.5.8剪切力 24 2.5.9通氣量 25 2.5.10兩階段醱酵 25 2.6. (1,3)-β-D-葡聚醣 28 2.6.1(1,3)-β-D-葡聚醣結構與活性 29 2.6.1.1分子量 29 2.6.1.2分支度 29 2.6.1.3構型 30 2.6.2免疫機制 31 2.6.3(1,3)-β-D-葡聚醣分析 32 2.6.3.1螢光測定法(aniline blue) 32 2.6.3.2Congo red呈色法 32 2.6.3.3酵素串聯高效液相層析 33 2.6.4真菌中(1,3;1,6)-β-D-葡聚醣 34 2.7. 巨噬細胞其免疫反應 45 三. 研究目的與實驗架構 46 3.1研究目的 46 3.2實驗架構 46 3.3研究假設 47 3.3.1第一階段:紫芝及赤芝在培養方式下生長曲線 47 3.3.2第二階段:多醣及(1,3;1,6)-β-D-葡聚醣含量及分支度分析 47 3.3.3第三階段:靈芝酸分析方法建立及樣品分析 47 四. 第一部份:紫芝與赤芝使用不同培養方式對於菌絲生成及(1,3;1,6)-β-D-葡聚醣含量及分支度結果 48 4.1材料與方法 48 4.1.1靈芝 48 4.1.1.1菌種來源 48 4.1.1.2菌絲體培養 48 4.1.2試藥與試劑 48 4.1.3標準品及酵素 49 4.1.4儀器 49 4.1.4.1高效能陰離子交換層析儀-脈衝安培檢測法 49 4.1.5靈芝菌絲體醱酵培養 50 4.1.5.1液態醱酵培養基 50 4.1.6醱酵菌絲體樣品製備 50 4.1.6.1菌絲體活化 50 4.1.6.2搖瓶培養醱酵(Shaking culture; SK) 50 4.1.6.3靜置培養醱酵(Static culture; ST) 50 4.1.6.4兩階段式醱酵(Two-stage culture; TS) 50 4.1.7分析方法 52 4.1.7.1菌絲體均質及醱酵液儲存方式 52 4.1.7.2菌絲體重量 52 4.1.7.3殘醣含量測定 52 4.1.7.4總醣含量測定 52 4.1.7.5(1,3;1,6)-β-D-聚葡萄醣含量與分支度-酵素層析法 53 4.2結果與討論 55 4.2.1菌絲體生成量 55 4.2.2菌絲體及醱酵液中多醣含量比較 57 4.2.3不同靈芝在相同培養方式下菌絲體及多醣結果 62 4.2.4菌絲體中 (1,3;1,6)-β-D-葡萄聚醣結果 70 五. 第二部份:靈芝酸分析方法平台建立及樣品分析 77 5.1材料與方法 77 5.1.1溶劑 77 5.1.2標準品 77 5.1.3儀器 77 5.1.3.1超高效液相層析分析串聯質譜(UHPLC-MS) 77 5.1.3.2超高液相層析分析串聯高解析質譜儀(UHPLC-HRMS) 77 5.1.4品管樣品 78 5.1.5超高效能液相層析及質譜儀設定(UPLC-MS/MS) 79 5.1.5.1液相層析 79 5.1.5.2檢測器方法設定 79 5.1.5.3檢測器及高解析質譜譜方法設定 81 5.1.5.4靈芝酸軟體比對設定 82 5.1.6靈芝酸分析條件方法 82 5.1.6.1層析條件測試 82 5.1.7分析平台確校實驗方法 83 5.1.7.1標準曲線 83 5.1.7.2偵測極限及定量極限 83 5.1.7.3標準添加法 83 5.1.7.4回收率 84 5.2結果與討論 85 5.2.1靈芝酸層析條件測試結果 85 5.2.2機台的標準曲線 88 5.2.3標準添加法 90 5.2.4偵測極限及定量極限 90 5.2.5回收率 92 5.2.6品管樣品中靈芝酸結果 95 5.2.6.1第一類:代表靈芝酸ganoderic acid A & ganoderic acid C2 96 5.2.6.2第二類:代表靈芝酸ganoderic acid AM1& ganoderic acid J 98 5.2.6.3第三類:代表靈芝酸ganoderic acid B, D,G& K 99 5.2.6.4第四類:代表靈芝酸 Ganoderic acid H 101 5.2.6.5第五類:代表靈芝酸lucidenic acid N 103 5.2.6.6第六類:其他 105 5.2.7高解析質譜結果 114 5.2.7.1標準曲線 115 5.2.7.2品管樣品精確分子量結果 116 5.2.8醱酵樣品中靈芝酸結果 118 六. 結論 121 七. 參考文獻 123 八. 附錄 131 8.1高解析質譜分析方法設定 131 8.2品管樣品中靈芝酸結果(按照分子量排) 137 8.2.1 Lucidenic acid F 137 8.2.2. Lucidenic acid A 139 8.2.3. Lucidenic acid N 141 8.2.4.7,15-Dihydroxy-4,4,14-teimethyl-3,11-dioxochol-8-en-24-oic acid 143 8.2.5. Ganolucidenic acid A 145 8.2.6. Ganolucidic acid D 147 8.2.7. Ganoderenic acid D 149 8.2.8. Ganoderenic acid B 151 8.2.9. Ganoderic acid AM1 153 8.2.10. Ganoderic acid D 155 8.2.11. Lucidenic acid D 157 8.2.12. Ganoderic acid J 159 8.2.13. Lucidenic acid E 161 8.2.14. Ganoderic acid B 163 8.2.15. Ganoderic acid A 165 8.2.16. Ganoderic acid C2 167 8.2.17. Elfvingic acid A 169 8.2.18. 12-Hdroxy-3,7,11,15,23-pentaoxo-lanost-8-en-26-oic acid 171 8.2.19. Ganoderic acid C6 173 8.2.20. 12-Hydroxyganoderic acid D 175 8.2.21. Ganoderic acid G 177 8.2.22. 12-Hydroxyganoderic acid C2 179 8.2.23. Ganoderic acid K 181 8.2.24. Ganoderic acid F 183 8.2.25. Ganoderic acid H 185 8.2.26. Ganoderenic acid K 187 8.2.27. 3-Acethlganoderic acid H 189 | |
dc.language.iso | zh-TW | |
dc.title | 培養條件對紫芝與赤芝菌絲生成多醣與靈芝酸含量與組成的影響 | zh_TW |
dc.title | Effects of culture conditions on production and composition of polysaccharides and ganoderic acids of Ganoderma formosanum and Ganoderma lucidum | en |
dc.type | Thesis | |
dc.date.schoolyear | 106-2 | |
dc.description.degree | 碩士 | |
dc.contributor.coadvisor | 鄭光成 | |
dc.contributor.oralexamcommittee | 潘敏雄,羅翊禎,王惠珠 | |
dc.subject.keyword | Ganoderma formosanum,Ganoderma lucidium,LCMS/MS,靈芝酸,(1,3,1,6)-β-D-葡聚醣,分支度,多醣, | zh_TW |
dc.subject.keyword | Ganoderma formosanum,Ganoderma lucidium,LCMS/MS,ganoderic acid,(1,3,1,6)-β-D-glucan,DB (degree of branch),polysaccharides., | en |
dc.relation.page | 190 | |
dc.identifier.doi | 10.6342/NTU201800977 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2018-07-03 | |
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 |
文件中的檔案:
檔案 | 大小 | 格式 | |
---|---|---|---|
ntu-107-1.pdf 目前未授權公開取用 | 3.31 MB | Adobe PDF |
系統中的文件,除了特別指名其著作權條款之外,均受到著作權保護,並且保留所有的權利。