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研究生:潘厚良
研究生(外文):PAN,HOU-LIANG
論文名稱:環境與代謝控制策略對烏靈參液態發酵菌絲體與生物活性成分之影響
論文名稱(外文):Effects of Environmental and Metabolism Control Strategy on Mycelial Biomass and Bioactive Ingredients Production by the Submerged Culture of Xylaria nigripes
指導教授:徐泰浩徐泰浩引用關係林芳儀林芳儀引用關係
指導教授(外文):XU,TAI-HAOLIN,FANG-YI
口試委員:徐泰浩林芳儀陳勁初李世傑
口試委員(外文):XU,TAI-HAOLIN,FANG-YIZHEN,JIN-CHULI,SHI-JIE
口試日期:2014-07-24
學位類別:碩士
校院名稱:大葉大學
系所名稱:生物產業科技學系
學門:生命科學學門
學類:生物科技學類
論文種類:學術論文
論文出版年:2014
畢業學年度:102
語文別:中文
論文頁數:92
中文關鍵詞:烏靈參液態發酵菌絲體恆定pH溫度生物活性成分
外文關鍵詞:Xylaria nigripessubmerged fermentatioBiomasspH-stattemperaturebiological
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  烏靈參(Xylaria nigripes) 又名黑柄炭角菌,其分類屬子囊菌亞門(Ascomycotina)、炭角菌科(Xylariaceae)、炭角菌屬(Xylaria),是一種生長在地下深處、土棲白蟻廢棄蟻巢上的珍貴藥用真菌。依過去中藥典籍記載,烏靈參具有鎮靜安神、補氣固腎、養血、健脾除濕等功效,可治療憂鬱、神經衰弱、失眠及心悸等症。
  烏靈參因生長環境特殊且資源十分缺乏,相關文獻稀少,液態發酵烏靈參文獻又更加稀少,因此本實驗目的主要探討,烏靈參於液態發酵進行環境因子試驗並且放大產程於20L發酵槽進而分析菌絲體和生物活性物生成量之影響。
在溫度調控、不同pH、雙液相選擇與濃度、接菌量、通氣量和培養時間各試驗因子,實驗結果顯示烏靈參液態發酵於20L發酵槽中,在條件於溫度25℃漸進減少、恆定pH為4、添加0.1%大豆油、接菌量10%、通氣量2vvm、培養5天,其生物質量、胞外多醣、胞內多醣、總多酚、粗三萜、腺苷含量分別為2.43mg/mL、1.75mg/mL、46.42mg/g、4.81mg/g、37.36mg/g、4.31mg/g。
  由實驗結果顯示,最適培養條件與初始培養條件培養烏靈參相比之下,除了胞外多醣外,菌絲量與其餘生物活性成分皆可提升將近4倍以上生成量。

  Xylaria nigripes, named Wū-Línɡ-Shēn in Chinese, belongs to the division of Ascomycotina, the family of Xylariaceae, and down to genus of Xylaria. X. nigripes is a precious medicinal fungus that grows on top of the abandoned termite nests underground. Based on the Chinese pharmacopeia, Wū-Línɡ-Shēn has several biological functions such as tranquilizer, enhancing physical energy, maintaining renal function, dehumidification effect in spleen, treating depression, neurasthenia, and insomnia and palpitations embolism.
  Due to its difficulty to plant and less productive quantity, there is a little of information about Wū-Línɡ-Shēn. Researchers studying on liquid state-fermented of Wū-Línɡ-Shēn is few. The major purpose of this study is to test the environmental factors for Wū-Línɡ-Shēn to culture under liquid state fermentation. Then, the production was scaled up to a 20L fermenter and further analyzed the contents of mycelium biomass and bioactive ingredients.
  The experiments were conducted based on factors, including temperature, pH, two-phase selectivity and concentration, volume of inoculation, ventilation, and cultivation time. The results indicated the optimized condition for optimal production were 5 day’s growing, temperature at 25 degree Celsius, pH-state at 4, and 2vvm ventilation with 0.1% soybean oil, 10% inoculation. The amount of biomass, extracellular polysaccharide, intracellular polysaccharide, total polyphenol, crude triterpenoid, and adenosine was 2.43mg/mL、1.75mg/mL、46.42mg/g、4.81mg/g、37.36mg/g and 4.319mg/g, respectively.
  In conclusion, compared to standardized procedure, we except the extracellular polysaccharide, the yield of mycelium and other bioactive ingredients would increase up to four times at liquid state fermentation of Wū-Línɡ-Shēn in 20L fermenter.

封面內頁
簽名頁
中文摘要iii
英文摘要iv
誌謝v
目錄vi
圖目錄x
表目錄xiii

1.前言1
2.文獻回顧2
2.1烏靈參簡介2
2.1.1烏靈參分類2
2.1.2形態特徵2
2.2烏靈參的培養4
2.2.1液態培養4
2.2.2發酵工程5
2.3烏靈參液態發酵之環境因子6
2.3.1溫度6
2.3.2 pH值6
2.3.3雙液相7
2.3.4接菌量7
2.3.5通氣量8
3.材料與方法9
3.1 實驗架構9
3.2 實驗材料10
3.2.1 實驗菌株10
3.2.2 實驗藥品10
3.2.3儀器設備11
3.3培養基組成13
3.4菌株培養14
3.4.1烏靈參菌株培養14
3.5基礎培養條件14
3.5.1搖瓶基礎培養條件14
3.5.2 20L發酵槽基礎培養條件15
3.6搖瓶培養試驗15
3.6.1不同溫度對於烏靈參液態發酵菌絲體、胞外多醣、最終pH值之影響15
3.6.2不同初始pH對於烏靈參液態發酵菌絲體、胞外多醣、最終pH值之影響15
3.6.3雙液相發酵對於烏靈參液態發酵菌絲體與生物活性成分之影響16
3.6.4不同接菌量對於烏靈參液態發酵菌絲體與生物活性成分之影響16
3.6.5不同接菌量與培養時間對於烏靈參液態發酵菌絲體與生物活性成分之影響16
3.7 20L發酵槽培養試驗17
3.7.1不同饋菌量與培養時間對於烏靈參液態發酵菌絲體與生物活性成分之影響17
3.7.2添加不同濃度的沙拉油對於烏靈參液態發酵菌絲體與生物活性成分之影響17
3.7.3不同進氣量對於烏靈參液態發酵菌絲體與生物活性成分之影響18
3.7.4不同恆定pH對於烏靈參液態發酵菌絲體與生物活性成分之影響18
3.7.5恆定pH與漸進調控溫度對於烏靈參液態發酵菌絲體與生物活性成分之影響18
3.7.6恆定pH與瞬間調控溫度對於烏靈參液態發酵菌絲體與生物活性成分之影響19
3.8 分析方法19
3.8.1 pH值之測定19
3.8.2 菌絲體生物質量測定與製備19
3.8.3 胞內、外多醣含量測定與樣品製備20
3.8.3.4 胞外多醣之澱析21
3.8.4 總多酚含量測定與樣品製備21
3.8.5 粗三萜含量測定與樣品製備22
3.8.6 腺苷含量測定與樣品製備23
4.結果與討論25
4.1烏靈參液態發酵之搖瓶試驗25
4.1.1不同溫度對於烏靈參液態發酵菌絲體、胞外多醣、最終pH值之探討25
4.1.2不同初始pH對於烏靈參液態發酵菌絲體、胞外多醣、最終pH值之探討29
4.1.3雙液相發酵對於烏靈參液態發酵菌絲體與生物活性物之探討33
4.1.4不同接菌量對於烏靈參液態發酵菌絲體與生物活性物之探討38
4.1.5不同接菌量與培養時間對於烏靈參液態發酵菌絲體與生物活性物之探討43
4.2烏靈參液態發酵之二十公升發酵槽試驗48
4.2.1不同接菌量與培養時間對於烏靈參液態發酵菌絲體與生物活性物之探討48
4.2.2添加不同濃度大豆油對於烏靈參液態發酵菌絲體與生物活性物之探討52
4.2.3不同通氣量對於烏靈參液態發酵菌絲體與生物活性物之探討57
4.2.4不同恆定pH對於烏靈參液態發酵菌絲體與生物活性物之探討62
4.2.5恆定pH與調控溫度67
4.2.6初始條件與實驗結果對於烏靈參液態發酵菌絲體與生物活性成分之影響81
5.結論87
參考文獻88



1.王培銘。2002。食藥用菇液態培養製程之開發。食品工業34:31-35。
2.尹軍華。2008。黑柄炭角菌菌絲體水溶性多糖的分離純化與結構鑑定。湖南師範大學碩士學位論文。中國湖南。
3.朱志熊、張澤文、張平。2005。黑柄炭角菌的菌種分離及其培養特性。中國食用菌24:15-18。
4.李衛旗、吳學謙、馮依力、程俊文、賀亮。2010。靈芝胞外多糖雙液相發酵體系的特性。浙江大學學報(理學版)6:686-690
5.何育澤。2012。豆類與穀物基質應用於樟芝與銀耳共培養液態發酵與其生物活性探討。大葉大學碩士論文 。台灣彰化。
6.林嘉隆。2005。培養條件對雲芝菌絲體生長與多醣生成之影響。東海大學碩士論文。台灣台中。
7.林怡朱。2005。靈芝不同生長階段液態培養菌絲與子實體之水溶性多醣特性。國立台灣大學碩士論文。台灣台北。
8.周少奇、馮樸蓀、唐立民。1997。靈芝菌絲體液體懸浮培養發酵液的流變特性。高校化學工程學報。11:197-200。
9.翁榕安。2009。黑柄炭角菌的人工栽培技術及部分活性成分的研究。湖南師範大學碩士論文。中國湖南。
10.陳宛如、李振唐、史美中、倪建勇。1988。真菌中藥鳥靈參的生物學特性研究。現代應用醫學5:10-13。
11.馬橙。2011。黑炳炭角菌生活史研究。湖南師範大學碩士論文。中國湖南。
12.馬志章、左萍萍、陳宛如。1999。烏靈菌粉的鎮靜作用及其機理研究.中國藥學雜誌 34(6):374-377。
13.陳怡倩。2001。利用批式液態培養來探討檸檬酸對裂褶菌生長及其多醣體生成影響之研究。國立中央大學碩士論文。台灣桃園。
14.陳書豪。2006。探討樟芝的溫度變化對液態發酵與固態發酵生產三萜類與多醣體之影響。國立中央大學碩士論文。台灣桃園。
15.陸震鳴。2009。樟芝深層液態培養及其三萜類化合物的研究。江南大學論文博士。大陸江南。
16.張澤文。2006。黑柄炭角菌生物學特性及其活性物質的初步研究。湖南師範大學碩士論文。大陸湖南。
17.許淳均。2004。探討培養基組成對巴西洋菇發酵生產活性多醣及其特性之影響。國立中央大學博士論文。台灣桃園。
18.黃思齊。2011。發酵產程擴大及不同培養基對雲芝胞外醣肽化學特性之影響。大葉大學碩士論文。台灣彰化。
19.黃麗娜。1998。菇類菌絲體深層發酵在食品工業上之應用。食藥用菇類培養與應用144-150。
20.鄔家林。1984。真菌中藥—烏靈參。成都中醫學院學報33-35。
21.楊雅萍。2008。烏笭參之最佳液態發酵條件研究。大仁科技大學碩士論文。台灣屏東。
22.楊士賢。2006。影響雲芝菌絲已與多醣批次饋料發酵程控制參數因子之探討。大葉大學碩士論文。台灣彰化。
23.劉忠義、曾虹燕。2002。茯苓液體培養研究初探。湘潭大學食品科學與工程系,湖南湘潭。
24.Bao, H.N., Ochiai, Y. and Ohshima, T. 2010. Antioxidative activities of hydrophilic extracts prepared from the fruiting body and spent culture medium of Flammulina velutipes. Bioresour. Technol. 101:6248–6255.
25.Chain, E. B.,Gualandi, G. and Morisi, G.1966. Aeration studies. IV.Aeration condition in 3000-liter submerged fermentation with various microorganisms. Biotechnology and Bioengineering,595-619.
26.Chienyan Hsieh , Hui-Liang Wang , Chien-Cheng Chen, Tai-Hao Hsu and Mei-Hua Tseng . 2008.Effect of plant oil and surfactant on the production of mycelial biomass and polysaccharides in submerged culture of Grifola frondosa.Biochemical Engineering Journal. 38: 198–205.
27.Chienyan Hsieh , Chia-Jang Liu, Mei-Hua Tseng, Chaur-Tsuen Lo and Yuan-Chang Yang.2006. Effect of olive oil on the production of mycelial biomass and polysaccharides of Grifola frondosa under high oxygen concentration aeration. Enzyme and Microbial Technology. 39: 434–439.
28.Chen NY, Hsu TH, Lin FY, Lai H-H and Wu JY. 2006. Effects on cytokine-stimulating activities of EPS from Tremella mesenterica with various carbon sources.99:92-97
29.Dubois M, Gilles KA, Hamilton JK, Rebers, PA and Smith, F. 1956.Colorimetric method for determination of sugars and related substances. Analytical Chemistry.28:350-356.
30.Fang, Q. H. and Zhong, J. J.2002. Effect of initial pH on production of ganoderic acid and polysaccharide by submerged fermentation of Ganoderma lucidum. Bioprocess Biochemistry. 37:769-774.
31.Fan, JP and He, CH. 2006. Simultaneous quantification of three major bioactive triterpene acids in the leaves of Diospyros kaki by high-performance liquid chromaography method.41:950-956.
32.Glazebrook, M.A., Vining, L.C. and White, R.L. 1992. Growth morphology of Streptomyces akiyoshiensis in submerged culture: influence of pH, inoculum, and nutrients. Canadian journal of microbiology. 38: 98-103.
33.Hu Zhu, Chunxu Cao , Shuaishuai Zhang , Yan Zhang and Weisheng Zou .2011. pH-control modes in a 5-L stirred-tank bioreactor for cell biomass and exopolysaccharide production by Tremella fuciformis spore. Bioresource Technology. 102 :9175-9178.
34.Hirut O,Futamura,T.,Takebe, H., Satoh, A., Kamisaka, Y., Yokochi, T.,Nakahara, T.and Suzuki, O. 1996. Optimization and scale-up of γ-linolenic acid production by Mortie rella ramanniana MM15 a Highγ-linolenic acid producing mutant.J Frem Bioeng.82:366-370.
35.Hsu WK, Hsu TH, Lin FY, Chenga YK and Yanga JP. 2013.Separation, purification, and α-glucosidase inhibition of polysaccharides from Coriolus versicolor LH1 mycelia. Carbohydrate Polymers. 92:297-306.
36.Hecimovic I, Cvitanovic AB, Horzic D, Komes D.2011. Comparative study of polyphenols and caffeine in different coffee varieties affected by the degree of roasting. Food Chemistry.129: 991-1000.
37.Ko, H. J., Song, A., Lai, M.-N. and Ng, L.-T. 2009. Antioxidant and Antiradical Activities of Wu Ling Shen in a Cell Free System. The American Journal of Chinese Medicine. 37(04): 815-828.
38.Ko,H.J., Song,A., Lai,M.N., Ng, L.T. 2011.Immunomodulatory properties of Xylaria nigripes in peritoneal macrophage cells. Journal of Ethnopharmacology .138:762-768.
39.Liu, Q.N., Liu, R.S., Wang, Y.H., Mi, Z.Y., Li, D.S., Zhong, J.J.andTang, Y.J. 2009. Fed-batch fermentation of Tuber melanosporum for the hyperproduction of mycelia and bioactive Tuber polysaccharides. Bioresour Technol. 100:3644-3649.
40.Mao XB, Eksriwong T, Chauvatcharin S and Zhong JJ. 2005. Optimization of carbon source and carbon/nitrogen ratio for cordycepin production by submerged cultivation of medicinal mushroom Cordyceps militaris. Process Biochemistry. 40:1667-1672.
41.Ohno, N., Miura, N.N., Nakajima, M. and Yadomae, T. 2000. Antitumor 1,3-beta-glucan from cultured fruit body of Sparassis crispa. Biol. Pharm. Bull. 23: 866-872.
42.Ohno, N., Furukawa, M., Miura, N.N., Adachi, Y., Motoi, M. and Yadomae, T. 2001. Antitumor beta glucan from the cultured fruit body of Agaricus blazei. Biol. Pharm. Bull. 24: 820-828.
43.Sun, L J.,Wang, Y. P. and Li, Y. G. 2008. Study on fermented condidon of Cordyceps Sinensis. Science and technology of food industry.4:158-162.
44.Song, A., Ko, H. J., Lai, M. N. and Ng, L. T. 2011. Protective effects of Wu-Ling-Shen (Xylaria nigripes) on carbon tetrachloride-induced hepatotoxicity in mice. Immunopharmacology and Immunotoxicology. 33: 454-460.
45.Tang, Y.J., Zhang, W. and Zhong, J.J. 2009. Performance analyses of a pH-shift and DOTshift integrated fed-batch fermentation process for the production of ganoderic acid and Ganoderma polysaccharides by medicinal mushroom Ganoderma lucidum. Bioresour. Technol. 100:1852-1859.
46.Yang, F. C. and Liau. C.B .1998. Effect of cultivating conditions on the mycelia groth of Ganoderma lucidum in submerged flask cultures. Bioprocess Engineering. 19:233-236.
47.Zhong, J. J., Fang, Q. H. and Tang, Y. J. 2002. Enhanced production of valuable bioactive metabolites in submerged cultures of medicinal mushroom Ganoderma lucidum by manipulation of oxygen supply. J. Plant Biotechnol 4:109-115.



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