牛樟芝皿培技術之創新及其商品化應用

Innovation and Commercialization of Petri-Dish Producing Processes for Culturing Antrodia cinnamomea

戴瑞凡(Dai-Rui Fan)；傅詩涵(Shih-Han Fu)；顏宏愷(Hung-Kai Yen)

牛樟芝 ； 皿培技術 ； 三萜類 ； Antrodia cinnamomea ； Plate culture ； Dropping pills

美和學報

35卷1期（2016 / 05 / 01）

125 - 142

繁體中文

牛樟芝（Antrodia cinnamomea）為台灣特有真菌，具有多種活性成分，於研究中證實其萃取物具有抗發炎及抑制癌細胞生長的效果，然而其對於宿主專一性高，僅生長於台灣國寶牛樟樹(Cinnamomum kanehirae)，牛樟樹香味特殊且防腐效果好，於明清時其被大量砍伐製成雕刻品，導致現今野生牛樟芝已不多見。現今生活習性，人們不斷追尋珍稀的養生聖品，牛樟芝更是爭奪的對象，因此市場需求大。故開發出有效率的人工培養方式為重要目標，本研究中利用皿培技術將牛樟菌接種於培養基上，為期3個月的培養，並觀察其形態特徵以及利用分子 生物技術進行基源鑑定。每一批次之牛樟芝生長型態相似，且特定之基因序列相同，因此將皿培牛樟芝製成乾燥粉末，在進行乙醇萃取及減壓濃縮後，所獲得之萃取物以HPLC進行成分分析，並開發為滴九劑型進行商品化之雛形，將有助於提升牛樟芝成分穩定性及生物利用率。

Antrodia cinnamomea is an unique fungi in Taiwan. It has been proved that its extracts could against inflammation and inhibit the growing of cancer cells in many references. However, Cinnamomum kanehirae is a very special species in Taiwan. It has a special odor and can be used in antisepsis. Therefore, it had been felled a lot during Ming and Qing dynasties for sculpture, result in the disappearing of it. Nowadays, people increase their life style and pursue for rare health food, Cinnamomum kanehirae showed high marketing value in it. In consequently, to establish high effective artificial culture was an aim. In this study, the Antrodia cinnamomea fungus were inoculated by plate culture, and cultured for three months. The morphological characteristic and plant original identification were identified during culturing. While the growing habit in every batch is similar, and the DNA sequence is same, dried to mill powder, extracted by ethanol and vacuum concentration in order to get the extract and analyzed by HPLC, to develop dropping pills, contribute the stability and bioavailability of Antrodia cinnamomea.

1. Chang, C. W.,Chen, C. C.,Wu, M. J.,Chen, Y. S.,Chen, C. C.,Sheu, S. J.(2013).Active Component of Antrodia cinnamomea Mycelia Targeting Head and Neck Cancer Initiating Cells through Exaggerated Autophagic Cell Death.Evid Based Complement Alternat Med,2013
2. Chang, T. T.,Chou, W. N.(1995).Antrodia cinnamomea sp. nov. on Cinn amomum kanehirai in Taiwan.Mycological Research,99(6),756-758.
3. Chang, T. T.,Chou, W. N.(2004).Antrodia cinnamomea reconsidered and A. salmonea sp. nov. on Cunninghamia konishii in Taiwan.Botanical Bull etin of Academia Sinica,45(4),347-352.
4. Chen, C. C.,Shiao, Y. J.,Lin, R. D.,Shao, Y. Y.,Lai, M. N.,Lin, C. C.(2006).Neuroprotective diterpenes from the fruiting body of Antrodia camphorate.J. Nat. Prod.,69(4),689-91.
5. Chen, C. H.,Yang, S. W.,Shen, Y. C.(1995).New steroid acids from Antrodia cinnamomea, a fungal parasite of Cinnamomum micranthum.Journal of Natural Products,58(11),1655-1661.
6. Chen, Y. J.,Chou, C. J.,Chang, T. T.(2009).Compound MMH01 possesses toxicity against human leukemia and pancreatic cancer cells.Toxicol In Vitro,23(3),418-424.
7. Chen, Y. Y.,Chou, P. Y.,Chien, Y. C.,Wu, C. H.,Wu, T. S.,Sheu, M. J.(2012).Ethanol extracts of fruiting bodies of Antrodia cinnamomea exhi bit anti-migration action in human adenocarcinoma CL1-0 cells through the MAPK and PI3K/AKT signaling pathways.Phytomedicine,19(8-9),768-778.
8. Cherng, I. H.,Wu, D. P.,Chiang, H. C.(1996).Triterpenoids from Antrod ia cinnamome.Phytochemistry,41(1),263-267.
9. Dai, Y. Y.,Chuang, C. H.,Tsai, C. C.,Sio, H. M.,Huang, S. C.,Chen, J. C.,Hu, M. L.(2003).The Protection of Antrodia camphorata against Acute Hepatotoxicity.Journal of Food and Drug Analysis,11(3),177-185.
10. Geethangili, M.,Tzeng, Y. M.(2011).Review of pharmacological effects of Antrodia camphorata and its bioactive compounds.Evid Based Complement Alternat Med,2011
11. Hsu, F.-L.,Chou, C.-J.,Chang, Y.-C.,Chang, T.-T.,Lu, M.-K.(2006).Promotion of hyphal growth and underlying chemical changes in Antrodia camphorata by host factors from Cinnamomum camphora.International journal of food microbiology,106(1),32-38.
12. Hsu, Y. L.,Kuo, Y. C.,Kuo, P. L.,Ng, L. T.,Kuo, Y. H.,Lin, C. C.(2005).Apoptotic effects of extract from Antrodia camphorata fruiting bodies in human hepatocellular carcinoma cell lines.Cancer letters,221(1),77-89.
13. Hung-Chen, C.,De-Peng, W.,Cherng, I. W.,Chuen-Her, U.(1995).A ses quiterpene lactone, phenyl and biphenyl compounds from Antrodia cinnamo mea.Phytochemistry,39(3),613-616.
14. Ker, Y. B.,Peng, C. C.,Chang, W. L.,Chyau, C. C.,Peng, R. Y.(2014).Hepatoprotective bioactivity of the glycoprotein, antrodan, isolated from Antrodia cinnamomea mycelia.PloS one,9(4),1-11.
15. Lee, Y. P.,Tsai, W. C.,Ko, C. J.,Rao, Y. K.,Yang, C. R.,Chen, D. R.(2012).Anticancer effects of eleven triterpenoids derived from Antrodia camphorata.Anticancer Res.,32(7),2727-2734.
16. Lin, J.Y.,Wu, T.Z.,Chou, J.C.(2006).In vitro induction of fruiting body in Antrodia cinnamomea-a medicinally important fungus.Botanical Studies,47(3),267-272.
17. Liu, J. J.,Huang, T. S.,Hsu, M. L.,Chen, C. C.,Lin, W. S.,Lu, F. J.,Chang, W. H.(2004).Antitumor effects of the partially purified polysaccharides from Antrodia camphorata and the mechanism of its action.Toxicology and applied pharmacology,201(2),186-193.
18. Rao, Y. K.,Geethangili, M.,Tzeng, Y. M.(2013).Development of a high performance liquid chromatography method for the quantitative determination of bioactive triterpenoids in the extracts of Antrodia camphorata.Analytical Methods,5(20),5724-5730.
19. Shu, C. H.,Lung, M. Y.(2008).Effect of culture pH on the antioxidant properties of Antrodia camphorata in submerged culture.Journal of the Chinese Institute of Chemical Engineers,39(1),1-8.
20. Song, T. Y.,Yen, G. C.(2003).Protective effects of fermented filtrate from Antrodia camphorata in submerged culture against CCl4-induced hepatic toxicity in rats.Journal of Agricultural and Food Chemistry,51(6),1571-1577.
21. Song, T. Y.,Yen, G. C.(2002).Antioxidant properties of Antrodia camphorata in submerged culture.Journal of Agricultural and Food Chemistry,50(11),3322-3327.
22. Wu, S. H.,Ryvarden, L.(1997).Antrodia camphorata ("niu-chang-chih"), new combination of a medicinal fungus in Taiwan.Botanical Bulletin-Academia Sinica Taipei,38,273-276.
23. Wu, S. H.,Yu, Z. H.,Dai, Y. C.,Chen, C. T.,Su, C. H.,Chen, L. C.(2004).Taiwanofungus, a polypore new genus.Fungal Sci,19(3-4),109-116.
24. Wu, S. H.,Yu, Z. H.,Dai, Y. C.,Chen, C. T.,Su, C. H.,Chen, L. C.,Hsu, W. C.,Hwang, G. Y.(2004).Taiwanofungus, a polypore new genus.Fungal Sci,19(3-4),109-116.
25. Yang, S. W.,Shen, Y. C.,Chen, C. H.(1996).Steroids and triterpenoids of Antodia cinnamomea-A fungus parasitic on Cinnamomum micranthum.Phytochemistry,41(5),1389-1392.
26. Zang, M.,Su, Q. H.(1990).Ganoderma comphoratum, a new taxon in genus Ganoderma from Taiwan, China.Acta Botanica Yunnanica,12(4),395-396.
27. 朱祐頡(2008)。碩士論文(碩士論文)。台東，國立東華大學生物技術研究所。
28. 行政院衛生署中醫藥委員會(2000)。臺灣原住民藥用植物彙編：原住民藥用植物。台北:行政院衛生署中醫藥委員會。
29. 李建鈞(2009)。碩士論文(碩士論文)。台南，嘉南藥理科技大學藥物科技研究所。
30. 周廣麒、萬曉星、侯友松、張長鎧(2004)。蛹蟲草液態深層發酵的研究。食品與發酵工業，30(8)，39-43。
31. 凌士廷(2010)。碩士論文(碩士論文)。台中，朝陽科技大學生化科技研究所碩士班。
32. 張東柱(2009)。牛樟與牛樟芝的恩怨情仇。科學人，88，112-113。
33. 張家源(2014)。碩士論文(碩士論文)。台北，銘傳大學生物科技學系碩士班。
34. 劉崇良(2014)。碩士論文(碩士論文)。雲林，環球科技大學生物技術研究所。
35. 鄭維仁(2012)。碩士論文(碩士論文)。台南，南台科技大學生物科技系。
36. 簡秋源、姜宏哲、陳淑貞(1997)。牛樟菇培養性狀及其三帖類成分分析之研究。林業叢刊，72，133-137。