Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/52022
標題: 香杉芝菌絲體萃取物之組成、呈味性質及生物活性
Composition, taste quality and biological activities of extracts from mycelia of Antrodia salmonea
作者: 王麗諪
Wang, Li-Ting
關鍵字: 香杉芝;Antrodia salmonea;血管收縮轉換酵素;抗菌性質;抗發炎性質;angiotensin I-converting enzyme (ACE);antibacterial activity;anti-inflammation activity
出版社: 食品暨應用生物科技學系所
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摘要: 
香杉芝 (Antrodia salmonea T. T. Chang and W. N. Chou) 是寄生於香杉上的台灣特有真菌,常被誤認為樟芝 (Antrodia camphorata M. Zang and C. H. Su),學者張東柱與周文能於2004年鑑定為Antrodia屬之新種。近年來有學者分離出香杉芝之有效成分,發現其有抗發炎的功效以及對人類白血球細胞有抗氧化的能力,另外有研究從香杉芝菌絲體的氯仿萃取物中鑑定出具有抑制KB和HepG2腫瘤細胞株生長能力的有效成分。由於香杉芝文獻甚少,因此香杉芝的生理活性及有效成分值得進一步去探究,本實驗之研究目的為分析香杉芝菌絲體熱水以及乙醇萃取物的成分、呈味、生物活性物質,並探討對ACE抑制活性、抗菌性質以及抗發炎的能力。
經分析得知,香杉芝菌絲體熱水萃取物中,總糖及還原糖含量分別為405.60及108.58 mg/g,胜肽含量則為309.15 mg/g,乙醇萃取物中的總糖及還原糖含量則分別為183.92及158.06 mg/g,胜肽含量為391.96 mg/g;呈味物質分析方面,熱水及乙醇萃取物中的游離胺基酸總含量分別為220.69及254.59 mg/g;另外在生物活性物質方面,麥角硫因的含量分別為154.67及16.79 μg/g。
香杉芝菌絲體熱水及乙醇萃取物對ACE抑制活性方面,其IC50分別為1.42及1.27 mg/mL,乙醇萃取物的抑制ACE活性較佳;而ACEI抑制類型皆屬於混合型抑制;利用離心濃縮管,並不能有效提升熱水萃取物對ACE抑制之活性,而乙醇萃取物可由未經離心的93.71%提升到98.74-99.73%,但3000 ~ 50000 MWCO之抑制活性卻沒有差異。
在抗菌性質的部分,無論是香杉芝菌絲體熱水或乙醇萃取物,其抑菌圈直徑皆隨著樣品濃度增加而增加;萃取物經過濾之後,其抑菌能力降低;另外,在革蘭氏陽性菌當中,熱水萃取物對於B. cereus有最佳的抑菌效果,乙醇萃取物對於B. cereus以及L. monocytogenes有最佳的抑菌效果,對革蘭氏陰性菌來說,則皆以E. coli抑菌效果最佳;熱水萃取物對B. cereus的MIC及MBC皆為25 mg/mL,對E. coli的MIC及MBC皆為50 mg/mL,而乙醇萃取物對B. cereus以及L. monocytogenes之MIC和MBC則皆為6.25 mg/mL,對E. coli的MIC及MBC分別為6.25及12.50 mg/mL。
抗發炎性質評估方面,主要探討香杉芝菌絲體熱水及乙醇萃取物對於經LPS誘導巨噬細胞RAW 264.7發炎反應之影響。由結果可得知,萃取物在樣品濃度為15-20 μg/mL,即可有效抑制LPS誘導巨噬細胞分泌NO,熱水及乙醇萃取物的NO生成抑制率分別為43.32-53.68及54.40-77.83%;而在樣品濃度為5-20 μg/mL,可有效抑制LPS誘導巨噬細胞分泌TNF-α,且隨著樣品濃度的增加,TNF-α生成量隨之減少,熱水及乙醇萃取物的TNF-α生成抑制率分別為50.03-59.92及41.93-70.02%。
綜合上述結果,香杉芝菌絲體熱水及乙醇萃取物皆具有抑制ACE活性、抗菌以及抗發炎的能力,但其中的有效成分及作用機制尚不清楚,因此,未來可再進一步研究,分離純化出其有效成分,並探討其作用機制,以提升其價值和應用。

Antrodia salmonea (T. T. Chang and W. N. Chou) grows in the empty rotten trunk of Cunninghamia konishii in Taiwan, the fungus is a new species of the genus Antrodia as a basidiomycete first identified in 2004. This basidioma is similar to Antrodia camphorata (M. Zang and C. H. Su), which has been used as a folk medicine in Taiwan. Recent research has pointed out that, new compopents isolated from A. salmonea have antioxidative activity in human leukocytes and exhibit anti-inflammatory activities. In addition, the CHCl3 extract of the mycelium of A. salmonea exhibited cytotoxic activity against KB and HepG2 tumor cell lines. However, only limited information is reported in the literature about this fungus. The objectives of this study were to analyze the nutritional, taste and biological active components, and to investigate the effect of hot water (AsW) and ethanolic (AsE) extracts from mycelia of A. salmonea on angiotensin I-converting enzyme (ACE) inhibitory, antibacterial and anti-inflammation activities.
The results of AsW contained 405.60 and 108.58 mg/g of total sugars and reducing sugars, wheras AsE contained 183.92 and 158.06 mg/g of total sugars and reducing sugar, respectively.The content of peptides in AsW and AsE were 309.15 and 391.96mg/g, respectively. With regard to taste characteristics, the content of total free amino acids were 220.69 and 254.59 mg/g in AsW and AsE, respectively. With regard to biologically active components, AsW and AsE contained 154.67 and 16.79 μg/g of ergothioneine, respectively.
On the ACE inhibitory activity, IC50 value of ACE of AsW and AsE were 1.42 and 1.27 mg/mL, respectively. The kinetics on ACE-inhibition indicated that AsW and AsE showed the mixed-type inhibition. Using different membrane centrifuge filter devices, the ACE inhibitory activity of AsW could not be increased. The ACE inhibitory activity of AsE could be increased from 93.71% to 98.74-99.73%, but the ACE inhibitory activity remained the same.
On the antibacterial activity, as the concentration of AsW and AsE increased, the inhibition zone diameter increased. After filteration, antibacterial activity of AsW and AsE decreased. AsW and AsE had the better antibacterial activity against G (+) B. cereus and L. monocytogenes, respectively. For G (-), AsW and AsE both were effective against E. coli. Both MIC and MBC were 25 mg/mL for AsW against B. cereus, and both MIC and MBC of AsW against E. coli were 50 mg/mL. On AsE against B. cereus and L. monocytogenes, both MIC and MBC were 6.25 mg/mL, and those were 6.25 and 12.50 mg/mL against E. coli, respectively.
On the anti-inflammation activity, we investigated the effect of AsW and AsE on LPS-induced NO and TNF-α production in RAW 264.7 cell. When the concentrations were 15-20 μg/mL, AsW and AsE had an inhibition on LPS-induced NO production. The inhibition of NO production in AsW and AsW were 43.32-53.68 and 54.40-77.83%, respectively. The TNF-α production could be inhibitd when the AsW and AsE concentration were 5-20 μg/mL, and the amount of TNF-α produced increased with the concentration increased. The inhibition of TNF-α production in AsW and AsW were 50.03-59.92 and 41.93-70.02%, respectively.
Overall, AsW and AsE had ACE inhibitory, antibacterial and anti-inflammation activity. But the bioactive compound and mechanism were still unknown. Therefore, to isolate and purify the bioactive compounds and to study their mechanism of action are an another area of investigation.
URI: http://hdl.handle.net/11455/52022
其他識別: U0005-3006201115571800
Appears in Collections:食品暨應用生物科技學系

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