查詢結果分析
相關文獻
- Fluorescence Versus Radioactivity for Assaying Antifungal Compound Inhibited Yeast 1,3-β-glucan Synthase Activity
- 宜蘭地區市售熟食肉品衛生指標菌之調查
- Screening and Cultivatin of a Yeast Strain with High Organoselenium Yields
- 以PCR之普遍性引子增殖混合酵母菌體的核醣體RNA基因之靈敏度
- 烘焙用酵母菌
- Cloning of a LEU Gene and an ARS Site of Rhodotorula glutinis
- 酵母菌的檢定及分類
- 苯甲酸用於蜜李最低用量之研究
- 李子酒製作用酵母菌之篩選及研究
- 酵母菌在食品及生物產業之應用
頁籤選單縮合
題 名 | Fluorescence Versus Radioactivity for Assaying Antifungal Compound Inhibited Yeast 1,3-β-glucan Synthase Activity=以螢光性與放射性方法測試抗黴藥物所抑制的酵母菌1,3-β-葡聚醣合成酶活性之比較 |
---|---|
作 者 | 柯源悌; 鄭暐宜; | 書刊名 | 藥物食品分析 |
卷 期 | 13:2 2005.06[民94.06] |
頁 次 | 頁184-191+196 |
分類號 | 341.95 |
關鍵詞 | 1,3-β-葡聚醣合成酶; 苯氨藍; 抗黴藥; 酵母菌; 1,3-β-glucan synthase; Aniline blue; Antifungal; Yeast; Echinocandin; |
語 文 | 英文(English) |
中文摘要 | 酵母菌1,3-ß-葡聚醣合成略(EC2.4.1.34)的活性利用螢光性染劑結合 的方法與傳統式的放射性方法來測量,並用於被echinocandin類的抗黴藥化合物作用的抑制性比較;在螢光法中,含有葡聚醣合成酶的胞漿膜微粒體首先與基質UDP-glucose反應形成葡聚醣產物,按著產物會被溶解,並與苯氨藍進行特異性結合,形成1,3-ß-葡聚醣-螢光子-複合物來報導酵素活性。酵素活性測試液成份對螢光的影響也進行測試,檢量線則以酵母菌葡聚醣為標準品來建立。葡聚醣合成酶的活性以這兩種方法進行比較,並使用lμg/μL相同濃度的胞漿膜微粒體反應,結果,酵素活性以放射法測量,會得到38.3μM UDP-glucose基質濃度參與合成反應,相較於螢光法,卻相當於得到262 μM葡聚時產物的生成,顯示出利用放射性方法會有低估生成物的現象,或 螢光法會有高估生成物的現象;雖然對此,抗黴藥pneumocandin A0的作用以螢光法呈現出其抑制的IC50值為1.25μM,卻和以放射性法所得抑制值lμM相近。本研究總結,放射性與螢光性兩種方法可以檢測出被echinocandin 類抗黴藥作用的相近抑制值,對於放射性方法中的可能限制性也有所討論。 |
英文摘要 | Yeast membrane (1,3)-β-glucan synthase (GS; EC 2.4.1.34) activity was assayed by a fluorescent dye-binding method and a conventional radioactivity method to compare its in vitro inhibition by the echinocandin-class antifungal compound. In the fluorescence method, GS-containing microsomal plasma membrane was first reacted with UDP-glucose substrate to form the 1,3-β-glucan product. The product was then solubilized and bound specifically with aniline blue to form a 1,3-β-glucan-luorochrome dye complex for reporting the GS activity. The effect of each component in the GS assay buffer on fluorescence was examined. A calibration curve was constructed using yeast glucan as standard. In addition, the GS activities using the same amount of 1 µg/µL microsomal concentration in the two assays were compared. The radioactivity assay showed 38.3 µM UDP-glucose incorporation and corresponded to the formation of equivalent 262 µM yeast glucan in the fluorescent assay. It indicated a possible underestimation of 1,3-β-glucan products in the radioactivity assay or an overestimation of the products in the fluorescence assay. However, pneumocandin A0 exhibited an IC50 value of 1.25 µM in the fluorescent assay closely comparable with that of 1 µM in the radioactivity assay. In summary, both the fluorescence and radioactivity tests generated similar data and pneumocandin inhibited GS. Possible limitations in the radioactivity assay were also discussed. |
本系統中英文摘要資訊取自各篇刊載內容。