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題 名 | Expression of a Novel Ethylene-producing Bifunctional Fusion Enzyme in Yeast=一種新的可產生乙烯之雙功能溶合酵素在酵母中之表現 |
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作 者 | 呂炳文; 於炳; 李凝; | 書刊名 | Botanical Bulletin of Academia Sinica |
卷 期 | 40:2 1999.04[民88.04] |
頁 次 | 頁107-114 |
分類號 | 368.87 |
關鍵詞 | ACC氧化酶; ACC合成酶; 雙功能; 乙烯; 溶合酶; 代謝工程; 報導基因; 酵母; ACC oxidase; ACC synthase; ACSO; Bifunctional; Ethylene; Fusion enzyme; Metabolic engineering; Reporter gene; Yeast; ACSO; |
語 文 | 英文(English) |
中文摘要 | 我們在酵母中重建高等植物中的通過1-羧酸 (ACC) 生物合成途徑,這個重建的通過ACC的乙生物物合成程是使一個新的有具功能的。由ACC合成庫〔〕和ACC氧化〔〕(ACS0)撥,合而成的溶合酸(Ning Li , Xian g Ning Jiamg Gno Ping cai and Shang Fa Yang[1996] The Journd of Biological Chemistry 27l:25738.2574l )所僵化。道個ACSO溶合酸彼再一次溶合到容〔〕甘?S-轉化。溶合後的ACSO溶合酸有把酸母內源腺?〔〕酸轉變成乙烯的能力。在酸母〔〕〔〕中的溶合酸GST-ACSO〔〕確定為110kDA。而含有GST-ACSO溶合酸的[]母細胞的乙生產速度,在半乳糖8小時的時侯,被確定為21.4 pmolowyi nc agdi owyi 10[]細胞。而ACC作為從腺[][]酸到乙的中間產物,在半乳糖誘導後呈逐增加,鑒于ACSO溶合酸能夠利用活的 真細胞中普遍存在的腺?〔〕〔〕酸生產乙,並且用來測量乙的去具有簡單、快速及極度〔〕〔〕 ( 百萬分之O.003) 的特點,我們預期這個[]功能的溶合酸在不久的將,來將會被應用於分子生物,發育生物,學發酸及達傳工程研之中。 |
英文摘要 | The 1-aminocyclopropane-1-carboxylic acid (ACC) dependent ethylene biosynthetic pathway of higher plants was reconstituted in yeast (Saccharomyces cerevisiae). The ACC-dependent ethylene biosynthesis in yeast was catalyzed by a novel bifunctional ACC synthase-ACC oxidase (ACSO) fusion enzyme (Ning Li, Xiang Ning Jiang, Guo Ping Cai and Shang Fa Yang [1996] The Journal of Biological Chemistry 271: 25738-25741). This fusion enzyme ACSO which was further fused to glutathione S-transferase is capable of converting yeast endogenous S-adenosyl-L-methionine (AdoMet) to ethylene. The molecular weight of the fusion enzyme, GST-ACSO, expressed in yeast, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), was 110 kDa. The ethylene production rate of the yeast cell containing GST-ACSO fusion enzyme was determined to be 21.4 pmol hr' per 108 cells at 8 h of galactose induction. The level of ACC, produced as an intermediate during the sequential reactions from AdoMet to ethylene, was found to increase gradually after galactose induction. Because ACSO is capable of producing ethylene from the ubiquitous and prevalent AdoMet in the living eukaryotic cell and the method commonly used to measure ethylene is simple, fast, and extremely sensitive (0.03 parts per billion), we anticipate this bifunctional fusion enzyme to be useful in the near future for research in molecular biology, developmental biology, fermentation, and genetic engineering. |
本系統中英文摘要資訊取自各篇刊載內容。