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中國畜牧學會會誌

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篇名 伏馬鐮胞毒素降解酵素之基因選殖
卷期 46增刊
並列篇名 Cloning of fumonisin-degrading enzyme
作者 黃雅婕游玉祥鄭永祥
頁次 223-223
關鍵字 伏馬鐮胞毒素基因選殖酵母菌Fumonisin BGene cloningYeast
出刊日期 201712

中文摘要

黴菌毒素是真菌產生的次級代謝物,伏馬鐮胞毒素(fumonisin B1, FB1)為經濟動物飼料中 常見的黴菌毒素,FB1 與神經鞘脂蛋白(sphingolipid)的結構相似,因此具干擾神經醯胺 合成酶(ceramide synthase)之活性,進而使細胞內累積神經鞘氨醇及鞘氨醇的含量。豬 隻採食受到 FB1 汙染的飼料會造成肺水腫並引發二次感染。本試驗目的為選殖伏馬鐮胞 毒素降解酵素之基因並用於酵母菌重組蛋白表現系統,伏馬鐮胞毒素降解酵素基因片段源 至鞘脂單胞菌中,將構築於酵母菌表現載體中,此表現載體具有易於培養和可高量表現重 組蛋白的特性。目前已成功地將此基因構築於兩種酵母菌表現載體中,並經定序確認基因 正確性且嵌入酵母菌載體。進一步以電破法(electroporation)將此載體送入酵母菌基因組 內,經電泳分析和抗生素篩選後,結果顯示電破後獲得正確菌種的成功率可達七成以上, 因此未來可用此酵母菌表現系統分泌伏馬鐮胞毒素降解酵素之重組蛋白,進一步開發作為 降解黴菌毒素之飼料添加劑。

英文摘要

Mycotoxins are secondary fungal metabolites and fumonisin B1 (FB1) is a common mycotoxin in animal feed. The structure of FB1 is similar to sphingolipids. Thus, FB1 is able to affect endogenous ceramide synthase activity, leading to intracellular accumulation of toxic sphingoid bases. FB1-contaminated feed could cause pulmonary edema and secondary infections in pigs. The purpose of the study is designed to clone a fumonisin-degrading enzyme gene. Here, we successfully cloned the fumonisin-degrading enzyme gene derived from sphingopyxis and ligated into two yeast expressing vectors. We also confirmed the DNA sequence of yeast expressing vectors by sequencing. The expressing vectors were then electroporated into the yeast genome and examined the recombinant efficiency by anti-antibiotic selection and agarose electrophoresis. Present results showed the recombinant efficiency of expressing vector in yeast genome was up to 70%. In the future, it is suitable for use on an industrial scale to produce large amounts of fumonisin-degrading enzyme by yeast expressing system. The product could be used as animal feed additive for prevention of mycotoxin contamination in feed.

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