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年度	105	專案性質	實驗性質
專案類別	研究專案	研究主題	整治
申請機構	國立中央大學	申請系所	生命科學系
專案主持人	陳師慶	職等/職稱	教授
專案中文名稱	開發高效能螯合與還原毒性汞之菌株及量產移汞蛋白質-metallothionein
中文關鍵字	汞,生物復育,微生物相,基因工程菌,metallothion
專案英文名稱	Application of the bacteria with highly potential of reducing and chelating mercury and/or chelating-mercury metallthionein on mercury-contaminated soil
英文關鍵字	mercury,bioremediation,genetic bacteria,metallothionein
執行金額	849,750元
執行期間	2015/11/17 至 2016/11/16
計畫中文摘要	土壤和水是地球上最重要的自然資源，然而，在過去數十年中，由於工業化、 人口的大量增加和城市化，環境重金屬汙染問題日益嚴重。其中，汞因其具高毒 性及生物累積等特性，可能對生物體及環境造成影響或危害，且具有高風險，因 此汞污染需優先被進行整治。利用生物復育程序(Bioremediation)進行環境污染整 治是屬於對環境最友善的技術，而且也具有包括經濟效益所需花費少，容易操作 且較易受社會大眾接受等優點。但由於汞與土壤形成之錯合物溶解度非常低，不 容易被生物所利用，因此藉由使用螯合劑輔助使土壤中汞錯合物由固相轉移至液 相，使生物能更有效處理汙染物為有效可行之方法，本研究團隊於去年研究計畫 已有初步成果。在期中報告的研究執行內容與成果我們著重在利用基因工程方式 建構強化先前研究內容菌種，開發具有潛力之汞抗性基因重組菌株，欲強化其產 生之汞抗性蛋白並探討未來應用模場整治可行性。 本次的期末報告，我們設計了新的穿梭載體pJBR 系列，使用持續性啟動子 J23100(不需使用誘導物)、核醣體結合位B0034以及rrnB T1終止位構成，並且能 讓MT-1和EGFP 的融合蛋白質順利的表現，之後，我們利用生物反應器(Bioreactor) 測試了轉型pJBME 質體的Pseudomonas A46菌株在靜止細胞培養液和生長細胞 培養液中固定汞的效果，在靜止細胞培養液中此株菌並沒有特別的提升固定汞能 力，但是在生長細胞培養液中，細胞中固汞量有明顯增加。本計畫成功開發抗汞 並固定汞之基因工程菌，與去年計畫淋洗法共同應用，未來則能在環境復育上開 闢新的前景。
計畫英文摘要	Soil and water are the most important natural resources on earth. However, environmental pollution of heavy metal is escalating due to industrialization, huge population and urbanization. Mercury has been reported as highly toxic and need priority treat. Bioremediation is economy and environmental friendly approaches and also acceptable by general public. However, due to very low solubility of mercury compounds. It’s not easy to be used by microbes. Therefore, using chelating agent to make mercury compound in soil transferred from solid phase to liquid phase is an effective way for bioremediation. We have isolated various strains of microorganisms from the previous study funded by “Soil and Groundwater Remediation Fund Management Board, Environmental Protection Administration, Executive Yuan” – “Screening and study of indigenous microorganisms from mercury contaminated soil for bioremediation application”. These indigenous microorganisms will be excellent candidates to develop bioremediation approach for mercury contaminated area in situ. In the final report, we design a novel shuttle vector – pJBR family, which consist of promoter J23100 and RBS B0034 and rrnB T1 terminator. This vector is very successful to let MT-1-EGFP fusion protein express in Pseudomonas A46. We also use the Bioreactor to test the mercury resistant ability of the Pseudomonas A46 which transfect with pJBME in the resting medium and growth medium. However, the resting medium cells didn’t show the enhancement ability of mercury resistance, but the growth cells indeed arise the mercury accumulation in the cell pellet. So that our plant is confirm to success, and we can also combine with the previous ground wash study method to fulfill the bioremediation application, it has a very good prospect in the future.