對氧磷是殺蟲劑巴拉松的主要代謝物，會干擾生物之肌肉反應，並在重要器官引發嚴重症狀，最終導致死亡。因此，本研究擬建構出利用啟動子調控來偵測對氧磷的全細胞感測器，然而，目前尚未發現藉由對氧磷進行誘導的啟動子，而研究室先前的研究中，有一質體含有酚類誘導轉錄因子dmpR，可偵測對氧磷降解後之對硝基苯酚（p-nitrophenol, p-NP）。此外，由於有機磷水解酶（organophosphorous hydrolase, OPH）的胞內表達仍存在一些問題，因此本研究以可大量分泌生物外膜囊泡（Outer membrane vesicles, OMVs）的E. coli JC8031為宿主，將OPH展示在OMVs表面，並結合酚類誘導型啟動子調控綠螢光蛋白，當環境中之對氧磷被OPH降解後，產物p-NP會誘導綠螢光基因的轉錄，以產生螢光訊號。我們已證明此模型可成功降解並感測環境中之對氧磷並發出螢光訊號。

另一感測系統中，由於環保意識抬頭加上天然資源有限的情況下，生質能成為了可替代能源的一個重要選項。生質能是由生物質所轉換而成的能量，而生物質則是來自植物和動物的可再生有機材料，其所儲存的化學能可通過各種過程轉化為我們可利用的能量，其中，反應條件溫和與操作簡單的生物轉化法逐漸受到重視。近年來，透過基因改殖技術將許多能分泌生物質水解酶之基因植入菌體中並大量表達，使其能於含有生物質的反應槽中直接進行水解，以節省酶純化等成本。然而，持續大量表達菌體本身非必要的基因可能導致菌體負荷量過大，進而導致菌體死亡。因此，本研究仿造了Clostridium thermocellum的染色體基因序列，設計一感測系統。當RsgI2跨膜蛋白感測到環境中之纖維素，即會釋放σI2轉錄因子，進而促使綠螢光蛋白之表達。此外，亦建構另一感測系統，當環境中存在纖維素時，即會被分泌至胞外的纖維素水解酶水解，產生水解產物纖維二糖後，誘導紅螢光蛋白之轉錄。目前已有初步的系統建立與測試結果，後續將持續調整系統結構以有效地進行纖維素的感測與水解。

As paraoxon is presented, it will lead to the cholinergic crisis in human beings. Therefore, we intend to construct a whole-cell biosensor to detect paraoxon. However, no promoter induced by paraoxon has been found yet. Luckily, there is a plasmid contained the phenol-inducible transcription factor, dmpR, in the previous research of our lab. Moreover, since there are still some problems in the intracellular expression of organophosphorous hydrolase (OPH). Thus, in this study, OPH was displayed on the outer membrane vehicles secreted by E. coli JC8031. Then, we combined with the plasmid contained a phenol-inducible promoter to regulate green fluorescent protein. When the paraoxon is degraded by OPH, the product p-nitrophenol will induce the transcription of green fluorescent gene. We have demonstrated that this model can successfully degrade and measure paraoxon in the environment. In another project, due to the limited natural resources, bioenergy has become a vital option for renewable energy. The chemical energy stored in biomass can be converted into energy that can be used by human through various processes. Among them, the biological method with mild conditions and simple operation has gradually attracted more and more attention. In recent years, bacteria capable of secreting biomass hydrolase have been added into reaction chamber. However, continuous expression of non-essential genes in bacteria may cause excessive load, leading bacteria to death. Therefore, we design two systems in order to sense and hydrolyze the cellulose. As far, the preliminary system establishment and test results have been obtained, and the system will be continuously adjusted to finally meet our purposes.

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