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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	國立臺灣大學	申請系所	農業化學系
專案主持人	施養信	職等/職稱	教授
專案中文名稱	研發穩定型奈米釋氧顆粒以應用於受柴油汙染土壤之生物復育整治
中文關鍵字	穩定型奈米金屬釋氧顆粒、柴油、生物分解、界面活性劑、分子生物
專案英文名稱	Development of the stabilized oxygen-releasing nanoparticles to improve the bioremediation for the diesel oil-contaminated soil
英文關鍵字	stabilized oxygen-releasing nanoparticles, total petroleum hydrocarbon, bioremediation, PCR-DGGE, biosurfactant
執行金額	1,066,000元
執行期間	2015/11/17 至 2016/11/16
計畫中文摘要	本計畫的主要目的在於統整柴油污染場址生物整治之必要因素，創新開發具有整合多種生物刺激功能之「穩定型奈米金屬釋氧顆粒」添加劑，探討此添加劑對於柴油污染土壤及地下水生物降解之成效，嘗試提出未來可廣泛應用於柴油污染場址之釋氧顆粒，並搭配分子生物技術能有效監測土壤柴油降解微生物降解效能，以期研發低成本高效益的生物復育方式。 目前已備製多種過氧化鈣(Calcium peroxide, CaO2)，以動態光散射儀(dynamic light scattering, DLS)量測平均粒徑，其顆粒分布範圍主要在10-1,000 nm之間，PDI值為0.267，顯示透過研磨並添加分散劑後過氧化鈣顆粒可以有效的分散並不再聚集，其中以Wet-grinded-CaO2平均粒徑為最小(123.2 nm)。在柴油降解微生物方面，已篩選出一些具柴油降解能力之混菌，以Basic medium添加柴油(1,000 ppm) 35℃培養14天，與國內文獻報導之最佳柴油分解菌Gordonia alkanivorans (CC-JG39)相比較，去除效率分別為71.9% ( effective microorganisms, EM )及61.8%(CC-JG39)。土壤降解初步結果，以復育土所馴養微生物亦有柴油降解的情形，顯示原生菌群中已具有可分解柴油之微生物，經馴養後可經由降解實驗驗證其降解能力。在分子生物監測技術之建立上，利用聚合酵素鏈鎖反應(polymerase chain reaction, PCR)偵測使用的EM菌確實存在特定柴油降解基因，進一步配合變性梯度明膠電泳（denaturing gradient gel electrophoresis, DGGE）進行菌群結構分析，菌種豐富度隨復育天數上升之而後因主要碳源之柴油濃度下降後而菌種隨之下降。藉本土菌篩選出具有柴油降解之特殊菌群，並配合穩性性奈米過氧化鈣的處理，以及整合分子生物資訊的對應關係，希冀未來可提供學界、產業界與土壤及地下水整治的新策略。
計畫英文摘要	There were some problems in bioremediating the petroleum hydrocarbon contaminated soils, such as low contaminant degradation rate and limited degradation amounts. The objective of this project is to establish a cost-effectiveness technique for enhancing the efficiency of bioremediation by developing novel stabilized oxygen-releasing nanoparticles (SORNPs), which would mitigate the deficiency of microbial demands. On the other hand, genetic tool would also be developed to evaluate potentials of microorganisms in the biodegradation processs. We have synthesized several oxygen-releasing calcium peroxide (CaO2) particles by wet synthesis methods and grinding method (both wet or dry). Their particle sizes were ranged from 10-1,000 nm according to the measurement by dynamic light scattering (DLS) and transmission electron microscopy (TEM). The Wet-grinded CaO2 particle has the smallest in size (123.2 nm) and was most stable. The diesel degradation percentage of our mix culture (Effective Microorganisms, EM) was 71.9% compared to the 61.8% of the pure culture (CC-JG39) which the sampling point was at day 14 and the degradation experiments were performed at 35℃ using basic medium with 1,000 ppm of diesel as the main carbon source. The microorganisms in the mix culture also presented diesel degrading abilities in the treated soil. All these results indicated the possible presence of diesel degrading microorganisms in the mix culture. We further prove the existence of diesel degrading related genes and microorganisms in our mix culture using gene specific primers and denaturing gradient gel electrophoresis (DGGE).The DGGE data suggested the species in the consortium increased with the increase of carbon source during remediation and decreased with the depletion of carbon source. Our results demonstrated that the screened microbial consortium showing diesel degradation property could have great interest to academic and industry when combining its usage with our stable CaO2 nanoparticles in terms of diesel degradation in soil and under water remediation.