Surfactin與rhamnolipid乃兩種不同結構的生物界面活性劑。由本研究中可得到，其二者臨界微胞濃度critical micelle concentration (CMC)相當低，surfactin為35 mg/L；rhamnolipid為52 mg/L。且此兩種生物界面活性劑對柴油乳化能力很強，surfactin在濃度為120 mg/L時可造成柴油58 %的乳化；而rhamnolipid則可在濃度為48 mg/L時即可使柴油62 %乳化。由於以上兩項重要的特性，因此本研究將以此兩種生物界面活性劑添加，探討生物界面活性劑添加濃度、環境中銨氮濃度及pH值，對柴油生物降解效率的影響。

　　在surfactin添加濃度方面，在40 mg/L時微生物有最佳的生長情況，比生長速率由未添加的0.041/hr提升至0.055/hr；在40 mg/L組別柴油降解速率可由未添加時20.1 mg TPH-d/L/hr大幅提升至86.7 mg TPH-d/L/hr，提升效果高達四倍。但高濃度的surfactin可在此實驗中明顯看出對微生物生長的抑制，在400 mg/L組別之比生長速率大幅下降至0.009/hr。不同的rhamnolipid添加濃度之下，微生物的生長曲線以添加濃度80及160 mg/L表現最佳，微生物之比生長速率由未添加0.021/hr 提升至0.047/hr；柴油降解速率較未添加20.5 mg TPH-d/L/hr 提升至137.3 mg TPH-d/L/hr，提升效果約為七倍。

　　pH對柴油生物降解影響方面，未添加生物界面活性劑時，柴油的降解速率以pH 7.2最佳(63.7 mg TPH-d/L/hr)，pH過高或過低皆會導致柴油降解速率下降至40.1 mg TPH-d/L/hr (pH 8.4)及19.7 mg TPH-d/L/hr (pH 5.2)。而當實驗中添加surfactin後微生物受抑制者為pH 5.2，微生物比生長速率從最高0.051/hr (pH 8.4)大幅下降至0.013/hr。pH 5.2時，柴油降解速率也從最高91.7 mg TPH-d /L/hr (pH 8.4)大幅下降至7.2 mg TPH-d/L/hr。而實驗中添加rhamnolipid後柴油降解速率方面最佳表現為pH 6.3-8.4，被抑制者為pH 5.2，柴油降解速率由最高85.7 mg TPH-d/L/hr (pH 7.4)下降至36.2。

　　環境中銨氮濃度對柴油生物降解影響方面，當未添加生物界面活性劑時，無論是微生物比生長速率還是柴油降解速率皆在銨氮濃度200-300 mg/L會有最佳的表現。而當實驗添加surfactin(40 mg/L)後，在銨氮濃度超過200 mg/L後微生物比生長速率高達0.057/hr；而當溶液中銨氮濃度不足時 (50 mg/L)微生物比生長速率只有0.035/hr。在添加rhamnolipid組別(50 mg/L)，柴油降解速率在銨氮濃度100-300 mg/L表現皆大同小異，約為52.1-65.4 mg TPH-d/L/hr；但當銨氮濃度升至450 mg/L時柴油降解速率便滑落至9.2 mg TPH-d/L/hr。

　　在柴油汙染土壤添加生物界面活性劑生物復育試驗中，發現無論是添加surfactin或添加rhamnolipid組別土壤在實驗進行達68天後土壤中柴油殘餘量皆可明顯比控制組來的低。surfactin(40 mg/kg)組土壤中柴油殘餘量約剩餘1655 mg TPH-d/kg；而rhamnolipid(50 mg/kg)組別土壤中殘餘柴油量更是低於250 mg TPH-d/kg。在此時控制組中土壤柴油殘餘量約為4678 mg TPH-d/kg。土壤中初始柴油濃度為6981 mg TPH-d/kg。

　　在分子生物檢測(DGGE)方面發現添加rhamnolipid組，菌相明顯較控制組和surfactin組為豐富。且由Marker比對中可得到，無論是控制組或實驗組(添加surfactin、rhamnolipid)的土壤中皆含有可分解柴油的菌株。

　　Surfactin and rhamnolipid are two types of biosurfactants with different chemical structures. In this study, we found that both of them owned relatively low critical micelle concentrations (CMC), which were 35 and 52 mg/L for surfactin and rhamnolipid, respectively. Also, both of the two biosurfactants can achieve significantly high emulsification index (E24) which resulted in the emulsification of diesel oils. The addition of 120 mg/L surfactin achieved E24 of 58%, which resulted in the emulsification of 58% diesel oil. However, only 48 mg/L rhamnolipid was needed to achieve the E24 (62%), and 62% of diesel oil was emulsified. Based on the two important characters, relatively CMC and high E24, this study focused on 1) the addition concentrations of the two biosurfactants, 2) ammonia concentrations, and 3) pH values in the environment, in order to investigate the effect of biosurfactants addition on diesel oil biodegradation.

　　In the study of adding different concentrations of biosurfactant, when adding different concentrations of surfactin to the diesel contaminated soil solution, the optimal microbial growth rate was attained with 40 mg/L of surfactin. The specific growth rate was increased from 0.041/hr to 0.055/hr, compared with the non-addition group. When adding 40 mg/L of surfactin, the degradation rate was raised from 20.1 mg/L/hr, with the non-addition group, to 86.7 mg/L/hr, which were about four-time improvement in the degradation rates. In spite of this, addition of high concentration of surfactin was found to cause an inhibition of the microbial activity. The specific growth rate dropped to 0.009/hr when 400 mg/L of surfactin was added. When adding different concentrations of rhamnolipid, the microbial growth curve indicated that an optimal bioactivity can be achieved with 80 or 160 mg/L of rhamnolipid addition. Compared with the non-addition group, the specific growth rate was increased from 0.021/hr to 0.047/hr, and the diesel degradation rate was raised from 20.5 to 137.3 mg TPH-d/L/hr. An approximately seven-fold enhancement of the diesel degradation was achieved.

　　In the study of pH investigation, with no addition of the biosurfactant, the optimal diesel degradation was 63.7 mg TPH-d/L/hr when pH value was 7.2. Extreme high (8.4) or low (5.2) pH values resulted in relatively low diesel degradation rate, 40.1 and 19.7 mg TPH-d/L/hr, respectively. With the addition of surfactin, at the pH value of 8.4, both of the specific growth rate and the diesel degradation rate reached the optimal. At the pH value of 5.2, the specific growth rate dropped from 0.051/hr (pH 8.4) to 0.013/hr; and the diesel degradation also decreased from 91.7 (pH 8.4) to 7.2 TPH-d mg/L/hr. Similarly, with the addition of rhamnolipid, the highest diesel degradation rate was attained when the pH values were around 6.3-8.4. At pH 5.2, the diesel degradation rate dropped from 85.7 (pH 7.4) to 36.2 TPH-d mg/L/hr。

　　In the study of ammonia investigation, with no addition of the biosurfactant, the optimal growth rate and diesel degradation rate were found with the ammonia concentrations of 200-300 mg/L. When adding 40 mg/L of surfactin, the specific microbial growth rate could achieve 0.057/hr with the ammonia concentration f 200 mg/L. Yet, the specific growth rate decreased to 0.035/hr when ammonia concentration was only 50 mg/L. When adding 50 mg/L of rhamnolipid, about equivalent diesel degradation (52.1-65.4 TPH-d mg/L/hr) was reached with the ammonia concentration of 100-300 mg/L. However, when 450 mg/L of ammonia concentration was provided, diesel degradation became significantly low (9.2 mg TPH-d/L/hr).

　　In the study of adding biosurfactants to diesel-contaminated soil, it was observed that, with the addition of either surfactin or rhamnolipid, the remained diesel was apparently lower than that in the control set after 68 days operation. With the addition of surfactin (40 mg/kg), the remained diesel in the soil was 1655 TPH-d mg/kg. With the addition of rhamnolipid (50 mg/L), the remained diesel was only lower than 250 mg TPH-d/kg. At the same time, the remained diesel in the control set was around 4678 TPH-d mg/kg which was originally 6981 TPH-d mg/kg.

　　In the study of molecular biomonitoring with DGGE, it concluded that with the addition of rhamnolipid, the microbial community was more diverse than that in the control set and in the surfactin addition. Also, in the result of Marker comparison, it indicated the existence of diesel-degrading bacteria in both the experimental and the control sets.

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