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基于正交试验和Box-Behnken试验的石油生物降解影响因素优化研究
文炜涛1, 王新伟1, 蔡婷1, 邹洪媛1, 杨永森2, 艾佳臻1, 肖萌3,4
0
(1.中国石油大学 北京 地球科学学院;2.中国石油大学(北京);3.中国地质大学北京水资源与环境学院,;4.中国地质大学北京水资源与环境学院)
摘要:
探索多种菌种降解石油过程中菌种和脂肽生物表面活性剂的作用,筛选石油降解主要因素及最佳组合,并为石油污染物的降解机理研究和石油污染修复提供指导。基于正交试验筛选主要影响因素,采用Box-Behnken试验探讨各因素最佳水平。正交实验中脂肽生物表面活性剂是多菌种降解石油的过程中最主要的影响因素,其在Box-Behnken试验中其能显著(P<0.05)的影响石油降解率。菌种降解能力是石油饱和烃组分生物降解的最主要影响因素,但脂肽生物表面活性剂是芳烃、胶质和沥青质组分降解的最主要的影响因素。研究所用菌种中,解淀粉芽孢杆菌(Bacillus amyloliquefaciens)和假单胞菌(Pseudomonas aeruginosa)在石油降解过程最重要,是本实验的石油降解最优菌。菌种和脂肽生物表面活性剂的添加浓度配比对于石油降解具有重要的影响。解淀粉芽孢杆菌和假单胞菌添加量5%(v/v),脂肽生物表面活性剂粗品添加量200 mg/L降解效果最好,理论情况下最高降解率可达63.78%,验证实验降解率达到了53.89%,相对于多菌种正交最好效果提高了5.54%。利用正交和Box-Behnken试验筛选最优降解菌和最优菌种复配具有分析因素多、实验量少等优点,具有较好的应用前景。
关键词:  脂肽生物表面活性剂  多菌种复配  正交实验  Box-Behnken实验
DOI:
投稿时间:2018-06-30修订日期:2018-11-14
基金项目:国家自然科学基金,生物表面活性剂对稠油重质组分生物降解屏障的作用机制(41403067)
Optimization of petroleum biodegradation factors based on orthogonal test and Box-Behnken test
Abstract:
There are many factors affect the biodegradation process of the oil, however, how the process affected by the factors is still difficulty and weak to research, this paper used the orthogonal test to select the main influencing factors, used Box-Behnken test to explore the optimal level of each factor, and established a multi-factor optimization method. Surfactin was the main factor for affecting the process of multi-bacteria degrading oil on orthogonal test, it could significantly affect the oil degradation rate (P<0.05) on Box-Behnken test. Species of the bacteria was the main factor affecting the biodegradation of saturate hydrocarbons. However, for the biodegradation of aromatic hydrocarbons, resins and asphaltenes, surfactin was the main factor. Among the ten of bacteria, Bacillus amyloliquefaciens and Pseudomonas aeruginosa played the most important role in oil degradation. In addition, the concentration ratio of bacteria and surfactin was also important for oil degradation. The most effective conditions were found to be 5% inoculation (v/v), and 200mg/L surfactin. Under the optimized condition, the maximum oil degradation rate can reach 63.78% in the theory conditions, the validation experimental degradation rate reached 53.89%, increased by 5.54% compared with the highest degradation rate on orthogonal experimental. There are some advantages to use orthogonal and Box-Behnken experiment to screen and combine the optimum oil-degrading bacteria, such as more factors to be analyzed and less experiment time, which will have a broad application prospect.
Key words:  surfactin  multiple strains  orthogonal test  Box-Behnken test.