基于正交实验和响应面法优化石蜡降解混合菌的降解条件研究

Optimization of Degradation Conditions of Paraffin⁃Degrading Mixed Bacteria Based on Orthogonal Experiment and Response Surface Methodology

李阔 王慧 杜胜男 王卫强

Kuo LI Hui WANG Shengnan DU Weiqiang WANG

辽宁石油化工大学 石油天然气工程学院，辽宁 抚顺 113001

College of Petroleum Engineering，Liaoning Petrochemical University，Fushun Liaoning 113001，China

微生物作用于原油除蜡是一种高效简约的方法。为进一步提高微生物的石蜡降解率，混合两种菌株L和K配制了石蜡降解混合菌；通过正交实验和单因素实验确定了影响混合菌降解的因素（培养温度、培养基初始pH、盐质量分数、V（L菌）/V（K菌））；采用Box-Behnken方法设计四因素三水平实验及响应面优化，并建立数学模型，探究了混合菌降解石蜡的最佳条件。结果表明，4种单因素的影响大小顺序为V（L菌）/V（K菌）>培养温度>盐质量分数>培养基初始pH，其中一次项、二次项以及个别交互项对石蜡降解率影响显著；混合菌降解石蜡的最优条件：培养温度为37.2 ℃、培养基初始pH=7.3，培养基盐质量分数为1.2%，V（L菌）/V（K菌）=1.0∶1.6，石蜡降解率可达58.67%；混合菌在优化条件下作用于原油，生物降解率为41.38%，有良好的应用前景。

Microbial dewaxing of crude oil is an efficient and simple method. In order to further improve the paraffin degradation rate of microorganisms, two strains L and K were used to form paraffin degradation mixed bacteria, and their degradation conditions were optimized. The factors affecting the degradation of mixed bacteria (culture temperature, initial pH value of culture medium, salt concentration, and V（L strain ）/V（K strain）) were determined by orthogonal experiment and single factor experiment. Box-Behnken method was used to design four?factor three?level experiment and response surface optimization, and a mathematical model was established to explore the best conditions for the degradation of paraffin by mixed bacteria. The experimental results showed that the influence of four single factors is as follows: V（L strain）/V（K strain）>culture temperature>salt concentration>initial pH value of culture medium. Their primary term, secondary term and individual interaction term had significant influence on the paraffin degradation rate. The optimal experimental operating conditions for the mixed bacteria to degrade paraffin were determined as follows: culture temperature 37.2 ℃, initial pH value of culture medium 7.3, salt concentration of culture medium 1.2%, ratio of mixed bacteria V（L strain）/V（K strain）=1.0∶1.6, and paraffin degradation rate 58.67%. The mixed bacteria were used to act on crude oil under the optimized conditions, and the biodegradation rate was 41.38%,have good application prospects.

辽宁省高等学校创新人才支持计划项目(LR2018055);辽宁省自然科学基金指导计划项目(20180550745)

10.12422/j.issn.1672-6952.2024.05.004