菌株P.aeruginosa BC1 的筛选、鉴定及其产生物表面活性剂的性能

Screening，Identification of Strain P. aeruginosa BC1 and Characterization of Its Biosurfactant

刘文磊 杨明明 张燕 陈春燕 肖泽仪

LIUWenlei YANG Mingming ZHANG Yan CHEN Chunyan XIAO Zeyi

为改善克拉玛依油田的微生物强化原油开采，从克拉玛依油田废水中筛选出一株性能优良的生物表面活性剂生产菌株BC1。根据菌株BC1 的理化指标和16S rDNA对其进行了属种信息鉴定，通过薄层层析（TLC）、傅里叶红外色谱（FT-IR）及高效液相色谱/质谱（HPLC-MS）对其产生物表面活性剂进行了分析，通过单因素摇瓶实验考察了发酵条件对菌株BC1 发酵合成生物表面活性剂的影响，研究了生物表面活性剂的性能。研究结果表明，菌株BC1 为铜绿假单胞菌属（Pseudomonas aeruginosa），其产生物表面活性剂主要成分为单鼠李糖脂RhC10C10；在温度37℃、初始pH为8、初始葡萄糖浓度5 g/L 的条件下持续发酵96 h 后，用酸沉降法测得鼠李糖脂产量为0.788 g/L；菌株BC1 产生物表面活性剂能将发酵液表面张力从72 降至28.6 mN/m，对车用机油的乳化指数E24 高达93%。人工油砂剥油实验中，发酵液、发酵离心上清液和鼠李糖脂粗品溶液对人工油砂的剥油率均达到74%以上。菌株P.aeruginosa BC1 产生物表面活性剂在微生物强化采油等方面具有良好的应用潜能。图7 表3参21

Strain BC1 with excellent performance of biosurfactant production was isolated from the wastewater of Karamay oilfield in order to improve the microbial enhanced oil recovery process. According to the physiological and biochemical indexes and 16S rDNA sequences analysis，the strain BC1 was classified. The biosurfactant produced by strain BC1 was identified by using thin-layer chromatography （TLC） ， Fourier transform infrared spectrum （FT-IR） and high performance liquid chromatography-mass spectrometry（HPLC-MS）analysis. The effect of fermentation conditions on the biosurfactant produce of strain BC1 was studied by single-factor shaking-flask culture experiment，and the performance of biosurfactant was researched. The results showed that strain BC1 belonged to Pseudomonas aeruginosa，and the key component of biosurfactant produced by strain BC1 was mono-rhamnolipid RhC10C10. The RLs yield was 0.788 g/L determined by acid precipitation method，under the condition of fermentation: 37℃ culture temperature，8 initial pH value，5 g/L glucose concentration，96 h fermentation time. The biosurfactant production of strain BC1 could dramatically reduce the broth surface tension from 72 to 28.6 mN/m，and show high emulsification index against motor oil of 93%. In the oil stripping test，more than 74% of crude oil could be effectively stripped from the artificial oil sands by using the fermentation broth，fermentation supernatant and solution of crude rhamnolipid. The biosurfactant produced by strain P.aeruginosa BC1 had good application potential in microbial enhanced oil recovery and other aspects.

10.19346/j.cnki.1000-4092.2016.04.030