采用酶液活性测定法快速筛选及构建石油烃高效降解菌系

快速筛选复杂有机物降解微生物混合菌系,在污染物治理过程中具有重要的实践意义.本研究首次尝试利用MicroResp^TM技术分析微生物酶液活性的方法,快速标定高效降解菌及混合菌系的石油烃降解能力,并采用传统的摇瓶培养检测法予以验证.通过微生物胞内、胞外及混合酶液的活性分析,考察了不同酶系(胞外、胞内及混合酶液)、菌系对石油烃分子的降解情况.结果表明: 结合MicroResp^TM技术的酶液活性测定法能够快速检测石油烃代谢酶系的降解能力,其灵敏度好、通量高,与传统的菌株摇瓶培养方法的检测结果基本一致.其中,7株菌株的120种全组合菌系活性测定试验在12 h周期内1次完成.筛选周期由传统摇瓶培养所需的7 d缩短10倍以上.以酶活性测定结果为指导设计的复配菌系具有较高的降解效率,最高石油烃降解率达(56.1±1.6)%.表明本筛选方法精度高、通量高,可用于石油烃降解功能菌系的构建.

Rapid screening and construction of high-efficient petroleum-degrading microflora through enzyme activity assay

It is of great practical significance to rapidly screen the mixed microbial flora for degradation of complex organic pollutants. It was the first time to rapidly calibrate the petroleum hydrocarbon degradation ability of highly efficient degrading microflora by measuring enzyme activity analyzed by the MicroResp^TM technology. The results were validated by the traditional shaking culture. According to the activity analysis of exoenzymes, endoenzymes and mixed enzymes, the degradation abilities of microflora and enzymes (including exoenzymes, endoenzymes and mixed enzymes) on petroleum hydrocarbon was investigated. The results showed that the enzyme activity assay had perfect sensitivity and high-throughput for rapidly detecting the degradation ability of the petroleum hydrocarbon. The test period was 12 h which was 10 times shorter than shaking culture for 120 microflora combinations. The maximum degradation extent of the petroleum hydrocarbon by microflora reached (56.1±1.6)%. It was concluded that the method of enzyme activity assay was high-efficient and high-throughput screening, which could be used in the construction of petroleum-degrading microflora.