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两株绿脓杆菌对石油污染土壤的修复作用
引用本文:黄磊,赵婷婷,贺赟,刘一琳,刘畅,靳大耀,贾晓强.两株绿脓杆菌对石油污染土壤的修复作用[J].生物工程学报,2017,33(6):957-967.
作者姓名:黄磊  赵婷婷  贺赟  刘一琳  刘畅  靳大耀  贾晓强
作者单位:天津大学 化工学院生物工程系 系统生物工程教育部重点实验室,天津 300072,天津大学 化工学院生物工程系 系统生物工程教育部重点实验室,天津 300072,天津大学 化工学院生物工程系 系统生物工程教育部重点实验室,天津 300072,天津大学 化工学院生物工程系 系统生物工程教育部重点实验室,天津 300072,天津大学 化工学院生物工程系 系统生物工程教育部重点实验室,天津 300072,天津大学 化工学院生物工程系 系统生物工程教育部重点实验室,天津 300072,天津大学 化工学院生物工程系 系统生物工程教育部重点实验室,天津 300072
基金项目:国家重点基础研究发展计划 (973计划) (No. 2014CB745100),国家自然科学基金 (No. 21576197),天津应用基础与前沿技术研究计划 (No. 14JCQNJC06700),天津蓬莱19-3漏油事故赔偿项目 (No. 19-3BC2014-03),天津重大科研计划 (No. 16YFXTSF00460) 资助。
摘    要:本文旨在研究环境条件下微生物对石油污染土壤的修复情况。从矿井周边土样定向筛选出两株绿脓杆菌,摇瓶降解实验发现,两菌混合培养10 d原油降解率达到95.67%,比单菌培养提高至少32%,即两菌对原油降解具有协同作用。根据降解实验结果制备了混合修复菌剂,并且人工构建石油污染场地,展开中试场地修复试验,模拟不同的操作条件下土壤中原油的降解情况。经60 d修复发现,添加了菌剂的场地,石油烃含量下降趋势明显,每克土壤中石油烃含量从初始的0.8%降至0.1%–0.3%,其中额外添加有机肥作为补充碳氮源的场地,总石油烃降解率最高,达到85.28%。而未添加菌剂的对照组石油烃含量仅减少25.85%。

关 键 词:石油污染,生物修复,混合菌剂,协同作用,生物表面活性剂,场地试验
收稿时间:2016/12/8 0:00:00

Bioremediation of oil-contaminated field by two Pseudomonas aeruginosa strains
Lei Huang,Tingting Zhao,Yun He,Yilin Liu,Chang Liu,Dayao Jin and Xiaoqiang Jia.Bioremediation of oil-contaminated field by two Pseudomonas aeruginosa strains[J].Chinese Journal of Biotechnology,2017,33(6):957-967.
Authors:Lei Huang  Tingting Zhao  Yun He  Yilin Liu  Chang Liu  Dayao Jin and Xiaoqiang Jia
Institution:Key Laboratory of Systems Bioengineering (Ministry of Education), Department of Biological Engineering School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China,Key Laboratory of Systems Bioengineering (Ministry of Education), Department of Biological Engineering School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China,Key Laboratory of Systems Bioengineering (Ministry of Education), Department of Biological Engineering School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China,Key Laboratory of Systems Bioengineering (Ministry of Education), Department of Biological Engineering School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China,Key Laboratory of Systems Bioengineering (Ministry of Education), Department of Biological Engineering School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China,Key Laboratory of Systems Bioengineering (Ministry of Education), Department of Biological Engineering School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China and Key Laboratory of Systems Bioengineering (Ministry of Education), Department of Biological Engineering School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China
Abstract:STK1 is an important MAPK gene regulating the conidial development, osmotic stress and pathogenicity of Setosphaeria turcica. First, we constructed Pichia pastoris GS115 expression vector pPIC3.5K-EGFP containing enhanced green fluorescent protein gene (EGFP). Then, we amplified STK1 gene by PCR with the template of cDNA of S. turcica model isolate 01-23, and cloned into the vector pPIC3.5K-EGFP with enhanced green fluorescent protein gene (EGFP) to construct the STK1-EGFP fusion gene expression vector pPIC3.5K-STK1-EGFP. The vector was transformed into the susceptible cells of P. pastoris GS115 by electric shock process, and the transformants were identified by MD medium screening, PCR determination. STK1 gene and EGFP gene could be expressed effectively and stably in the transformants as detected by RT-PCR and fluorescence observation. In addition, we also found that the Kozak sequence before the start codon of STK1 gene could increase 4.8 folds expression level of STK1- EGFP fusion gene. The above research results laid a good foundation for subcellular localization and antibody preparation of STK1 protein.
Keywords:oil pollution  bioremediation  mixed bacterium agent  synergistic effect  biosurfactant  field test
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