Zhang Heng, Yang Chao, Zhao Qiao, Qiao Chuanling
State Key Laboratory of Integrated Management of Pest Insects and Rodents, Institute of Zoology, Chinese Academy of Sciences, Beijing, China.
Bioresour Technol. 2009 Jul;100(13):3199-204. doi: 10.1016/j.biortech.2009.02.008. Epub 2009 Mar 9.
Simultaneous biodegradation of hexachlorocyclohexane (HCH) and organophosphates (OPs) by a recombinant Stenotrophomonas sp. was studied in the study. The broad-host-range plasmid pVGAB, harboring enhanced green fluorescent protein gene (egfp) and dehalogenase genes (linA and linB), was constructed and transformed into the OP-degrading strain Stenotrophomonas YC-1 to get the recombinant strain YC-H. Over-expression of dehalogenase (LinA and LinB) and enhanced green fluorescent protein (EGFP) was obtained in YC-H by determining their enzymatic activities and fluorescence intensity. YC-H was capable of rapidly and simultaneously degrading 10mg/l gamma-HCH and 100mg/l methyl parathion (MP) determined by GC-ECD analysis. A bioremediation assay with YC-H inoculated into fumigated and nonfumigated soil showed that both 10mg/kg gamma-HCH and 100mg/kg MP could be completely degraded within 32 days. The novel EGFP-marked bacterium could be potentially applied in the field-scale decontamination of HCH and OPs residues in the environment.
本研究对重组嗜麦芽窄食单胞菌同时生物降解六氯环己烷（HCH）和有机磷酸酯（OPs）进行了研究。构建了携带增强型绿色荧光蛋白基因（egfp）和脱卤酶基因（linA和linB）的广宿主范围质粒pVGAB，并将其转化到降解OPs的嗜麦芽窄食单胞菌菌株YC-1中，得到重组菌株YC-H。通过测定其酶活性和荧光强度，在YC-H中实现了脱卤酶（LinA和LinB）和增强型绿色荧光蛋白（EGFP）的过表达。通过气相色谱 - 电子捕获检测器（GC-ECD）分析测定，YC-H能够快速同时降解10mg/l的γ-HCH和100mg/l的甲基对硫磷（MP）。将YC-H接种到熏蒸和未熏蒸土壤中的生物修复试验表明，10mg/kg的γ-HCH和100mg/kg的MP均可在32天内完全降解。这种新型的EGFP标记细菌可能在环境中HCH和OPs残留的现场规模去污中得到应用。
