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具有高效降解地塞米松能力的伯克霍尔德菌CQ001株的分离及功能基因组学分析
Isolation and functional genomic analysis of a strain of
Burkholderia CQ001 degrading dexamethasone- 第三军医大学学报 2017年39卷第13期 页码：1352-1359
- 作者机构：
1.400016 重庆, 重庆医科大学基础医学院病原生物学教研室
2.400016 重庆, 重庆医科大学药学实验教学中心
3.400016 重庆，重庆医科大学基础医学院免疫学教研室
- 作者简介：
[ 王毅，E-mail: 421355043@qq.com WANG Yi, E-mail:421355043@qq.com
- 基金信息：重庆市渝中区科委科技项目(20160110)
DOI：10.16016/j.1000-5404.201612198
中图分类号： R123.3;R378.99;R394-33收稿：2016-12-28，
修回：2017-03-03，
纸质出版：2017-07-15
- 稿件说明：
移动端阅览
斯丹, 杨致邦, 蒋任举, 等. 具有高效降解地塞米松能力的伯克霍尔德菌CQ001株的分离及功能基因组学分析[J]. 第三军医大学学报, 2017,39(13):1352-1359.
Dan SI, Zhibang YANG, Renju JANG, et al. Isolation and functional genomic analysis of a strain of
Burkholderia CQ001 degrading dexamethasone[J]. Journal of Third Military Medical University, 2017, 39(13): 1352-1359.斯丹, 杨致邦, 蒋任举, 等. 具有高效降解地塞米松能力的伯克霍尔德菌CQ001株的分离及功能基因组学分析[J]. 第三军医大学学报, 2017,39(13):1352-1359. DOI： 10.16016/j.1000-5404.201612198.
Dan SI, Zhibang YANG, Renju JANG, et al. Isolation and functional genomic analysis of a strain of
Burkholderia CQ001 degrading dexamethasone[J]. Journal of Third Military Medical University, 2017, 39(13): 1352-1359. DOI： 10.16016/j.1000-5404.201612198.
摘要
目的
2
研究细菌对地塞米松类甾体激素的降解作用及其分子机制，为构建清除水环境医源性地塞米松类药物污染的工程菌奠定基础。
方法
2
采用富集培养法从医院废水中分离降解地塞米松的细菌，采用固相萃取-高效液相色谱(HPLC)法检测细菌对地塞米松的降解效能
通过16S rDNA序列测定进行鉴定。用Illumina Hiseq4000结合第三代测序技术对降解菌的全基因组测序，并进行序列组装、注释和分析，对降解地塞米松相关基因进行RT-qPCR验证。
结果
2
分离到1株对地塞米松有较高降解作用的细菌，经鉴定属于伯克霍尔德菌属(
Burkholderia
)，命名为
Burkholderia
sp.CQ001。该菌对地塞米松磷酸钠及地塞米松的降解率分别为84.8%和77.11%。全基因组测序表明
Burkholderia
sp.CQ001包含2条染色体和4个巨型质粒，与代谢相关基因大部分集中在2号染色体上，共3 260 157 bp。生物信息学分析表明，该菌含有甾体代谢通路中许多重要酶类的编码基因，其中与地塞米松降解相关的有ABC转运子，3-甾酮-9α-脱氢酶等，这些基因在以地塞米松磷酸钠为碳源的培养条件下表达量不同程度地高于以蔗糖为碳源的培养条件。
结论
2
Burkholderia
sp.CQ001是一株具有强大的代谢功能和代谢途径丰富的细菌，具有降解地塞米松类甾体激素的性能，该菌株为后续研究甾体激素的降解机制和构建清除水环境医源性地塞米松类药物污染的工程菌提供了菌种。
Abstract
Objective
2
To investigate the degradation effect and molecular mechanism of bacteria on dexamethasone steroid hormones
in order to lay the foundation for the construction of engineering bacteria to eliminate the pollution of iatrogenic dexamethasone drugs in water environment.
Methods
2
By enrichment culture method
dexamethasone degradation bacteria were isolated from hospital waste water. The degradation rates of dexamethasone were determined by using high performance liquid chromatography (HPLC). The dexamethasone degradation bacterium was identified with 16S rDNA sequencing. The whole genome sequencing was performed using Illumina Hiseq4000 combined with third generation sequencing technology
and sequence assembly
annotation and analysis. Some degradation of dexamethasone related genes were further verified by real time RT-qPCR.
Results
2
A strain of bacteria with higher degradation to d
examethasone were isolated
and then identified as
Burkholderia
which was named
Burkholderia
sp.CQ001. The degradation rates were 84.8% to dexamethasone sodium phosphate and 77.11% to dexamethasone. Genome sequencing results demonstrated that the bacterium contained 2 chromosomes and 4 giant plasmids. Most of the metabolic related genes were concentrated in chromosome 2
accounting for 3 260 157 bp. Bioinformatic analysis results showed that
Burkholderia
sp.CQ001 had some significant genes that encoded enzymes working in the steroid degradation pathways
including asABC transporter
3-ketosteroid-9alpha-monooxygenase
etc
which are involved in the dexamethasone degradation. The expression levels of these genes were higher when
Burkholderia
sp.CQ001was grown in A medium with dexamethasone sodium phosphate as the unique carbon source than in A medium with glucose as the unique carbon source.
Conclusion
2
Burkholderia
sp.CQ001 is A strain of bacterium with strong metabolic function and metabolic pathway. It has the properties of degrading dexamethasone steroid hormones. Our findings may provide new strain and much evidence for the further studies concerning dexamethasone degradation mechanisms and establishment of bioremediation engineering bacteria for eliminating dexamethasone pollution.
关键词
Keywords
references
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