Construction of malate-sensing Escherichia coli by introduction of a novel chimeric two-component system
Construction of malate-sensing Escherichia coli by introduction of a novel chimeric two-component system
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DOI：
10.1007/s00449-014-1321-3
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发表时间：
2015-04-01
影响因子：
3.8
通讯作者：
Hong, Soon Ho
中科院分区：
文献类型：
--
作者：
Ganesh, Irisappan;Ravikumar, Sambandam;Hong, Soon Ho
为了开发一种高通量筛选系统来筛选产生大量苹果酸的微生物，构建了基于 MalKZ 嵌合 HK 的生物传感器。考虑到大肠杆菌 (E. coli) MalK 与枯草芽孢杆菌 MalK 和大肠杆菌 DcuS 之间的序列相似性，MalK 的假定传感器结构域与 EnvZ 的催化结构域融合。嵌合 MalK/EnvZ TCS 通过同源响应调节因子 OmpR 诱导 ompC 启动子响应细胞外苹果酸。实时定量 PCR 和 GFP 荧光研究表明，随着苹果酸浓度的增加，ompC 基因表达和 GFP 荧光增加。通过使用该策略，可以构建各种基于嵌合TCS的细菌生物传感器，其可用于开发产生生化的重组微生物。
In an attempt to develop a high-throughput screening system for screening microorganisms which produce high amounts of malate, a MalKZ chimeric HK-based biosensor was constructed. Considering the sequence similarity among Escherichia coli (E. coli) MalK with Bacillus subtilis MalK and E. coli DcuS, the putative sensor domain of MalK was fused with the catalytic domain of EnvZ. The chimeric MalK/EnvZ TCS induced the ompC promoter through the cognate response regulator, OmpR, in response to extracellular malate. Real-time quantitative PCR and GFP fluorescence studies showed increased ompC gene expression and GFP fluorescence as malate concentration increased. By using this strategy, various chimeric TCS-based bacteria biosensors can be constructed, which may be used for the development of biochemical-producing recombinant microorganisms.