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A Chimeric Two-Component Regulatory System-Based Escherichia coli Biosensor Engineered to Detect Glutamate

Title
A Chimeric Two-Component Regulatory System-Based Escherichia coli Biosensor Engineered to Detect Glutamate
Authors
Ravikumar, SambandamDavid, YokimikoPark, Si JaeChoi, Jong-il
Ewha Authors
박시재
SCOPUS Author ID
박시재scopus
Issue Date
2018
Journal Title
APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY
ISSN
0273-2289JCR Link

1559-0291JCR Link
Citation
APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY vol. 186, no. 2, pp. 335 - 349
Keywords
DegSEnvZGlutamateEscherichia coliTwo-component system
Publisher
HUMANA PRESS INC
Indexed
SCI; SCIE; SCOPUS WOS scopus
Document Type
Article
Abstract
In this study, we constructed amino acid biosensors that can be used as a high-throughput system to screen microorganisms that produce glutamate. The biosensors are based on two-component regulatory systems (TCRSs) combined with green fluorescent protein (GFP) as a reporter. A chimeric DegS/EnvZ (DegSZ) TCRS was constructed by fusing the N-terminal domain of the sensor kinase DegS from Planococcus sp. PAMC21323 with the catalytic domain of the osmosensor EnvZ from Escherichia coli to control expression of gfp in response to glutamate. gfp was controlled by the ompC promoter through the activated response regulator OmpR-P. The chimeric TCRS-based biosensors showed a 4-fold increase in the fluorescent signal after adding glutamate. A linear correlation was observed between fluorescence intensity and exogenously added glutamate concentration. The chimeric TCRS-based biosensor was used to determine glutamate concentration at the single-cell level by fluorescence-activated cell sorting. Therefore, this biosensor can be used to isolate novel gene products and optimize pathways involved in amino acid production.
DOI
10.1007/s12010-018-2746-y
Appears in Collections:
엘텍공과대학 > 화학신소재공학전공 > Journal papers
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