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

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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.

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Funding

This study was financially supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (MSIT) (No. NRF-2015R1A2A2A01004733), Golden Seed Project (213004-04-2-SBA30), Ministry of Agriculture, Ministry of Oceans and Fisheries, Korea Polar Research Institute grant (PE14070), and the Mid-Career Researcher Program through the NRF of Korea funded by the MSIT (NRF-2016R1A2B4008707).

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  1. Sambandam Ravikumar and Yokimiko David contributed equally to this work.

Authors and Affiliations

  1. Department of Biotechnology and Bioengineering, Chonnam National University, 77 Yongbong-ro, Gwangju, 61186, Republic of Korea

    Sambandam Ravikumar & Jong-il Choi

  2. Division of Chemical Engineering and Materials Science, Ewha Womans University, 52 Ewhayeodae-gil, Seodaemun-gu, Seoul, 03760, Republic of Korea

    Yokimiko David & Si Jae Park

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  1. Sambandam Ravikumar
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  2. Yokimiko David
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  4. Jong-il Choi
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Correspondence to Si Jae Park or Jong-il Choi.

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Ravikumar, S., David, Y., Park, S.J. et al. A Chimeric Two-Component Regulatory System-Based Escherichia coli Biosensor Engineered to Detect Glutamate. Appl Biochem Biotechnol 186, 335–349 (2018). https://doi.org/10.1007/s12010-018-2746-y

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