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Conflict of Interest: In relation to this article, we declare that there is no conflict of interest.

Publication history: Received September 28, 2014
Accepted January 27, 2015

This is an Open-Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/bync/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Engineering Escherichia coli to sense acidic amino acids by introduction of a chimeric two-component system

Sambandam Ravikumar Irisappan Ganesh¹ Murali Kannan Maruthamuthu¹ Soon Ho Hong^1†

Department of Pharmaceutical Science and Technology, Catholic University of Daegu, 330, Geumrak 1-ri, Hayang-eup, Gyeongsan, Gyeongbuk 712-702, Korea ¹Department of Chemical Engineering, University of Ulsan, 93, Daehak-ro, Nam-gu, Ulsan 680-749, Korea

shhong@ulsan.ac.kr

Korean Journal of Chemical Engineering, October 2015, 32(10), 2073-2077(5), 10.1007/s11814-015-0024-z

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Abstract

In an attempt to create an acidic amino acid-sensing Escherichia coli, a chimeric sensor kinase (SK)-based biosensor was constructed using Pseudomonas putida AauS. AauS is a sensor kinase that ultimately controls expression of the aau gene through its cognate response regulator AauR, and is found only in P. putida KT2440. The AauZ chimera SK was constructed by integration of the sensing domain of AauS with the catalytic domain of EnvZ to control the expression of the ompC gene in response to acidic amino acids. Real-time quantitative PCR and GFP fluorescence studies showed increased ompC gene expression and GFP fluorescence as the concentration of acidic amino acids increased. These data suggest that AauS-based recombinant E. coli can be used as a bacterial biosensor of acidic amino acids. By employing the chimeric SK strategy, various bacteria biosensors for use in the development of biochemicalproducing recombinant microorganisms can be constructed.

Keywords

Acidic Amino Acids Chimera Two-component System Green Fluorescent Protein Escherichia coli

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