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

• Published in: The Korean journal of chemical engineering Vol. 32; no. 10; pp. 2073 - 2077
• Main Authors: Ravikumar, Sambandam, Ganesh, Irisappan, Maruthamuthu, Murali Kannan, Hong, Soon Ho
• Format: Journal Article
• Language: English
• Published: New York Springer US 01.10.2015; 한국화학공학회
• Subjects: Biotechnology; Catalysis; Chemistry; Chemistry and Materials Science; Industrial Chemistry/Chemical Engineering; Materials Science; 화학공학; Green Fluorescent Protein; Acidic Amino Acids; Chimera Two-component System
• ISSN: 0256-1115; 1975-7220
• DOI: 10.1007/s11814-015-0024-z

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 biochemical-producing recombinant microorganisms can be constructed.