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An Engineered Rare Codon Device for Optimization of Metabolic Pathways
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Journal Article
An Engineered Rare Codon Device for Optimization of Metabolic Pathways
2016
Overview
Rare codons generally arrest translation due to rarity of their cognate tRNAs. This property of rare codons can be utilized to regulate protein expression. In this study, a linear relationship was found between expression levels of genes and copy numbers of rare codons inserted within them. Based on this discovery, we constructed a molecular device in
Escherichia coli
using the rare codon AGG, its cognate tRNA (tRNA
Arg
(CCU)), modified tRNA
Asp
(GUC → CCU), and truncated aspartyl-tRNA synthetase (TDRS) to switch the expression of reporter genes on or off as well as to precisely regulate their expression to various intermediate levels. To underscore the applicability of our work, we used the rare codon device to alter the expression levels of four genes of the fatty acid synthesis II (FASII) pathway (i.e. fabZ, fabG, fabI, and tesA’) in
E. coli
to optimize steady-state kinetics, which produced nearly two-fold increase in fatty acid yield. Thus, the proposed method has potential applications in regulating target protein expression at desired levels and optimizing metabolic pathways by precisely tuning
in vivo
molar ratio of relevant enzymes.
Publisher
Nature Publishing Group UK,Nature Publishing Group
Subject
/ Aspartate-tRNA Ligase - genetics
/ Aspartate-tRNA Ligase - metabolism
/ Codon
/ Codons
/ E coli
/ Enzymes
/ Escherichia coli - metabolism
/ Escherichia coli Proteins - genetics
/ Escherichia coli Proteins - metabolism
/ Fatty Acid Synthase, Type II - genetics
/ Fatty Acid Synthase, Type II - metabolism
/ Humanities and Social Sciences
/ Metabolic Networks and Pathways
/ Science
/ tRNA
