Reconnecting the carbon flux in the metabolic pathway to the optimal state is an important way of efficient biological production. Many studies have been devoted to modification through deletion and overexpression of target genes. Because these modifications sometimes lead to serious growth defects, accumulation of toxic intermediates and imbalance of cofactors, a technique to fine tune the expression level is needed.
Optogenetic switch, a gene regulatory system that controls gene expression level by light, has attracted people's attention. Since we can set the light intensity, wavelength and illumination period arbitrarily, the switch can be used as an on / off switchable tool for metabolic engineering. The reversible switch for target gene induction can be used many times as needed to adjust the appropriate metabolic state. This light induced gene expression system needs to modify proteins by specific wavelength light irradiation.
Creative BioMart provides customers with an optogenetic switching scheme for controlling the flux distribution of glycolysis and methylglyoxal (MGO) pathway. A two-component system CcaS/CcaR is used to operate a optogenetic switch to control transcription in response to green and red illumination.
CcaS and CcaR are optical sensors and their response regulators, respectively. Under green light conditions, CcaS autophosphorylates and transfers its phosphate group to CcaR. Phosphorylated CcaR induces gene expression under the cpcg2 promoter. Under red light, CcaR was dephosphorylated by CcaS, and the gene expression under cpcG2 promoter was inhibited. Using the optimized CcaS/CcaR system in E. coli, the gene expression level under cpcG2 promoter was controlled within a dynamic range of 117 times according to light conditions.
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