Spatiotemporal Gene Expression by a Genetic Circuit for Chemical Production in Escherichia coli

Abstract

Gene expression in spatiotemporal distribution improves the ability of cells to respond to changing environments. For microbial cell factories in artificial environments, reconstruction of the target compound's biosynthetic pathway in a new spatiotemporal dimension/scale promotes the production of chemicals. Here, a genetic circuit based on the Esa quorum sensing and lac operon was designed to achieve the dynamic temporal gene expression. Meanwhile, the pathway was regulated by an Lcysteine-specific sensor and relocalized to the plasma membrane for further flux enhancement to L-cysteine and toxicity reduction on a spatial scale. Finally, the integrated spatiotemporal regulation circuit for L-cysteine biosynthesis enabled a 14.16 g/L L-cysteine yield in Escherichia coli. Furthermore, this spatiotemporal regulation circuit was also applied in our previously constructed engineered strain for pantothenic acid, methionine, homoserine, and 2-aminobutyric acid production, and the titer increased by 29, 33, 28, and 41%, respectively. These results highlighted the applicability of our spatiotemporal regulation circuit to enhance the performance of microbial cell factories.

Spatiotemporal Regulation and Transport Engineering for Sustainable Production of Geraniol in Candida glycerinogenes

“…The plasma membrane-anchoring strategy initially improved the titer of geraniol, this result suggested that reconstruction of the MVA pathway to a new spatial dimension could facilitate the synthesis of geraniol. However, the biomass of the plasma membrane-anchored strains was significantly lower compared to the control strain CgOYE2Δ (Figure A).…”

“…8,9 Subcellular compartmentalization strategies, a key advantage of eukaryotes over prokaryotes, have been reported to improve protein expression 10 and have been successfully implemented to improve the production of highvalue-added compounds in a variety of organelles, such as mitochondria, 11 peroxisomes, 7,12 endoplasmic reticulum, and lipid droplets. 13,14 Recently, plasma membrane targeting strategies have been used to facilitate the production of valuable chemicals 15 and to address intrapathway competitions. 16 As well, some studies have improved the functional expression and substrate accessibility of membrane-anchored enzymes by relocalizing key rate-limiting enzyme genes to the plasma membrane.…”

“…To address the delivery and accessibility of key intermediates or substrates, spatial optimization of isoprenoid synthesis pathways or key rate-limiting enzymes has been proposed to improve the biosynthesis of target compounds. Subcellular compartmentalization of metabolic pathways would allow higher local concentrations of enzymes, substrates, and intermediates, mitigating toxicity and bypassing the competing pathways, and the control of post-translational levels. , Subcellular compartmentalization strategies, a key advantage of eukaryotes over prokaryotes, have been reported to improve protein expression and have been successfully implemented to improve the production of high-value-added compounds in a variety of organelles, such as mitochondria, peroxisomes, , endoplasmic reticulum, and lipid droplets. , Recently, plasma membrane targeting strategies have been used to facilitate the production of valuable chemicals and to address intrapathway competitions . As well, some studies have improved the functional expression and substrate accessibility of membrane-anchored enzymes by relocalizing key rate-limiting enzyme genes to the plasma membrane. , However, the effect of the plasma membrane-anchoring strategy on the synthesis of monoterpene compounds has not been reported.…”

Synergistic application of atmospheric and room temperature plasma mutagenesis and adaptive laboratory evolution improves the tolerance of Escherichia coli to L‐cysteine

“…[5] In previous studies, elevating the expression of desensitized serine acetyltransferase (SAT) proved to be a successful strategy for increasing the yield of L-cysteine. [5,8] However, many regulatory mechanisms involved in L-cysteine biosynthesis remain unresolved. [9][10][11][12] One of the major issues is the cytotoxicity associated with L-cysteine.…”