Metabolic engineering adopts genetic engineering technology to modify, transform and regulate the biochemical metabolic pathway in microbial cells, so as to realize the biosynthesis from cheap raw materials to target chemicals. Compared with chemical synthesis, this biosynthesis method is a more economical, environmental friendly and reliable industrial production method. The biosynthesis from industrial raw materials to target products generally goes through dozens of enzymatic biochemical reactions. Therefore, researchers often regulate the intracellular metabolic network by introducing, knocking out or regulating enzyme genes, which also constitutes a main branch in the field of metabolic engineering.
In recent years, optogenetics has been applied to regulate gene expression of enzyme proteins and optimize intracellular metabolic pathways. Compared with the traditional small molecule chemical signal, the introduction and removal of optical signal is easier, and there is no need to change the medium or reaction conditions.
At present, a large number of products can be obtained by microbial industrial fermentation, including fine biochemical products (amino acids, vitamins, antibiotics, etc.), drug precursors (artemisinin, opioids, etc.), biofuels (ethanol, isobutanol, etc.) and high molecular polymers (polylactic acid, polyhydroxyfatty acid esters, etc.). Therefore, the progress in the field of metabolic engineering has greatly promoted the development of industrial economy.
Creative BioMart has mastered and developed a variety of optical switching systems for metabolic engineering. We have the ability to provide the construction services of cell lines, expression systems and model organisms.
Precise metabolic regulation of engineering cells
The optimization of engineering metabolic pathway requires careful control of the level and time of metabolic enzyme expression. Optogenetic tools are ideal for this precise control because light can be applied and removed immediately without complex medium changes. We provide a light controlled "metabolic valve" based on LOV domain to regulate the gene expression of enzyme protein, so that yeast cells can grow rapidly and produce ethanol under light conditions. The protocol can accurately control the transcription and expression level of enzyme protein in different stages of fermentation, and can be used for the optimization of biofuel fermentation system.
Rapid and reversible control of metabolic flux
In order to maximize the desired product, metabolic engineers usually express the enzyme at a high and constant level. However, permanent pathway activation may have adverse consequences, including competition with essential pathways and accumulation of toxic intermediates. We describe a set of optogenetic tools to trigger the assembly and disassembly of metabolically active enzyme clusters and create a regulated pathway guidance system in microbial metabolic networks.
Switchable metabolic pathway
In the E. coli cell, the switching of two glycolytic and metabolic pathways is realized by photogenetic tools. The intervention of optogenetic tools can optimize the metabolic pathway of microbial intracellular biosynthesis, which has potential application value in the field of metabolic engineering.
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