Abstract
Protein production requires a significant amount of intracellular energy. Eliminating the flagella has been proposed to help Escherichia coli improve protein production by reducing energy consumption. In this study, the gene encoding a subunit of FlhC, a master regulator of flagella assembly, was deleted to reduce the expression of flagella-related genes. FlhC knockout in the ptsG-deleted strain triggered significant growth retardation with increased ATP levels and a higher NADPH/NADP+ ratio. Metabolic flux analysis using a 13C-labeled carbon substrate showed increased fluxes toward the pentose phosphate and tricarboxylic acid cycle pathways in the flhC- and ptsG-deleted strains. Introduction of a high copy number plasmid or overexpression of the recombinant protein in this strain restored growth rate without increasing glucose consumption. These results suggest that the metabolic burden caused by flhC deletion was resolved by recombinant protein production. The recombinant enhanced green fluorescent protein yield per glucose consumption increased 1.81-fold in the flhC mutant strain. Thus, our study demonstrates that high-yield production of the recombinant protein was achieved with reduced flagella formation.
Highlights
When a microorganism is used for industrial purposes, its energy metabolism does not have to be similar to that of natural conditions
There were attempts to redistribute the energy metabolism optimized for industrial condition through metabolic engineering or genome minimization, it can be said that optimizing complex microorganism metabolism for industrial purposes is still at an early stage (Kolisnychenko et al, 2002; Choe et al, 2019)
We demonstrated that the ptsG knockout mutant from the W strain (KCTC1039), Wp, showed improved protein production (Jung et al, 2019)
Summary
When a microorganism is used for industrial purposes, its energy metabolism does not have to be similar to that of natural conditions. In an industrial bioreactor, mixing and agitation facilitate nutrient supplements, so that the energy required for movement to search for nutrients would be significantly reduced (Blais et al, 1997). Much more energy is needed in the metabolism required for recombinant protein or metabolite production. There were attempts to redistribute the energy metabolism optimized for industrial condition through metabolic engineering or genome minimization, it can be said that optimizing complex microorganism metabolism for industrial purposes is still at an early stage (Kolisnychenko et al, 2002; Choe et al, 2019). Flagella contain a thin and long structured filament composed of many proteins (Macnab, 2003)
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