Coenzyme Q production by metabolic engineered Escherichia coli strains in defined medium
2019
Martínez, Irene | Zelada, Patricio | Guevara, Felipe | Andler, Rodrigo | Urtuvia, Viviana | Pacheco-Leyva, Ivette | Díaz-Barrera, Alvaro
Coenzyme Q (CoQ) plays an important role as an electron transporter in the respiratory chain. It is formed from a benzoquinone ring and an isoprenoid chain of a specific length depending on the organism. We constructed an engineered Escherichia coli strain (menF) unable to produce demethylmenaquinone and menaquinone, compounds that compete for both chorismate, precursor of the benzoquinone ring, and the isoprenoid chain involved in CoQ biosynthesis. In addition, a mutant strain (entC) unable to produce enterobactin, high-affinity siderophore, synthesized from chorismate, and a double mutant (entC–menF) were constructed. The use of glucose or glycerol as carbon sources was also evaluated for the production of CoQ₈ in these strains. The double mutant (entC–menF) showed 18% increase in CoQ₈-specific content compared to the control strain; however, the single-mutant strains did not show statistically significant differences in CoQ₈-specific content respect to the control, in glucose medium in bioreactor experiments. Glycerol was significantly superior compared to glucose for the production of CoQ₈ in E. coli, where the CoQ₈-specific content increased 126% and 53% in the control and double-mutant strain, respectively. The expression of genes related to CoQ₈ biosynthesis is reported, where the entC–menF double-mutant strain showed a significant increase in the expression of CoQ₈ biosynthesis-related genes when glycerol was used as sole carbon source. The control strain did not show gene expression difference between both carbon sources, indicating a possible regulation at a different level.
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