Browsing by Subject "Lysine acetylation"

Now showing 1 - 3 of 3
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    Engineering protein production by rationally choosing a carbon and nitrogen source using E. coli BL21 acetate metabolism knockout strains.
    (Springer Nature, 2019-09-04) Lozano Terol, Gema; Gallego Jara, Julia; Sola Martinez, Rosa Alba; Canovas Diaz, Manuel; De Diego Puente, María Teresa; Bioquímica y Biología Molecular B e Inmunología
    Background: Escherichia coli (E. coli) is a bacteria that is widely employed in many industries for the production of high interest bio-products such as recombinant proteins. Nevertheless, the use of E. coli for recombinant protein production may entail some disadvantages such as acetate overfow. Acetate is accumulated under some culture conditions, involves a decrease in biomass and recombinant protein production, and its metabolism is related to protein lysine acetylation. Thereby, the carbon and nitrogen sources employed are relevant factors in cell host metabolism, and the study of the central metabolism of E. coli and its regulation is essential for optimizing the production of biomass and recombinant proteins. In this study, our aim was to fnd the most favourable conditions for carrying out recombinant protein production in E. coli BL21 using two diferent approaches, namely, manipulation of the culture media composition and the deletion of genes involved in acetate metabolism and Nε-lysine acetylation. Results: We evaluated protein overexpression in E. coli BL21 wt and fve mutant strains involved in acetate metabolism (Δacs, ΔackA and Δpta) and lysine acetylation (ΔpatZ and ΔcobB) grown in minimal medium M9 (inorganic ammonium nitrogen source) and in complex TB7 medium (peptide-based nitrogen source) supplemented with glucose (PTS carbon source) or glycerol (non-PTS carbon source). We observed a dependence of recombinant protein production on acetate metabolism and the carbon and nitrogen source employed. The use of complex medium supplemented with glycerol as a carbon source entails an increase in protein production and an efcient use of resources, since is a sub-product of biodiesel synthesis. Furthermore, the deletion of the ackA gene results in a fvefold increase in protein production with respect to the wt strain and a reduction in acetate accumulation. Conclusion: The results showed that the use of diverse carbon and nitrogen sources and acetate metabolism knockout strains can redirect E. coli carbon fuxes to diferent pathways and afect the fnal yield of the recombinant protein bioprocess. Thereby, we obtained a fvefold increase in protein production and an efcient use of the resources employing the most suitable strain and culture conditions.
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    Regulation of the pyrimidine biosynthetic pathway by lysine acetylation of E. coli OPRTase
    (2022-08-22) Lozano Terol, G.; Gallego Jara, J.; Sola Martinez, R. A.; Ortega Retuerta, A.; Martinez Vivancos, A.; Canovas Diaz, M.; De Diego Puente, María Teresa; Bioquímica y Biología Molecular B e Inmunología
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    The protein acetyltransferase PatZ from Escherichia coli is regulated by autoacetylation-induced oligomerization
    (2015) De Diego Puente, María Teresa; Gallego Jara, Julia; Castaño Cerezo, Sara; Bernal Sanchez, Vicente; Manjón Rubio, Antonio; Cánovas Diaz, Manuel; Fernández Espín, Vanesa; García de la Torre, José; Bioquímica y Biología Molecular B e Inmunología
    Background: PatZ is the main Escherichia coli acetyltransferase and control acetyl-CoA synthetase (Acs) activity. Results: The kinetic and structural PatZ oligomer characteristics were determined. Conclusion: PatZ is a stable tetramer and forms an active octamer by autoacetylation to increase its stability. Significance: PTMs by acetylation have structural and functional roles in the cell.