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Modelling hCDKL5 Heterologous Expression in Bacteria.
Fondi, Marco; Gonzi, Stefano; Dziurzynski, Mikolaj; Turano, Paola; Ghini, Veronica; Calvanese, Marzia; Colarusso, Andrea; Lauro, Concetta; Parrilli, Ermenegilda; Tutino, Maria Luisa.
Afiliação
  • Fondi M; Department of Biology, University of Florence, Sesto F.no Florence, 50019 Florence, Italy.
  • Gonzi S; Centro Studi Dinamiche Complesse (CSDC), University of Florence, Sesto F.no Florence, 50019 Florence, Italy.
  • Dziurzynski M; Department of Biology, University of Florence, Sesto F.no Florence, 50019 Florence, Italy.
  • Turano P; Department of Environmental Microbiology and Biotechnology, Institute of Microbiology, Faculty of Biology, University of Warsaw, 02-096 Warsaw, Poland.
  • Ghini V; Magnetic Resonance Center (CERM) and Department of Chemistry "Ugo Schiff", University of Florence, via Sacconi 6, Sesto Fiorentino, 50019 Fiorentino, Italy.
  • Calvanese M; Consorzio Interuniversitario Risonanze Magnetiche MetalloProteine (CIRMMP), via Sacconi 6, Sesto Fiorentino, 50019 Fiorentino, Italy.
  • Colarusso A; Magnetic Resonance Center (CERM) and Department of Chemistry "Ugo Schiff", University of Florence, via Sacconi 6, Sesto Fiorentino, 50019 Fiorentino, Italy.
  • Lauro C; Consorzio Interuniversitario Risonanze Magnetiche MetalloProteine (CIRMMP), via Sacconi 6, Sesto Fiorentino, 50019 Fiorentino, Italy.
  • Parrilli E; Dipartimento di Scienze Chimiche, Complesso Universitario Monte Sant'Angelo, 80126 Napoli, Italy.
  • Tutino ML; Dipartimento di Scienze Chimiche, Complesso Universitario Monte Sant'Angelo, 80126 Napoli, Italy.
Metabolites ; 11(8)2021 Jul 28.
Article em En | MEDLINE | ID: mdl-34436432
ABSTRACT
hCDKL5 refers to the human cyclin-dependent kinase like 5 that is primarily expressed in the brain. Mutations in its coding sequence are often causative of hCDKL5 deficiency disorder, a devastating neurodevelopmental disorder currently lacking a cure. The large-scale recombinant production of hCDKL5 is desirable to boost the translation of preclinical therapeutic approaches into the clinic. However, this is hampered by the intrinsically disordered nature of almost two-thirds of the hCDKL5 sequence, making this region more susceptible to proteolytic attack, and the observed toxicity when the enzyme is accumulated in the cytoplasm of eukaryotic host cells. The bacterium Pseudoalteromonas haloplanktis TAC125 (PhTAC125) is the only prokaryotic host in which the full-length production of hCDKL5 has been demonstrated. To date, a system-level understanding of the metabolic burden imposed by hCDKL5 production is missing, although it would be crucial for upscaling of the production process. Here, we combined experimental data on protein production and nutrients assimilation with metabolic modelling to infer the global consequences of hCDKL5 production in PhTAC125 and to identify potential overproduction targets. Our analyses showed a remarkable accuracy of the model in simulating the recombinant strain phenotype and also identified priority targets for optimised protein production.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Idioma: En Revista: Metabolites Ano de publicação: 2021 Tipo de documento: Article

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Idioma: En Revista: Metabolites Ano de publicação: 2021 Tipo de documento: Article