title: A novel synthetic medium and expression system for subzero growth and recombinant protein production in Pseudoalteromonas haloplanktis TAC125.
abstract: The Antarctic bacterium Pseudoalteromonas haloplanktis TAC125 is a model organism of cold-adapted bacteria. The interest in the study of this psychrophilic bacterium stems from its capability either as a non-conventional system for production of recombinant protein and as a rich source of bioactive compounds. To further explore the biotechnological ability of P. haloplanktis TAC125, we have developed a synthetic medium, containing D-gluconate and L-glutamate (GG), which allows the bacterium to grow even at subzero temperatures. P. haloplanktis TAC125 growing in GG medium at low temperature displays growth kinetic parameters which confirm its spectacular adaptation to cold environment and subzero lifestyle, paving the way to the definition of the underlying molecular strategies. Moreover, in this paper, we report the setup of a finely regulated gene expression system inducible by D-galactose to produce recombinant protein in GG synthetic medium at temperatures as low as -2.5 °C. Thanks to the combination of the novel medium and the new expression system, we obtained for the first time the production of a recombinant protein at subzero temperature, thus providing an innovative strategy for the recombinant production of "difficult" proteins.
authors: Department of Chemical Sciences, University of Naples Federico II, Complesso Universitario Monte Sant'Angelo, via Cinthia, I-80126, Naples, Italy.
authors: F
authors: F
authors: Sannino
authors: GSK Vaccines, Via Fiorentina, 1, 53100, Siena, Italy.
authors: M
authors: M
authors: Giuliani
authors: Department of Chemical Sciences, University of Naples Federico II, Complesso Universitario Monte Sant'Angelo, via Cinthia, I-80126, Naples, Italy.
authors: U
authors: U
authors: Salvatore
authors: Department of Chemical Sciences, University of Naples Federico II, Complesso Universitario Monte Sant'Angelo, via Cinthia, I-80126, Naples, Italy.
authors: G A
authors: GA
authors: Apuzzo
authors: Institute of Protein Biochemistry, National Research Council, Via Pietro Castellino, 111, I-80126, Naples, Italy.
authors: D
authors: D
authors: de Pascale
authors: Laboratory of Microbial and Molecular Evolution, Department of Biology, University of Florence, via Madonna del Piano 6, I-50018, Sesto Fiorentino (Florence), Italy.
authors: R
authors: R
authors: Fani
authors: Laboratory of Microbial and Molecular Evolution, Department of Biology, University of Florence, via Madonna del Piano 6, I-50018, Sesto Fiorentino (Florence), Italy.
authors: M
authors: M
authors: Fondi
authors: Department of Chemical Sciences, University of Naples Federico II, Complesso Universitario Monte Sant'Angelo, via Cinthia, I-80126, Naples, Italy.
authors: G
authors: G
authors: Marino
authors: Department of Chemical Sciences, University of Naples Federico II, Complesso Universitario Monte Sant'Angelo, via Cinthia, I-80126, Naples, Italy.
authors: M L
authors: ML
authors: Tutino
authors: Department of Chemical Sciences, University of Naples Federico II, Complesso Universitario Monte Sant'Angelo, via Cinthia, I-80126, Naples, Italy. erparril@unina.it.
authors: E
authors: http://orcid.org/0000-0002-9002-5409
authors: E
authors: Parrilli
mesh_terms: D003080:Cold Temperature; D003470:Culture Media / Q000737:chemistry*; D015870:Gene Expression; D005818:Genetic Engineering / Q000379:methods; D005822:Genetic Vectors; D031161:Pseudoalteromonas / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism*
chemical_list: D003470:Culture Media; D011994:Recombinant Proteins
keywords: Cold-adapted bacteria; P. haloplanktis TAC125; Recombinant protein production; Subzero growth temperature; Synthetic medium