Process optimization for recombinant protein production in the psychrophilic bacterium Pseudoalteromonas haloplanktis

Maria Giuliani, Ermenegilda Parrilli, Pau Ferrer, Kristin Baumann, Gennaro Marino, Maria Luisa Tutino

    Research output: Contribution to journalArticleResearchpeer-review

    19 Citations (Scopus)

    Abstract

    A new regulated expression system has been developed for recombinant protein production in Pseudoalteromonas haloplanktis, strongly induced by the presence of l-malate in culture medium. In optimized induction conditions, high yields of soluble recombinant model proteins were obtained in small scale expression experiments. However, performances of the new expression system are still not satisfactory for large scale recombinant protein production. To improve the psychrophilic expression system, the influence of medium composition and cultivation operational strategies on biomass concentration, growth rate and protein production have been investigated in this study. A new defined medium, containing branched amino acids (L, I, V) as carbon sources, was developed resulting in over sevenfold increase of reporter enzyme production and threefold of biomass yield with respect to the previously optimized conditions. The new synthetic medium was also tested for the production of a recombinant antibody fragment. In batch cultivation, up to 0.9 mg g dry cell weight -1 of soluble 3H6 Fab were recovered in bacterial periplasm. Moreover, the new optimized medium was tested for P. haloplanktis chemostat cultivation. A continuous process for 3H6 Fab production was developed leading to a volumetric productivity of about 0.2 mg L-1 h-1 of soluble and periplasmic recombinant antibody. © 2011 Elsevier Ltd.
    Original languageEnglish
    Pages (from-to)953-959
    JournalProcess Biochemistry
    Volume46
    DOIs
    Publication statusPublished - 1 Apr 2011

    Keywords

    • 3H6 Fab
    • Antibody fragment
    • Psychrophiles
    • Recombinant protein production
    • Synthetic medium

    Fingerprint Dive into the research topics of 'Process optimization for recombinant protein production in the psychrophilic bacterium Pseudoalteromonas haloplanktis'. Together they form a unique fingerprint.

  • Cite this