title: Influence of growth temperature on the production of antibody Fab fragments in different microbes: a host comparative analysis.
abstract: Microorganisms encounter diverse stress conditions in their native habitats but also during fermentation processes, which have an impact on industrial process performance. These environmental stresses and the physiological reactions they trigger, including changes in the protein folding/secretion machinery, are highly interrelated. Thus, the investigation of environmental factors, which influence protein expression and secretion is still of great importance. Among all the possible stresses, temperature appears particularly important for bioreactor cultivation of recombinant hosts, as reductions of growth temperature have been reported to increase recombinant protein production in various host organisms. Therefore, the impact of temperature on the secretion of proteins with therapeutic interest, exemplified by a model antibody Fab fragment, was analyzed in five different microbial protein production hosts growing under steady-state conditions in carbon-limited chemostat cultivations. Secretory expression of the heterodimeric antibody Fab fragment was successful in all five microbial host systems, namely Saccharomyces cerevisiae, Pichia pastoris, Trichoderma reesei, Escherichia coli and Pseudoalteromonas haloplanktis. In this comparative analysis we show that a reduction of cultivation temperature during growth at constant growth rate had a positive effect on Fab 3H6 production in three of four analyzed microorganisms, indicating common physiological responses, which favor recombinant protein production in prokaryotic as well as eukaryotic microbes.
authors.affiliation: Dept. of Biotechnology, University of Natural Resources and Life Sciences, Vienna, Austria.
authors.forename: Martin
authors.initials: M
authors.lastname: Dragosits
authors.forename: Gianni
authors.initials: G
authors.lastname: Frascotti
authors.forename: Lise
authors.initials: L
authors.lastname: Bernard-Granger
authors.forename: Felícitas
authors.initials: F
authors.lastname: Vázquez
authors.forename: Maria
authors.initials: M
authors.lastname: Giuliani
authors.forename: Kristin
authors.initials: K
authors.lastname: Baumann
authors.forename: Escarlata
authors.initials: E
authors.lastname: Rodríguez-Carmona
authors.forename: Jaana
authors.initials: J
authors.lastname: Tokkanen
authors.forename: Ermenegilda
authors.initials: E
authors.lastname: Parrilli
authors.forename: Marilyn G
authors.initials: MG
authors.lastname: Wiebe
authors.forename: Renate
authors.initials: R
authors.lastname: Kunert
authors.forename: Michael
authors.initials: M
authors.lastname: Maurer
authors.forename: Brigitte
authors.initials: B
authors.lastname: Gasser
authors.forename: Michael
authors.initials: M
authors.lastname: Sauer
authors.forename: Paola
authors.initials: P
authors.lastname: Branduardi
authors.forename: Tiina
authors.initials: T
authors.lastname: Pakula
authors.forename: Markku
authors.initials: M
authors.lastname: Saloheimo
authors.forename: Merja
authors.initials: M
authors.lastname: Penttilä
authors.forename: Pau
authors.initials: P
authors.lastname: Ferrer
authors.forename: Maria
authors.initials: M
authors.lastname: Luisa Tutino
authors.forename: Antonio
authors.initials: A
authors.lastname: Villaverde
authors.forename: Danilo
authors.initials: D
authors.lastname: Porro
authors.forename: Diethard
authors.initials: D
authors.lastname: Mattanovich
mesh_terms: D001419:Bacteria / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D007140:Immunoglobulin Fab Fragments / Q000096:biosynthesis*; D013045:Species Specificity; D013696:Temperature; D015003:Yeasts / Q000378:metabolism*
chemical_list: D007140:Immunoglobulin Fab Fragments