TY - JOUR
T1 - Influence of growth temperature on the production of antibody Fab fragments in different microbes: A host comparative analysis
AU - Dragosits, Martin
AU - Frascotti, Gianni
AU - Bernard-Granger, Lise
AU - Vázquez, Felícitas
AU - Giuliani, Maria
AU - Baumann, Kristin
AU - Rodríguez-Carmona, Escarlata
AU - Tokkanen, Jaana
AU - Parrilli, Ermenegilda
AU - Wiebe, Marilyn G.
AU - Kunert, Renate
AU - Maurer, Michael
AU - Gasser, Brigitte
AU - Sauer, Michael
AU - Branduardi, Paola
AU - Pakula, Tiina
AU - Saloheimo, Markku
AU - Penttilä, Merja
AU - Ferrer, Pau
AU - Luisa Tutino, Maria
AU - Villaverde, Antonio
AU - Porro, Danilo
AU - Mattanovich, Diethard
PY - 2011/1/1
Y1 - 2011/1/1
N2 - 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. © 2010 American Institute of Chemical Engineers.
AB - 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. © 2010 American Institute of Chemical Engineers.
KW - Chemostat
KW - Escherichia coli
KW - Pichia pastoris
KW - Pseudoalteromonas haloplanktis
KW - Recombinant protein production
KW - Saccharomyces cerevisiae
KW - Trichoderma reesei
U2 - https://doi.org/10.1002/btpr.524
DO - https://doi.org/10.1002/btpr.524
M3 - Article
VL - 27
SP - 38
EP - 46
JO - Biotechnology Progress
JF - Biotechnology Progress
SN - 8756-7938
ER -