Scale-down of continuous protein producing Saccharomyces cerevisiae cultivations using a two-compartment system.
In the biotechnological industry, economic decisions in investment are typically based on laboratory-scale experiments. Scale-down as a tool is therefore of high industrial importance to transfer the processes into larger production scale without loss in performance. In this study, large-scale prolonged continuous cultivations with a heterologous protein producing Saccharomyces cerevisiae strain have been scaled-down to a two-compartment scale-down reactor system. The effects of glucose, pH, and oxygen concentration gradients have been investigated by comparison with corresponding 300 mL standard continuous cultivations. It was found that substrate gradients within a limited range result in increased productivity of the heterologous protein under regulation of glycolytic TPI promoter and delay the decrease of protein and trehalose production during continuous cultivation. Based on these results, it is argued that introduction of variations in substrate concentration can be beneficial for industrial continuous cultivations.
Diabetes Up- and Downstream Development, Novo Nordisk A/S, Bagsvaerd, Denmark.
Naia Risager
NR
Wright
Diabetes Up- and Downstream Development, Novo Nordisk A/S, Bagsvaerd, Denmark.
Nanna Petersen
NP
Rønnest
Dept. of Systems Biology, Technical University of Denmark, Lyngby, Denmark.
Jette
J
Thykaer
D019149:Bioreactors; D001709:Biotechnology; D018929:Cell Culture Techniques / Q000379:methods*; D005656:Fungal Proteins / Q000096:biosynthesis* / Q000737:chemistry; D005947:Glucose / Q000737:chemistry; D006019:Glycolysis; D006863:Hydrogen-Ion Concentration; D010100:Oxygen / Q000737:chemistry; D011401:Promoter Regions, Genetic; D012441:Saccharomyces cerevisiae; D014199:Trehalose / Q000096:biosynthesis / Q000737:chemistry
D005656:Fungal Proteins; D014199:Trehalose; D005947:Glucose; D010100:Oxygen
Continuous cultivation; Heterologous protein production; S. cerevisiae; Scale-down; Two-compartment system