Rationale-based selection of optimal operating strategies and gene dosage impact on recombinant protein production in Komagataella phaffii (Pichia pastoris).

摘要: Its features as a microbial and eukaryotic organism have turned Komagataella phaffii (Pichia pastoris) into an emerging cell factory for recombinant protein production (RPP). As key step of the bioprocess development, this work aimed to demonstrate importance tailor designing cultivation strategy according kinetics factory. For purpose, K. phaffii clones constitutively expressing (PGAP ) Candida rugosa lipase 1 (Crl1) with different gene dosage were used models in continuous fed-batch cultures. Production parameters much greater multicopy clone (MCC) than single-copy (SCC). Regarding kinetics, specific product generation rate (qP increased linearly increasing growth (µ) SCC; by contrast, qP exhibited saturation MCC. A transcriptional analysis chemostat cultures suggested presence eventual post-transcriptional bottlenecks After strain characterization, order fulfil overall development bioprocess, performance both was also evaluated mode. Strikingly, optimal strategies determined due kinetic patterns observed trade-off titre, yields productivity. The combined effect adequate µ enables rational process view optimize RPP bioprocesses.