Comprehensive characterization of recombinant Pichia pastoris using dynamic process conditions and process analytical technology

Abstract

HRP_ FB I HRP_FB II HRP_B II Fig 3. Induction by methanol pulses in a batch culture. Cells were grown using D-glucose as C-source. When the substrate was exhausted, induction of protein expression was carried out by successive pulses of methanol: 1 adaptation pulse with 0,5 % methanol (in gray), successive pulses with 1 % methanol (in orange) and 1 pulse with 2 % methanol (not shown). The on-line calculated carbon dioxide evolution rate (CER) and the oxygen uptake rate (OUR) are shown. These calculations were performed by a metabolic balance analyzer implemented on-line. HRP activity was detected both in the absence and in the presence of D-Ala with slightly better results in the latter case, therefore D-Ala was added to the fed-batch cultures.