Impact of media components from different suppliers on enterokinase productivity in Pichia pastoris

Abstract

Background The aim of this study was to provide an information about the homogeneity on the level of enterokinase productivity in P. pastoris depending on different suppliers of the media components. Results In previous studies, we performed the optimisation process for the production of enterokinase by improving the fermentation process. Enterokinase is the ideal enzyme for removing fusion partners from target recombinant proteins. In this study, we focused our optimization efforts on the sources of cultivation media components. YPD media components were chosen as variables for these experiments. Several suppliers for particular components were combined and the optimisation procedure was performed in 24-well plates. Peptone had the highest impact on enterokinase production, where the difference between the best and worst results was threefold. The least effect on the production level was recorded for yeast extract with a 1.5 fold difference. The worst combination of media components had a activity of only 0.15 U/ml and the best combination had the activity of 0.88 U/ml, i.e., a 5.87 fold difference. A substantially higher impact on the production level of enterokinase was observed during fermentation in two selected media combinations, where the difference was almost 21-fold. Conclusions Results demonstrated in the present study show that the media components from different suppliers have high impact on enterokinase productivity and also provide the hypothesis that the optimization process should be multidimensional and for achieving best results it is important to perform massive process also in terms of the particular media component supplier .

Docosahexaenoic acid (DHA) production by Aurantiochytrium limacinum using cassava pulp hydrolysate as an alternative low-cost carbon source

“…Yeast extract contains not only nitrogenous substances, but also several vitamins and minerals that are essentials for cell growth (Jacob et al, 2019;Proust et al, 2019;Yin et al, 2019). On the other hand, peptone, which is typically produced by digesting meat or other proteins, contains primarily amino acids and short peptides with less proportions of vitamin and minerals (Krahulec and S ̌afrańek, 2021). This observation agreed well with a report of Gao et al (2013) that yeast extract supported better growth than peptone, but contradict to a study of Hong et al (2011), where peptone was shown to give slightly higher CDM concentration than yeast extract.…”