Evaluation of Three Methanol Feed Strategies for Recombinant Pichia Pastoris Mut S Fermentation

Abstract

Methylotrophic yeast Pichia pastoris is an ideal host organism for recombinant protein production. However, adequate methanol feed is still a critical point of successful product formation in P. pastoris Mut 5 fermentations. Three methanol feed strategies were tested: an organic vapor sensor, a dissolved oxygen controlled methanol addition and a predetermined model, as well. The organic vapor sensor proved to be unsuitable for methanol concentration measurements when samples were taken from the head space of the bioreactor, but may have the potential to substitute expensive gas analyzers in methanol fed-batch with a suitable selector submerged into the fermentation media. Dissolved oxygen and substrate consumption did not show strict mathematical relation. However, drop of dissolved oxygen tension for periodic methanol addition may be applied for the determination of the substrate concentration in the media. The rate of methanol consumption shows peaks at 0.45 and 0.95% (v/v) substrate concentrations. The rate of the specific methanol consumption of our model organism was determined. Based on the value of 0.023 h`, which is significantly less than suggested by earlier experiments, a successful predetermined methanol feed strategy was set up.

Optimization of the Trichoderma reesei endo-1,4-beta-xylanase production by recombinant Pichia pastoris

“…Therefore, we suggested that the optimal methanol concentration in the media is ranging from 0.5% to 1.0%. Compared with the methanol concentration, there were fewer reports on YNB and initial cell density [30]. Like with the previous study, our results clearly showed that YNB did not have significant effect on recombinant xylanase production.…”

Effect of methanol feeding strategies on production and yield of recombinant mouse endostatin from Pichia pastoris

“…In some cases, metabolism-related method is preferred , and in other cases, nonmetabolism-related method, in which the growth rate can be controlled, is preferred (Zhang et al, 2000a). However, detailed comparisons of various fedbatch strategies for the production of a specific recombinant protein are very limited (Kupcsulik et al, 2001;Minning et al, 2001;Zhang et al, 2000b). Our purpose here is to evaluate several growth strategies for the production of extracellular murine endostatin.…”

The Effect Of Temperature On Recombinant Human Granulocyte Colony Stimulating Factor Production By Pichia pastoris Expression System

“…The rate of methanol conversion determines the available energy and carbon for maintenance, cell growth, product formation and product transport into the media. Overall productivities for cell mass and HSA (human serum albumin) showed similar dependence on methanol concentration as the specific rates (Kupcsulik & Sevella, 2004).…”