Oxygen transfer as a tool for fine-tuning recombinant protein production by Pichia pastoris under glyceraldehyde-3-phosphate dehydrogenase promoter.

Abstract

Effects of oxygen transfer on recombinant protein production by Pichia pastoris under glyceraldehyde-3-phosphate dehydrogenase promoter were investigated. Recombinant glucose isomerase was chosen as the model protein. Two groups of oxygen transfer strategies were applied, one of which was based on constant oxygen transfer rate where aeration rate was Q O/V=3 and 10 vvm, and agitation rate was N=900min(-1); while the other one was based on constant dissolved oxygen concentrations, C DO=5, 10, 15, 20 and 40% in the fermentation broth, by using predetermined exponential glucose feeding with μ o=0.15h(-1). The highest cell concentration was obtained as 44gL(-1) at t=9h of the glucose fed-batch phase at C DO=20% operation while the highest volumetric and specific enzyme activities were obtained as 4440UL(-1) and 126Ug(-1) cell, respectively at C DO=15% operation. Investigation of specific enzyme activities revealed that keeping C DO at 15% was more advantageous with an expense of relatively higher by-product formation and lower specific cell growth rate. For this strategy, the highest oxygen transfer coefficient and oxygen uptake rate were K L a=0.045s(-1) and OUR=8.91mmolm(-3)s(-1), respectively.