Scale-up of a high cell density continuous culture with Pichia pastoris X-33 for the constitutive expression of rh-chitinase.
The feasibility of large-scale production of recombinant human chitinase using a constitutive Pichia pastoris expression system was demonstrated in a 21-L continuous stirred tank reactor. A steady-state recombinant protein concentration of 250 mg/L in the supernatant was sustained for 1 month at a dilution rate of 0.042 h(-1) (equivalent to one volume exchange per day), enabling a volumetric productivity of 144 mg/L d (240 U/L d). The steady-state dry cell weight concentration in this high cell density culture reached 110 g/L. Considering safety and economical aspects, all large-scale cultivations were conducted without molecular oxygen supplementation. Conventional air sparging was used instead. The oxygen demand of the process was determined by off-gas analysis (OUR = 4.8 g O(2) L(-1) h(-1) with k(L)a = 846 h(-1)) and evaluated with regard to further reactor scale-up.
Schilling
B M
BM
Genzyme Corp., One Kendall Square, Cambridge, Massachusetts 02139, USA.
Goodrick
J C
JC
Wan
N C
NC
D002455:Cell Division; D002688:Chitinases / Q000235:genetics* / Q000378:metabolism*; D005285:Fermentation; D005947:Glucose / Q000378:metabolism; D006801:Humans; D007218:Industrial Microbiology / Q000379:methods*; D010100:Oxygen / Q000378:metabolism; D010843:Pichia / Q000235:genetics*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
D011994:Recombinant Proteins; D002688:Chitinases; D005947:Glucose; D010100:Oxygen