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P. pastoris Fermentation using a BioFlo 110 Benchtop Fermentor

. . . . . .. .200 mL
Glycerol* . . . . . . . . . . . . . . . . . . . . . . . . . .< 400 mL
Methanol* . . . . . . . . . . . . . . . . . . . . . . . . .<2 L
Base (to maintain pH at setpoint)* . . . . .. . .< 250 mL
(*) Added, as required

Pichia trace metals solution, PTM1 consisted of :
Cupric sulfate pentahydrate . . . . . . . . . . . . . . .6 g/L
Sodium iodide . . . . . . . . . . . . . . . . . . . . . . . . .0.08 g/L
Manganese sulfate monohydrate . . . . . . . . .. . .3 g/L
Cobalt chloride (anhydrous) . . . . . . . . . . . .. . .0.5 g/L
Zinc chloride (anhydrous) . . . . . . . . . . . . . .. . .20 g/L
Boric acid . . . . . . . . . . . . . . . . . . . . . . . . . . . .0.02 g/L
Sodium molybdate dihydrate . . . . . . . . . . . . . .0.2 g/L
Ferrous sulfate heptahydrate . . . . . . . . . . . . .. .65 g/L
Biotin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ..0.2 g/L
6N sulfuric acid . . . . . . . . . . . . . . . . . . . . . . . .30 mL/L


Control Setpoints
Setpoints were keyed into the controller prior to inoculation and, except for DO which remained high until culture was introduced, the vessel was allowed to equilibrate prior to inoculation.
Temperature . . . . . . . . . . . . . . . . . . . . . . . 30C
pH . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . 5.0
Dissolved Oxygen . . . . . . . . . . . . . . . . ... . 30%
Agitation . . . . . . . . . . . . . . . . . . . . 300 - 1,200 rpm
(responding automatically to oxygen demand)


Dissolved Oxygen (DO) Control
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