Tutorial 3 CLL271

Please DO NOT plagiarize. Severe penalty for non-compliance.

Review: OUR, OTR

1. A genetically modified Pichia pastoris strain is cultured in a bioreactor at 30 ̊C for production

of recombinant human serum albumin. The oxygen requirements for a strain is 97 mmol l-1 h-1
and the critical oxygen requirement is 0.004 mM. The oxygen solubility of culture broth is
estimated to be 7% lower than water due to solute effects.

a) What is the minimum kLa required to sustain the culture with the dissolved oxygen level
above critical if the reactor is sparged with air at approx. 1 atm pressure?
b) What kLa is required if pure O2 is used instead of air?
c) Calculate % decrease in minimum kLa if air is supplied at 1 atm pressure and solute effects
are negligible on dissolved oxygen?

[Solubility of oxygen in water at 30 ̊C under 1 atm air pressure = 8.05 x 10-3 kg m-3. Solubility of
oxygen in water at 30 ̊C under 1 atm oxygen pressure = 3.84 x 10-2 kg m-3]

2. Define oxygen uptake rate and oxygen transfer rate. Consider a unit liquid volume in a
bioreactor with air bubbles in the presence of living cells. Derive the equation for OUR and
OTR? At what condition OUR become equals to OTR?

For an aerobic bioreactor at steady-state with 16g dry wt. cells/L, it was found that the rate of
oxygen uptake by the cells was 850 mg O2 l-1h-1. Assuming that the saturation oxygen
concentration is 5 mg O2 l-1 and a near zero dissolved oxygen concentration, find the volumetric
mass transfer coefficient.

3. A value of kLa = 0.6 /min has been determined for a fermenter at its maximum practical
agitator rotational speed and with air being sparged at 0.5 l gas/reactor volume-min. E. coli with
aq O2 of 10 mmol O2/g-dry wt-hr are to be cultured. The critical dissolved oxygen concentration
is 0.2 mg/l. The solubility of oxygen from air in the fermentation broth is 7.3 mg/l at 30°C.

A. What is the maximum concentration of E. coli that can be sustained in this fermenter under
aerobic conditions?
B. What concentration could be maintained if pure oxygen was used to sparge the reactor (mole
fraction of oxygen in air=0.21)?

4. A strain of E. coli is cultured in a 15 m3 stirred tank fermenter for the production of

heterologous protein. Under the process operating conditions, kLa is 0.2 s-1. The solubility of
oxygen in the broth is 8.05 x 10-3 kg m-3.
a) The specific oxygen uptake rate (OUR) is 15 mmol g-1 h-1. What is the maximum cell
concentration supported by oxygen transfer in the fermenter?
b) The bacteria suffer growth inhibition after reagent X is accidentally added to fermenter broth
just after the start of the cultivation. This results in OUR to 6 mmol g-1 h-1. What maximum cell
density can now be supported by oxygen transfer in the fermenter?