Assignment 3
1. Serratia marcescens bacteria are used for the production of threonine. The maximum specific
oxygen uptake rate of S. marcescens in batch culture is 5 mmol O2 g 1 h-1. It is planned to
operate the fermenter to achieve a maximum cell density of 40 g l -1 . At the fermentation
temperature and pressure, the solubility of oxygen in the culture liquid is 8 * 10 -3 kg m -3. At a
particular stirrer speed, kLa is 0.15 s-1. Under these conditions, will the rate of cell metabolism
be limited by mass transfer or depend solely on metabolic kinetics?
2. A genetically engineered strain of yeast is cultured in a bioreactor at 30ºC for production of
heterologous protein. The oxygen requirement is 80 mmol l-1h-1; the critical oxygen
concentration is 0.004 mM. The solubility of oxygen in the fermentation broth is estimated to be
10% lower than in water due to solute effects. What is the minimum mass transfer coefficient
necessary to sustain this culture with dissolved oxygen levels above critical if the reactor is
sparged with air at approximately 1 atm pressure? (at 30ºC and 1 atm air pressure, oxygen
solubility is 8.05 * 10-3 kg m-3)
3. A strain of Azotobacter vinelandii is cultured in a 15-m3 stirred fermenter for alginate produc-
tion. Under current operating conditions, kLa is 0.17s-1. The solubility of oxygen in the broth
isapproximately 8 X10-3 kg m-3.
(a) The specific rate of oxygen uptake is 12.5 mmol g-1h-1. What is the maximum cell
concentration supported by oxygen transfer in the fermenter?
(b) The bacteria suffer growth inhibition after copper sulphate is accidentally added to the
fermentation broth just after the start of the culture. This causes a reduction in the
oxygen uptake rate to 3 mmol g -1h-1. What maximum cell concentration can now be
supported by oxygen transfer in the fermenter?
4. A 20-litre stirred fermenter containing Bacillus thuringiensis is used to produce a microbial
insecticide. The oxygen balance method is applied to determine k La. The fermenter operating
pressure is 1.5 atm and the culture temperature is 30ºC. The oxygen tension in the broth is
measured as 82% using a probe calibrated to 100% in situ using water and air at 30ºC and 1.5
atm. The solubility of oxygen in the culture fluid is the same as in water (at 30ºC and 1 atm air
pressure, oxygen solubility is 8.05 * 10-3 kg m-3). Air (containing 20.99% oxygen) is sparged into
the vessel; the inlet gas flow rate measured outside the fermenter at 1 atm pressure and 22ºC is
0.23 l s-1. The exit gas from the fermenter contains 20.1% oxygen and has a flow rate of 8.9 l
min-1. (R = 0.082 l atm K-1 gmol-1)
(a) Calculate the volumetric rate of oxygen uptake by the culture.
(b) What is the value of kLa (assuming composition of gas phase to be equal to its exit
concentration in the fermenter)?
5. A stirred fermenter is used to culture haematopoietic cells isolated from umbilical cord blood.
The liquid volume is 15 litres. The simple dynamic method is used to determine k La . The air
flow is shut off for a few minutes and the dissolved oxygen level drops; the air supply is then
reconnected at a flow rate of 0.25 l s -1. The following results are obtained at a stirrer speed of
50 rpm.
When steady state is established, the dissolved oxygen tension is 78% air saturation. Estimate
kLa neglecting errors associated with the electrode response and gas-phase dynamics.
