BITS Pilani, KK Birla Goa Campus

Midterm test- Semester II (2017-2018)

Course name: Biochemical Engineering Code: CHE F421
Answer all the questions
1. A higher Ks value of Monod's equation means (1 mark)
A. greater affinities to substrate
B. lower affinities to substrate
C. unaffected with the substrate bonding
D. lower dissociation constant value

2. When a culture of fermenting yeast becomes metabolically uncoupled, ethanol (1 mark)

A. becomes a secondary metabolite
B. production becomes growth associated
C. production becomes non-growth associated
D. becomes a growth nutrient

3. During the log/exponential phase of growth in a batch process, µ is always equal to µ max
as (2 marks)
A) Concentration of the growth limiting substrate is much less than the Monod constant
B) Concentration of growth limiting substrate is much greater than the Monod constant
C) Specific growth rate increases exponentially
D) Concentration of the growth limiting substrate is equal to the Monod constant

4. What is High Cell Density fermentation (HCDF)? How it can be achieved, by taking
recombinant protein production as an example? (2 marks)

5. Lipase is being investigated as an additive to laundry detergent for removal of stains from
Fabric. The general reaction is:
Fats  fatty acids + glycerol
The Michaelis constant for pancreatic lipase ig 5 mM. At 60°C lipase is subject to
deactivation with a half-life of 8 min. Fat hydrolysis is carried out in a well-mixed batch
reactor which simulates a top-loading washing machine. The initial fat concentration is
45 gmol m -3. At the beginning of the reaction the rate of hydrolysis is 0.07 mmol 1-1 s -1.
How long does it take for the enzyme to hydrolyze 80% of the fat present? (3 marks)

6. Assume you are developing a inoculum for the production of Rec-Insulin by E.coli in a
stirred tank bioreactor of volume 10000 L using a chemically defined medium with
Glucose as the carbon source. The inoculum train contains 3 inoculum tanks in
sequence, and involves aseptic transfer of 10 % v/v between subsequent stages. The
quality control procedures during inoculum transfer reveals presence of 1 plasmid-free
organism for every 1000 plasmids-bearing bacteria /ml of liquid during the beginning
of inoculum development in tank 1. Assume the specific growth rate of plasmid-free
organism is 1 h-1 and plasmid bearing as 0.5 h-1. Fed-batch process was adopted to grow
 E.coli to high cell density by adding glucose intermittently. After 36 h of fed-batch
feeding, inducer was added to express the hormone. Batch time in each inoculum tank is
24 h and total operation time in final fermentor is 48 h
A) Determine the ratio of productive to non-productive strain at the beginning of
fermentation process. (3 marks)
B) Discuss how the yield of insulin will be affected by the presence non-productive strain
and explain how this problem can be minimized (2 marks)

7. How with the use of electron balance concept, theoretical oxygen demand can be
calculated? (2 marks)

8. An engineer decides to design a special airlift reactor for growing Torula utilis cells. The
reactor relies on evaporative cooling only. Water is evaporated from the broth by
sparging with dry air; water vapor leaving the vessel is then condensed in an external
condenser and returned to the culture. On average, 0.5 kg water is evaporated per day. If
the rate of oxygen uptake by the cells is 140 mmol h-1 and the culture temperature must
be maintained at 30 °C, does evaporation provide adequate cooling for this system? Δhv
water at 30°C = 2430.7 kJ kg-1 (3 marks)

9. What is the name of the phenomenon, whereby yeast produces alcohol during aerobic
growth (despite increased availability of O2) under high glucose concentration? How it
can be minimized during the production of baker’s yeast? (3marks)

10. What is the role of chelator as the medium constituent? (1 mark)

11. Explain the limitation of cell density measurements by optical density method? Prescribe
any other technique which can be used under such circumstances? (2mark)

12. Nicotiana tabacum cells are cultured to high density for production of polysaccharide
gum. The reactor used is a stirred tank that initially contains 100 litres of medium. The
doubling time of the culture is 3 days and the yield of biomass from substrate is 0.5 g g-1.
The concentration of growth-limiting substrate in the medium is 3% (w/v). The reactor is
inoculated with 1.5 g l-1 of cells and operated in batch until the substrate is virtually
exhausted; medium flow is then started at a dilution rate of 0.04 day -1. Fed-batch
operation is carried out for 40 days under quasi-steady-state conditions.
(a) Estimate the batch culture time and the final biomass concentration achieved after the
batch culture period. (2 marks)
(b) What is the final mass of cells in the reactor? (1 mark)
(c) The fermenter is available 250 days per year with a downtime between runs of 24 h.
How much plant cell biomass is produced annually? (2 marks)