PROBLEMS IN BIOTECH

1. A continuous process is set for the treatment of wastewater. Each day, 105 kg of cellulose and
103 kg bacteria enter in the feed stream, while 104 kg cellulose and 1.5 x 104 kg bacteria leave in
the effluent. The rate of cellulose digestion by the bacteria is 7x104 kg/day. The rate of bacterial
growth is 2x104 kg/day; the rate of cell death by lysis is 5x102 kg/day
Which of the following is true.
a. There is accumulation of both bacteria and cellulose
b. There is depletion of bacteria
c. There is accumulation of cellulose
d. The system is at steady state.

2. Depending on culture conditions, glucose can be catabolised by yeast to produce ethanol and
carbon dioxide, or can be diverted into other biosynthetic reactions. An innoculum of yeast is
added to a solution containing 10 g/L glucose. After some time, only 1 g/L glucose remains while
the concentration of ethanol is 3.2 g/l Determine the fractional conversion of glucose to ethanol.
a. 100% b. 75% c. 70% d. 65%

3. The over-all reaction for microbial conversion of glucose to L-glutamic acid is:
C 6 H 12 O6  NH 3  1.5O2    C 5 H 9 NO4  CO2  3H 2 O
What mass of oxygen is required to produce 15 grams of glutamic acid?
a. 1.86 g b. 3.89 g c. 4.90 g d. 14.9 g
4. A fermentation slurry containing Streptomyces kanamyceticus cells is filtered using a continuous
rotary vacuum filter. Slurry is fed to the filter at a rate of 120 kg/h; 1 kg slurry contains 60 g cell
solids> To improve filtration rates, particles of diatomaceous earth filter aid are added at a rate
of 10 kg/h. The concentration of kanamycin in the slurry is 0.05% by weight. Liquid filtrate is
collected at a rate of 112 kg/h; the concentration of kanamycin in the filtrate is 0.045% (w/w).
Filter cake containing cells and filter aid is removed continuously from the filter cloth. What
percentage water is the filter cake?
a. 1.80% b. 4.39% c. 4.90% d. 14.9%

5. If the concentration of kanamycin dissolved in the liquid within the filter cake is the same as that
in the filtrate, how much kanamycin is absorbed per kg of filter aid?
a. 0.000964 b. 0.0043 c.0.004% d. 0.000149

6. Corn-steep liquor contains 2.5% invert sugars and 50% water; the rest can be considered solids.
Beet molasses contains 50% sucrose, 1% invert sugars, and 18% water; the remainder is solids. A
mixing tank contains 125 kg corn-steep liquor and 45 kg molasses; water is being added to
produce a diluted sugar mixture containing 2% (w/w) Total Sugar as Invert.
How much water is required? (Express sucrose as invert sugar)
a. 1806 kg b. 3,890 k g c.1,170k g d. 1490 k g

7. Acetobacter aceti bacteria convert ethanol into acetic acid under aerobic conditions. A
continuous fermentation process for vinegar production is proposed using non-growing A. Aceti
cells immobilized on the surface of gelatine beads. Air is pumped into the fermenter at a rate of
of 200 gmol/h. The production target is 2 kg/h acetic acid and the maximum acetic acid
concentration tolerated by the cells is 12 % w/w. What is the minimum amount of ethanol
required?
a. 1.86 kg b. 3.80 kg c.1.53kg d. 1.40 kg

1
8. What is the water required for dilution?
a. 18.06 kg b. 38.90 kg c.14. 07 kg d. 24.90 kg

9. What is the percent oxygen in the fermenter off-gas?

a. 18.06 % b. 3.89 % c. 5.16% d. 14.90 %

10. Xanthan gum is produced using Xanthomonas campestris in batch culture. Laboratory
experiments have shown that for each gram of glucose utilized by the bacteria, 0.23 g oxygen and
0.01 g ammonia are consumed, while 0.75 g gum, 0.09 g cells, 0.27 g gaseous CO2, and 0.13 g H2O
are formed. Other components of the system such as phosphate can be neglected. Medium
containing glucose and ammonia dissolved in 20,000 liters of water is pumped into a stirred
fermenter and inoculated with X.campestris. Air is sparged into the fermenter; the total amount
of off-gas recovered during the entire batch culture is 1250 kg. Because xanthan gum solutions
have high viscosity and are difficult to handle, the final gum concentration should not be allowed
to exceed 3.5 wt %. How much glucose is required?
1 g glu cos e  0.23 g O2  0.01 g NH 3  0.75 g gum  0.09 g cells  0.27 g CO2  0.13 g H 2 O
a. 980 kg b. 1250 kg c. 1500 kg 1650 kg

Conditions:
1. No leaks
2. Inlet air and off-gas are dry
3. Conversion of glucose and NH3 is 100% complete
4. Solubility of CO2 and N2 in the liquid phase is negligible
5. CO2 leaves in off-gas

11. How much ammonia is required?

a. 9.80 kg b. 125.0 kg c. 15.00 kg 16.50 kg

12. What percentage excess air is provided?

a. 25% b. 12 % c. 15% d. 18 %

13. The chemical reaction equation for respiration of glucose is:

C6 H12O6  6O2  6CO2  6H 2O
Candida utilis cells convert glucose to CO2 and H2O during growth. The cell composition is
CH1.84O0.55N0.2 plus 5% ash. The yield of biomass from substrate is 0.5 g/g. Ammonia is used as the
nitrogen source. What is the oxygen demand with growth compared to that without?
a. 2.52 b. 1.2 c. 1.5% d. 1.8 %

14. Production of single-protein from hexadecane is described by the following reaction equation:
C16 H 34  a O2  b NH 3  c CH1.66O0.27 N 0.20  d CO2  e H 2 O
Where: CH1.66O0.27N0.20 represents the biomass.

If the respiratory quotient, RQ =0.43, determine the stoichiometric coefficients.

2
15. Saccharomyces cerevisae is grown anaerobically in continuous culture at 30oC. Glucose is used
as the carbon source; ammonia is the nitrogen source. A mixture of glycerol and ethanol is
produced. At steady state, the net mass flows to and from the reactor are as follows.
Data given as follows:

Mass flow, Heat of Combustion Molecular Wt,

kg/h (Δhc) Kg/kgmol
Glucose in 36 -2,805.0 kJ/gmol 180
NH3 in 0.4 -382.6 kJ/gmol 17
Cells out 2.81 -21.2 kJ/g
Glycerol out 7.94 -1,655.4 kJ/gmol 92
Ethanol out 11.9 1,366.8 kJ/gmol 46
CO2 out 13.6 0
H2O out 0.15 0

Reaction: glucose  NH 3  glycerol  ethanol  CO2  H 2 O

Estimate the cooling requirements, kJ/h
a. 13,920 b. 12,4000 c. 15,000 d. 18,000

16. Citric acid is manufactured using submerged culture Aspergillus niger in a batch reactor operated
at 30oC. Over a period of 2 days, 2,500 kg glucose and 860 kg oxygen are consumed to produce
1,500 kg citric acid, 500 kg biomass, and other products. Ammonia is used as the nitrogen source.
Power input to the system by mechanical agitation of the broth is about 15 kw; approximately
100 kg of water is evaporated over the culture period.
Data:
Δhv=λv water at 30oC= 2,430.7 kJ/kg;
H rxn for fully aero bic reaction  460 kJ/gmol oxygen consumed
glucose  O2  NH3  biomass  citric acid  CO2  H 2O
Estimate the cooling requirements.
a. 1.39 x107kJ b. 1.24 x107kJ c. 1.5 x107kJ d.1.47x107kJ