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THE UNIVERSITY OF SYDNEY

Faculty of Engineering

School of Chemical and Biomolecular Engineering

CHNG3804/9304 Biochemical Engineering

Semester 2, 2018 Time allowed: two hours

SID:_____________________

Course Code: CHNG3804 OR CHNG9304 (Please Circle)

ANSWER ALL QUESTIONS IN THIS BOOKLET

ANSWERS MUST BE WRITTEN IN THE BOOKLET

Closed book, Non-programmable electronic calculators may be used.

Each Question is worth 20 marks.

Marks for each sub-question are indicated.

Candidates are advised that full marks to problem solutions will only be given when the solution
procedure is clearly explained and all assumptions are clearly stated.
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Question 1 20 marks

If Australian Raw Sugar Exports are~ 4 million tonnes per annum, and molasses production is~
1.2 million tonnes per annum:

a) What fraction of Australia’s Liquid Fuel Consumption of 32 x 109 L could be met via the
fermentation of sugar and molasses? (10 marks)
b) What would be the difference in income between converting all of Australia’s molasses
production to lactic acid compared to ethanol? (10 marks)

You may assume the following:

Molasses is 40% sugar, the density of ethanol is 785 kg/m3.

The Fermentation of sucrose to ethanol is given by

C12H22O11 + H2O  4 C2H5OH + 4 CO2

The Fermentation of sucrose to lactic acid is given by

C12H22O11 + H2O  4C3H6O3

Heats of Combustion

Ethanol 26.7 MJ/kg

Petrol 43 MJ/kg

Ethanol 21.2 MJ/L

Petrol 32.68 MJ/L

Selling Prices

Ethanol $0.50/L

Lactic Acid $1.50/kg

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Answer Question 1 Part A here:

Answer Question 1 Part B here:

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Question 2 20 marks

a) What are the practical consequences of animal cells generally more “shear sensitive” than
bacteria? (2 marks)

b) What type of fermenters are typically used for large scale aerobic processes? (2 marks)

c) What is meant by “Recombinant DNA technology”? What products are typically made
using recombinant DNA Technology? (2 marks)

d) What materials are commonly used for large-scale fermenters? What materials are
commonly used for small-scale fermenters? (2 marks)

e) What methods can be used to sterilise fermentation media? (2 marks)

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f) Why are bubble columns or airlift reactors used in preference to stirred tanks for large
scale fermentations? (2 marks)

g) Why might a temperature profile be used during a fermentation? (2 marks)

h) What is primary, secondary, tertiary and quaternary structure of a protein? Which of these
structures can affect the function of a protein? Give an example of a protein. (2 marks)

h) Give an example of an industrial bioprocess that employs:

a. A batch processes
b. A fed-batch processes (2 marks)
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Question 3 20 Marks

Bacillus subtilis natto is grown to produce Menaquinone 7. The bacteria are grown aerobically in
batch culture with glycerol as the growth-limiting substrate. Cell and substrate concentrations are
measured as a function of culture time; the results are listed below:

Time X (OD600) Glycerol (g/L)

0 1.0 50.0

2 1.4 49.2

4 1.9 48.1

6 2.7 46.5

8 3.8 44.3

10 5.3 41.3

12 7.3 37.2

14 10.1 31.5

16 13.9 23.9

18 18.7 14.0

20 24.2 2.9

22 25.0 1.0

24 24.9 0.0

a) Plot  as a function of substrate concentration. (4 marks)

b) What is max (see page 8 for equation if needed)? (4 marks)
c) What is Km (see page 8 for equation if needed)? (4 marks)
d) If 1 OD600 is approximately equal to 0.5 g/L, what is the yield coefficient Yx/s (see
page 8 for equation if needed)? (4 marks)
e) Comment on whether the Monod equation fits this data well? (4 marks)
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Write your answers to Question 3 below:

b) max = __________

c) km =____________

d) Yxs =____________

e) Suitability of Monod
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Question 4 20 Marks

a. List three methods that can be used to increase the Oxygen Transfer Rate in a bioreactor,
what are the limitations associated with each method. (5 marks)

b. A strain of Azotabacter vinelandii is cultured in a 15m3 stirred tank fermenter. The kla is
0.17s-1 and oxygen solubility in the broth is 8ppm. If the specific rate of oxygen uptake is
12.5mmol g-1h-1 what is the maximum possible cell concentration? (5 marks)
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c. A 150m3 bioreactor is operated at 35oC to produce fungal biomass from glucose. The rate
of oxygen consumption of the culture is 1.5 kgm-3h-1 and the agitator dissipates heat at a
rate of 1 kWm-3. If cooling water at 10oC and a flow of 60 m3h-1 is available from a
nearby river, what is the cooling water outlet temperature? You may assume that the heat
of metabolism is 460kJ/mol of Oxygen consumed (10 marks)
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Question 5 20 Marks

Lactobacillus casei is propagated under essentially anaerobic conditions to provide a starter

culture for manufacture of Swiss cheese. The culture produces lactic acid as a by-product of
energy metabolism. The system has the following characteristics:

Yxs = 0.23 kgkg -1

K s = 0.15 kgm -3

ma x = 0.35 h -1

m s = 0.135 kgkg -l h -l

A stirred fermenter is operated in fed-batch mode at quasi-steady state with a feed flow rate of
4 m3 h-1 and feed substrate concentration of 80 kg m-3. After 6 h, the liquid volume is 40 m3.

a) What was the initial culture volume? (5 marks)

b) What is the concentration of substrate at quasi-steady state? (5 marks)

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c) What is the concentration of cells at quasi-steady state? (5 marks)

d) What mass of cells is produced after 6 h fed-batch operation? (5 marks)

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Information that may be required:

dX μmax S
=μX= X
Batch Growth: dt K m+ S
dS 1
Batch Substrate Uptake: =− μX −mX
dt Y XS

Where X is the biomass concentration, t is the time,  is the specific growth rate, max is the maximum
specific growth rate, S is the substrate concentration, m is the maintenance coefficient, K m is the
substrate concentration at which  0.5max, YXS is the biomass yield from the substrate.

V −V 0
F=
t

where F is the inlet flow rate into the fermenter, V is the volume, V 0 is the initial volume.

F
D=
V
μmax S
D=
K s+ S

where D is the dilution rate, max is the maximum specific growth rate, Ks is the substrate constant and S
is the steady-state substrate concentration in the reactor.

For fed-batch reactor and continuous bioreactors:

dX F μ S μmax S
dt
=μX− X = max −D X
V Km+ S ( ) at (pseudo) steady-state
0=
( K m +S
−D X
)
dS μ D
=D( Si −S )−( +m ) X 0=D(S i−S )−( +m ) X
dt Y XS s at (pseudo) steady-state
Y XS s

At steady-state dS/dt is zero. This condition can be used to determine the critical dilution rate, assuming
that S and ms are almost zero. At steady state we can also assume that S<<S i.

−dX r X 1 1 mS
Y 'XS= = = +
dS T r S Y 'XS Y XS μ
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where ms is the maintenance coefficient, YXS is the true yield coefficient and Y ’XS is the observed yield
coefficient.
2
g ( ρ s −ρw )d
v s=
Stokes’ law (settling velocity): 18 μ 2
1 . 54V where g = 9.8 m/s
t m= 3
Mixing Time (s) in stirred tank reactors: d imp N

H −0.33
t m=11 ( gU g D−2 )
Mixing Time (s) in bubble columns: D

Unit definition: W=kg m2 s-3

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THIS IS THE END OF YOUR EXAMINATION PAPER