UNIVERSITY COLLEGE LONDON

EXAMINATION FOR INTERNAL STUDENTS

MODULE CODE

ASSESSMENT
PATTERN

BENGM002

BENGM002A

Design and Control of Biochemical Reactors

(Masters Level)

MODULE NAME

DATE

18-May-11

TIME

10:00

TIME ALLOWED

3 Hours 0 Minutes

2010/11-BENGM002A-001-EXAM-3
2010 University College London

TURN OVER

Answer FOUR QUESTIONS. Any extra answers will not be marked. Marks
distributed as shown [ J
1.
a)

A wild-type strain of E. coli is to be cultured in a 2000L working

volume, pilot scale fermenter. The batch fermentation process uses
glucose as a carbon and energy source and NH4OH as the primary
nitrogen source. Laboratory scale studies have indicated that the yield of

biomass on oxygen is around 1.5 goxygen gdow1 and the elemental

composition of the cells produced is C1-11.7900.5NO2 (comprising 7.1%
w/w ash). If the target biomass concentration is 16.5 gdew L-1 calculate
the mass of each of the nutrients required. Clearly state any assumptions
made.
b)

[21]

If the wild-type E. coli is subsequently engineered to enable the

constitutive over-expression of a heterologous protein explain how the
stoichiometric mass balance would be altered.

[4]

2.
a)

You are required to design a 500 L pilot scale fermenter for the batch
culture of a recombinant E. coli strain. Two existing Rushton turbine
impellers are to be reused in order to reduce capital costs. These each
have six flat blades with a width of 3 cm. Specify the dimensions of the
fermenter and the size and location of the internal components. Clearly
state and justify any assumptions made.

b)

[13]

The correlations below are commonly used to estimate the gassed power
requirement, Pg, in stirred-tank fermenters:
0.45

Pg = 0.72[

Q0.56

-025

P
Pg=

01(Ary
2)

-112

Na

(gTpV2 /3J

where Pug, is the ungassed power (W), Q is the volumetric gas flow rate
(m3 5-1), V is the fermenter working volume (m3), N is the impeller
rotational speed (II), di is the impeller diameter and Wi is the width of

the impeller blade (m). Use both correlations to predict Pg/V and
comment on likely reasons for any differences obtained.

Normal operating conditions for this vessel would be: impeller speed =
500 rpm, aeration rate = 0.75 vvm. Lab scale studies have indicated that
the physical properties of the broth are as follows: density = 1020 kg m[12]

, viscosity = 0.02 Ns m-2.

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3.
a)

A fermentation process for a technical enzyme needs to be transferred

from the pilot plant to production scale using a constant kLa as scale-up
criterion.

i)

Explain briefly why a constant kLa is often used for scale-up and
describe the factors that have an influence on the magnitude of kLa.

ii)

[8]

Derive the agitation rate for a 25 m3 (total volume) production reactor

with 80% fill volume using a kLa value of 450 h-land given that the
maximum air flow rate is 0.5 vvm. The vessel has an aspect ratio of 3:1
and is equipped with 3 Rushton turbines (Power number/turbine = 5.7).

The tank to impeller diameter ratio is 3:1. The broth has a density of
1050 kg al3 and a viscosity of 0.03 Ns 1112.

[12]

kLa correlation: kLa = 0.026 (pgN)0.5 (vs)0A

b)

Discuss briefly the implications of fermentation scale-up using constant

tip speed on cell growth of filamentous organisms and oxygen transfer.

[5]

4.
a)

A fermentation process is to be carried out in chemostat culture using a

single growth limiting substrate.

i)

Derive an expression for a well-mixed, single stage chemostat showing

that the specific growth growth rate is equal to the dilution rate at steady
state.

ii)

[7]

Calculate the steady-state growth limiting substrate concentration given

that the input rate of substrate is 500 mT lfland the chemostat has a
working volume of 3 L. It is assumed that cell growth can be described

by the Monad equation and the organism has a substrate affinity

constant of 6 mg L" and a maximum specific growth rate of 0.8 ICI.

[5]

b)

Industrial fermentations are often run in fed-batch mode.

i)

Describe the key features of a fed-batch operation and give examples for
its application.

[6]

Briefly outline possible modes of operation using feedback control.

[7]

ii)

CONTINUED

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5.

It is crucial to control the pH during a fermentation processes to

optimise cell growth. A pH control system has been used to achieve the
real-time pH control in an E coli fermentation for antibody fragment
production. The block diagram of the pH control system is illustrated in
Figure 1 where a pH sensor, a PI controller, and a pump as the activators
for adding base are used. The desirable pH is 6.95. The accepted error is
0.25.

Feeciforward path

Contrail

Measured
output

i-411.

ctuator

Output
Process

Measurement
device
Feedback path

a)

Describe briefly the role of the pH sensor, the PI controller and the
pump. If the pH in the fermentation broth is 6.5 at a particular time, how
will the system respond?

c)

[10]

Due to the noise in the data from the sensor, a first order filter is used:

fiu(t) = (1- a)y(t) + ay flit(t At)

where y(t) represents the measured temperature in the fermenter and
yifit(t) is the signal after the noise has been removed from y(t), t is time,

At is the time interval, and a is a design parameter. Explain how the

c)

filter works and where the filter is allocated.

[5]

Describe the general rules for Pipe & Instrument Drawing (or P&ID) of
a control system and draw the P&ID for the above pH control system.

[10]

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6:

The natural enzymes El and E2 have recently been isolated from a plant

and expressed from a plasmid in E.coli.

El and E2 catalyse the

following bioconversions:

OH

R-NH2
H 001

El

E2

a)

What classes of enzymes do El and E2 each belong to?

[4]

b)

What choice of biocatalyst form is best suited to the use of enzyme El?
Give reasons for your answer.

[5]

c)

A Biochemical Engineer wishes to produce C shown above, at large

scale, starting from compound A. Briefly list some of the problems that
might be encountered in relation to the substrate (A) used, product (C)

made, and activity of the enzymes under the large-scale process

conditions.

[8]

Briefly describe a directed evolution experiment, to improve the stability

of the E2 enzyme in the presence of an organic solvent. Include the
methods that could be used to alter the amino-acid sequence of E2, and a
short description of the overall directed evolution process.

[8]

END OF PAPER

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