Batch Culture

Q1. A strain of E.coli is grown in batch culture which is started by inoculating 12g
cells into a 100L bubble column fermenter containing 10gl-1 Gucose.

μm = 0.9 hr-1 ; YX/S = 0.575 g glc/g cells

X0 = 12/100 = 3/25 gl-1

S0 = 10 gl-1

a) Estimate the time required to reach stationary culture

b) What will be the final cell density if 70% of substrate is consumed? Assume
there is no product formation and negligible maintenance

Fed-Batch Culture

Q2. In recombinant E.coli cultivation at the start of the fed-batch, the volume of
the reactor is 1 L and the biomass concentration is 1 gl-1. The yield coefficient is
0.5 and substrate concentration in feed is 100 gl-1. It was decided to maintain a
growth rate of 0.3 hr-1. What will be the feed rate at the start of fed-batch? It
was decided to change the feed rate every 20 minutes manually.

Q3. In a fed-batch culture operating intermittent addition of glucose solution

values of the following parameters were give at time t=2 hrs. When the system
is at quasi steady state,

V=1000ml

F =200ml/hr

S0=100gl-1

μm = 0.3 hr-1

YX/S = 0.5 g glc/g cells

X0t = 30 g

Find 1) V0

2) Determine concentration and total amount of biomass in the vessel at t=2 hrs

3) If qp is 0.2 gg-1 and V0= 0, determine concentration of product in at t= 2hrs

Q4. Plant cells are cultured in fed-batch cultivation. The reactor used is a stirred
tank reactor containing initially 100 L medium. The maximum specific growth
rate of the culture is 0.18 day-1. The yield of biomass from substrate is 0.5 gg-1.
The concentration of growth limiting substrate in the medium is 3% W/V. The
reactor is inoculated with 1.5 gl-1 cells and operated in batch mode until the
substrate is exhausted. Medium flow is started at the rate of 4 L per day. Fed-
batch operation occurs under quasi steady state.
a) Estimate the batch phase cultivation time and biomass concentration

b) Fed- batch operation is carried out for 40 days. What is the final mass of cells
in reactor?

c) The reactor is available 275 days per year with the downtime between runs of
24 hours. How much plant cell mass is produced annually?

Q5. A lactobacillus culture has the following characteristics

YX/S= 0.23g/g

KS= 0.15 kg/m3

μm= 0.35 hr-1

ms= 0.135

a stirred fermenter is operated in fed-batch mode at quasi steady state with a

feed flow rate of 4m3/hr and feed substrate concentration of 80 kg/m3 = S0. After
6 hours, the liquid volume is 40m3.

a) What was the initial culture volume?

b) What is the concentration of substrate at quasi steady state?

c) What is the concentration of cells at quasi steady state?

d) What mass of cells is produced after 6 hours of fed batch operation?

Cell-Recycle Cultivation

Q6. Consider a 1000L CSTR in which biomass is produced with glucose as the
substrate. The microbial system follows a Monod relationship with

μmax= 0.4 hr-1

YX/S= 0.5 g/g

KS= 1.5 g/l

If normal operation is with the sterile feed containing 10 g/l glucose at the rate of
100 L per hour

a) What is the specific biomass production rate at steady state

b) If recycle is used with a recycle stream of 10 L/hr and a recycle biomass

concentration 5 times as large as that in the reactor exit. What would be the new
biomass production rate?

Q7. A strain of yeast is being cultivated in a 30L litre CSTR with cell recycling
system. The cell settler was designed so that the cell concentration of its outlet
stream is 30% of that of the inlet stream, whereas the substrate concentration is
the same. The growth rate of the cells can be represented by Monod kinetics with
parameters:

μmax= 0.3 hr-1

YX/S= 0.25 g/g

KS= 0.05

The inlet substrate concentration is 100 g/l and the flow rate is 20 L/hr. The feed
stream is sterile. Calculate the steady state substrate and cell concentration in
the fermenter.

Activated Sludge Reactor - Settling Tank

Q8* don’t know the exact question, but these were the values given in the
question

μmax= 0.2 hr-1

YX/S= 0.5 mg/mg

KS= 50 mg/l

Kd= 0.005 hr-1

V = 3200 m3

α= 0.4

Xe= 0

Θc= 120 hr

S0=800 mg/L

F= 400 m3/hr

Find S

Q9. The BOD value of waste water feed stream is S0= 300 mg/L and the effluent
is desired to be 30mg/L. The feed flow rate is F = 2* 107 L/day. For the recycle
ratio of 0.5 and steady state biomass concentration of 5 g/L, Calculate the
following

a) Required reactor volume

b) Biomass concentration in the recycle

c) Solid residence time Θc

d) Hydraulic residence time ΘH

Given: μm= 1.5 day-1

YX/S= 0.5 g/g

KS= 400 mg/l

Plug Flow, Immobilised System

Q10. Immobilized lactase is used to hydrolyse lactose in dairy wastes to glucose

and galactose. Enzyme is immobilized in resin particles and packed into a 0.5 m3
column. The total effectiveness factor for the system is close to unity. Km for the
immobilized enzyme is 1.32 kg/m3, Vmax is 45 kg/m3/hr. The lactose concentration
in the feed stream is 9.5 kg/m3. A substrate conversion of 98% is required. The
column is operated with plug flow for a total of 310 days/year.

a) At what flow rate should reactor be operated?

b) How many tonnes of glucose are produced per year?

Multistage Chemostat in Series

Q11. In a 2 stage Chemostat system the volumes of the I and II reactors are 500L
and 300L respectively. The I reactor is used for biomass production and the
second reactor is for secondary metabolite formation. The feed flow rate to the I
reactor is 100 L/hr and glucose concentration in the feed is 5 g/L

μm= 0.3 hr-1

YX/S= 0.4 g/g

KS= 0.1 g/l

a) Determine cell and glucose concentrations in the effluent of the I stage

b) Assume growth is negligible in the II stage and the specific growth rate of
product formation.

qp =0.02 g/g/hr and YP/S= 0.6 g/g

Determine the product and substrate concentrations in the effluent of the II

reactor.

Q12. Given:

μm= 0.7 hr-1

YX/S= 0.65 g/g

KS= 5 g/l

You want to cultivate this microorganism in either 1 fermenter or 2 fermenters in

series. The flow rate and substrate concentrations of inlet stream should be 500
L/hr and 85 g/L. The outlet substrate concentration is 5 g/L.
a) If you use one fermenter, what is the size of the reactor and outlet biomass
concentration?

b) If you use 2 reactors in series, what are their sizes?