Introducción a los

Bioreactores

Reactor Fed-Batch
Prof. Pablo Araujo Granda Ph.D.

paaraujo@uce.edu.ec
 Lecturas iniciales

• Documentos en aula virtual

• Cap. 13. Reactor Engineering - Bioprocess

Engineering Principles - Doran 1995
he knowledge base and its
roblems or disturbances. operating strategy has a signific
conversion, product concentration,
Modos de Operación
w information about pro- - Tipos
ledge base. To maximise nation and process reliability.
elligent supervisory con- Characteristics such as final subst
• Discontinuo
and representative concentrations and the time requir
data- Batch
and engineering knowl- determined for different reactor ope
• Alimentación
n rule formulation and intermitente
balances. –
For Fed-Batch
a general reaction syst
Expert systems can also change of component masses in the
• Alimentación Continua
tion - CSTR
using Eq. (6.5):
ting unmeasurable fer-
contradictory data and
dM
lism [30-33].
nce with applications in dt
neural networks. Neural
xtracting useful informa-
Alimentación intermitente (Fed-batch)
 differential
kAs
d isin the
e determined using
specific
for the
death
different product constant.
reactor ruleoperating
ofApplying
Eq. (D.22) in Appendix
these
schemes termsM,
using inis equal
mass __ = x ( t u
Section 13.5.1.2, mass ofliquid
concentration. By starting with cells
ly. Forin
volume the
the reactor,
V is not constant.
fed-batch reactor Equations
of
dt Figure
Eq.D. After
(6.5)
balances. grouping
gives: terms
I3 Reactor Engineering
Foradding
ax general this
reaction gives: d(sV) the fe
substrate and more nutri- system, F.we canliquid
relate rates ofoutrate
ds,change Reactor Fed-Batch - células
to xVwhere
of
high growth
of d(xV)
biomass
dV
component
is cell
rates are avoided.
generation
dx
concentration
masses in the
R G is processing,
F i - and
ture are
system
Vis
derived
With
to
The unsteady-state
equal to t,we the
dilute
rate
x V can
volume,
by carrying
of
where/u
solutions
assumereac-
mass-balance
p Let
is
unsteady-sta
such as
isthe
of
dtus equatio
constantnow ap
tho
cei
Fs
an
le,tion
in xusing
cultures Eq. (6.5):
where the oxygen k dferme
<<
Balance
specific 7- - Fxde
+
growth
Figure 13.21 l ~material
i +rate, = x Vand
Preparation, - kaxV.lag,
rate
reaction enofa
kd)xV.
can
and estado
flow
cell
harvest reactor
then
death
times in beNO
Rwas
taken
C
Figure
isestacionario
derived
13.22 outside
equal in
to
Flowsheet Chapter
batch
kdXV
for a of the differ
stirred 6:
reactor.
fed-batch
is too high d t for the mass-transfer
operation of a batch fermenter.
rateFeedof;tream
substra
w dM h through
e the equation.
r (13.36) Therefore, e Eq. (6
lternatively, high substrate con- d(pV) (13.35) where/~ ofsubstrate is thedxins
k ddt mentation is:
= pi-
theseopo
tory oris switch
the specific on undesirable death constant. dtApplying termsyield
plied
infrom substr
by volum__
Applying
chBecause
Eq.
culture isEq.
(6.5)Vin (13.34)
fed-batch
gives:
used extensively toculture
Eq. (13.36):in is a function of time, it cannot M._ tion,
dt
I I
(6.5) .) YPS iscoupl
directly the tr
o be Considerando
cancelledcatabolite
overcome =
xf
through las células Eq.
repression(13.35).
I
Instead,
I dV we must expand the maintenance co
r -F. (13.21). Subst
Let u
differential
itwhere
is also used dx
using
routinely the product
for peni- rule of
where Eq.dt p (D.22)
is densityin of F
Appendix
the= Caudal volumétrico
reactor contents,
applying = cte
Eqs Vi
(13
d(xV)
x FMis
+ Vmass =of Fcomponent
x i + ( / a - k d ) A
x V in. the vessel, t is time, A7ri is
D. After -
grouping
d
8
t Fx i +
terms l ~ x V
this - kaxV.
gives: V = Volumen liquido = no batch
cte
the mass d t flow rate of A entering the reactor, M ois the mass flow dsd(sV)
rate of A leaving,
Xo "11
tpt~. t lR G istb the thvmass
I --V"
,~ tplb.-Itlrate
i
ofgeneration of(13.37)
A byFo = reac-
0 rate o
Fi = F (13.35) dt D(s i - sF
~ -------- ~ tb-s thv
dV R C is the
tion,x and dx mass tT
rate -"
of A
tT similar
consumption "- mass
of A balance
by reaction.based on dt Eq. ofsub (6.5)
7- + = Time
kd)xV.
cells.gives:
In Eq. fed-batch
Dividing through by
In this section we consider application Vand rearranging of (6.5) tooperation
batch, M o = plied 0; M
Because Vin fed-batch culture flow is a function of time,
rate F multiplied
(13.36)it cannot
by the cell concent
where/~ direct
isand
the
be cancelled
dx D F through Eq. (13.35). Instead, we must expand Eqs
As in Section 13.5.1.2, mass of cells (13.2 (13.41)
the in th
- ntion
x ioccurs.
+ These x time -periods
k d - -are illustrated
~- . for fermenta- yield from
change of cellsub
an
differential
dt
Applying Eq. V using
(13.34)
tion processes
the product
to 13.21.
in Figure Eq. Therefore,
(13.36): rule
to of
xVwhereEq.
the total downtime
(D.22)
x is cellin Appendix
concentration
Densidad = cte = diluído and V
tion, YPS is the
D. After grouping terms this gives:
tdn associated with batch reactor operation is:
of biomass generation R G is equal to t
 applying Eqs (13.3
xF+ V = F x i + ( / a constant,
-kd)xV. D decreases as the reaction procee
I3 Reactor Engineering
dt
Reactor Fed-Batch - células (13.39) to Eq. (13.38):(13.37) ds
D(s i - s )
dt
nd Fo Balance
are input Figure (13.34)
de
and output material
mass
13.21 Preparation,flow dx
rates,
lag,
en estado
and
reaction
NO estacionario
and harvest times in Figure 13.22 Flowsheet for a stirred fed-batch ferm
Dividing throughoperation ofby Vand
a batch fermenter.rearranging gives:
nsities of input and output streams, respective- d--t - Let Dxius define
+ x the
(/~dilution
- k d -rate DDwith ). Feed ;tream
-batch reactor of Figure 13.22, Fo - 0 and dimensions T
can dxbe performed
lute solutions F
such as those
for
often used in bio- Eqs (13.41) and (
D
- nisxconstant
i + the and xfeed - kPid=- P;- density
~- . F
i is
an equal
assume
dt p to
V that DI ~ m~ change of cell and
M._ .)
I

ken outside of the =differential

xf and cancelled V
tration x i in the feed. I I

uation.Velocidad
Therefore, de dilución:
Eq. (6.6)Dfor
r
[tiempo Eq.
fed-batch -1] (13.40)
fer- can be simplified for most appl
e reactor, M, is equal
8 the feed material In fed-batch is sterile
systems so that xwith
V increases i = 0.time;
If, ther
in
is liquid volume, rate I constant, D decreases as the reaction proceeds. A
Xo "11 of cell
--V" death i
tb-s (13.39)
is negligible compared with
t, x V where/u is the tpt~. t l ~ -------- t b thv ,~ tplb.-Itl~ thv to Eq. (13.38):
tT k d
-" << ju,
tT Eq. "- (13.40) becomes:
h R C is equal to kdXV Time (13.34) dx
Simplificaciones: d--t - Dxi + x (/~ - k d - D ).
r on Eq. (6.5) can ebe performed
alance based dx for
1. Material esterilizado xi = 0
lying these
ch operation
2. Kd M ou =terms
<<<< 0; M i isinequal to the __feed= x ( t u - D ).
tiplied by the cell
tion concentration
occurs. These time periods x i in the aredt feed. for fermenta-
illustrated
tion processes in Figure 13.21. Therefore, the total Eq.downtime
(13.40) can be simplified for most applicatio
3.5.1.2, mass oftdncells in the reactor, M, is equal
associated with batch reactor operation is: the feed material is sterile so that x i = 0. If, in addit
cell concentration and Vis liquid volume, rate
 Let usfor now apply Eq. reactor
(6.5) to the limiting
derived insubstrate in using
our fed-mass
tedetermined
in rates of substrate
concentration.
the feed.
Chapter 11.batch
uptake
Eq. (13.40)
different
By and product
canstarting
be
formation
with
simplified
operating
forand liquid
most
schemes
volume
applications. Vmass
Usuallyis not constant. Equations
M,balances.
is equal I3 reactor.
Reactor In
Engineering this case,/t~/o RGare zero; the flow
substrate andFor amaterial
addinggeneral more reaction
so that xsystem,
nutri- inwe can therelate rates ofout unsteady-st
Reactor Fed-Batch - Sustrato
the
Let us apply feed
Eq. (6.5) to the isgrowth-limiting
sterile i =ture
0. If, are
substrate in addition,
a derived byratecarrying
olume, rate rate of substrate entering the reactor M i is equal to Fs i. Mass
eds,change
batch
(13.35) high ofofcomponent
growth cell AT/i
fermentation. death= 35Iis
rates are masses
0 =negligible
0avoided. in
because comparedthe
substrate
re/u is the ofsubstrate in the reactor, M, is equal to concentrationand
system
with
does
The to
growth
notreactor,
the thesoF rate
unsteady-state ithat
s
of reac-
Fo mass-balance
are
multi- input and output equatioma
flow into or out k d of
<<the
ju, reactor;
Eq. (13.40) the mass becomes:
of substrate in the reac-
ple,
ual tion
to in
kdXVusing
cultures
plied Eq.
by (6.5):
where
volume theV. oxygen
For fermentations a flow Pi and Po are
reactor
producing wasdensities
product derived
notof input and output str
infor Chapter 6:360
cannot e
isliquid
Balance de material en estado NO estacionario
tor, M, is equal to s Vwhere s is substrate concentration and Vis
toovolume.
high
Figure
for
13.21
operation
directly the
dx coupled
Substrate
Preparation,
mass-transfer
of aisbatch
not fermenter.
lag,
with energy
generated;
reaction and harvest
metabolism,
therefore
times in Figure 13.22
R G = 0. R C R e is given by Eq.
Flowsheet
ly. For the fed-batch reactor of Figure 13
a stirred fed-batch fermen

eand the into r s V __

isterms = x ( t u - D ). F i - F. With dilute solutions Feed ;tream
such as those o
dM (13.21).
equal
Alternatively, where
dthigh r s is
Substituting the
substrate volumetric
these
con- rate
terms of substrate
into Eq.
d(pV) (6.5) gives:
(13.41)
ppendix
uptake. As discussed in Section 11.10, the expression for processing,
rs we can assume p is constant and th
itory dtoron whether
depends switch on undesirable
extracellular product is formed by the dt
cul- = pi- opo
Let us now apply Eq. (6.5) to the limiting can
d(sV) thenin be
substrate our taken
fed- outside of the differenti
atch
ture culture
input and
andthe
output ismass
batch used
relationship
reactor. extensively
between
flow rates,
In product
this and
I in
/a synthesis
case,/t~/o and and energy
qPRGare I
s) the mass
through
zero; the equation.
flow M._ .)
Therefore,
(6.5) (13.38)
Eq. (6.6) f
input
Considerando
generation in
and output the xf
cell.
dtof el
If
streams,
= sustrato
Fs i -
product
respective- isI limitante
Yxs
formed+ but y
notproducto
+ m
directly
I
x V no
to overcome ratecatabolite
substrate repression
entering rvs
the reactor M i ismentation
equal to Fsis:
i. Mass
asociado
coupled
(13.35)
eactor with
of Figure al13.22,
energy metabolismo
metabolism,
ofsubstrate
r
Fothe
in
r s
- reactor,
0 and is energético
given by Eq. - PNAME
(11.76)
Let us define
M, is equal to
and:
the dilution rate Dwith
concentration s multi- (13.42)dimensions T - 1:
F = Caudal volumétrico = cte
;tions
it issuch
where alsoMis
usedmass
routinely
of for
component peni- A in where p is density
the vessel, of theA7r
t is time, reactor
i is contents, V
e,(13.36)
RcrsV(
it cannot
me the mass
as those
flow
-- where/~
p is constant
expand the
often
plied by

ide of the differential

rate of
Appendix A (13.21).
yield
leaving,
8
=and that
directly
YXS
from
used
volume
rate
is
Xo "11
tpt~. t
in For
V.
Pi
coupled
and
bio-fermentations
+the=ofP;specific
Substituting
l ~R YPS
substrate,
G
A
cancelled
-------- tis
b
)
producing
+entering
density
with
m Senergy

the
I
these
qp
thv
product
growth
is
mass
,~
not

--V"
terms
the
tplb.-Itl~
the
x V metabolism,
D

into
specific
rate
F
~ reactor,
rate, m ~ Yxs
V (6.5)
Eq. i
rate
ofgeneration
tb-s thv
dV
R eisisM
dtof
theois
given
gives:
product
the
- Ftrue
.by
of
mass
biomass
Eq. V = Volumen
flow
Fo = reac-
forma-
A by 0
liquido = no cte

(13.21) (13.39)
herefore, Eq. (6.6)
tion, YPS foris fed-batch
thetTtrue product fer- -" yieldtT from substrate
"- and m s isFithe =F
tion, and Rd(sV) C is the mass rate Time of consumption of A by reaction.
maintenance coefficient. /a InExpanding
qP
fed-batch the differential
s)systems V increases with and time; therefore, if F is
In this applying
section dt Eqswe (13.34) consider
Fs i -
and application
Yxs +
(13.39)
rvs
constant,
+ m
gives:
Aof x V Eq.
similar (6.5)
mass to
balance batch,
based on Eq. (6.5) can
D decreases as the reaction proceeds. Applying Eq.
cells. In fed-batch operation M o = 0; M i is
(13.39) to Eq. (13.38): (13.42)
(13.36) ds flow rate F multiplied by the cell concentrati
(13.37) where/~ isThesethe specific t~ rate, Yxs
growth qP is the s)true biomass
tion occurs.D(s i - s ) - (13.34)
time periods are
YXS + dx
illustrated As
+ in
for fermenta- m Section
x. 13.5.1.2, mass of cells in the rea
Densidad = cte = diluído
dt processes
yield
tion from substrate,
in Figure 13.21. qp isTherefore, d--tthe- rps
the specific rate
total
Dxi of product
downtime
to + x (/~
xVwhere - k forma-
d x- isDcell
). concentration and Vis liq
tdn associated
tion,canYPS with batch reactor operation is:
ased on Eq. (6.5) beisperformed
the true product for yield from substrate and m s is the (13.43)
Simplificaciones – PNAME – Fed-Batch

• D = f(t) .. Integrar .. Se complica L

• Operación del reactor

• 1ero discontínuo hasta [células] elevada y
[sustrato] casi desaparece
• 2do operación intermitente (se abre la llave) J
 stant so that dX/dt = O. From Eq
batch until a high cell density is achieved and t
Therefore, substituting/u --- D i
Simplificaciones
virtually exhausted.–When
PNAME – Fed-Batch
this(11.60):
Eq. condition is reache
•operation
Resultado:is started with medium flow rate F. As
~rnax$
concentration x is maintained D ~, high and approxi
• [células – x] se mantiene elevada yKs+ “casi”
S = cte
stant so that dX/dt = O. From Eq. (13.41), if dX/dt
!"
Therefore, substituting/u ---'Dyinto the Monod e
• ≈ 0 y eso causa que & ≈ junto con
!# Rearrangement of Eq. (13.44) gi
Eq. (11.60): concentration as a function of di
Monod
DK s
~rnax$
D ~, ~max -- D
Ks+ S
Let us assume that the culture do
 there is product formation,
At high cell densit tha
Let us assume thatenergy generation.
the culture does If
not maintenan
produce p
Simplificaciones – PNAME – Fed-Batch
ing the vessel is co
there is product neglected,
formation,Eq.that
(13.43) can
as~atit= is0. be sim
directly
Applyin
Mas consideraciones:
energy generation. If maintenance
1. No se genera producto
requirements
(13.46), we obtain
ds /zx
neglected,
2. Formación de producto Eq. (13.43)
asociado al procesocan
-
be
D(ssimplified
de generación to:
i -- S) de-- energía
3. Despreciar necesidades de mantenimiento
dt X-~ YxsSiYxs .
ds /zx
- D(s i -- S) --

dt Yxs
At high cell density in the reactor
For product synt
ing the vessel
• [células – x] se mantiene elevada isolism,
consumed
y “casi” = cte Eq. immed
(13.4
as~at = 0. Applying these for
expression relatio
pr
At high cell density in the reactor, virtually all su
• Sustrato que entra = consume ”instantáneamente”
(13.46), we obtain:
Assuming the feed
ing the vessel is consumed immediately; therefore
as~at = 0. Applying X-~
!" theseYxsSi
relationships
. P = with/u
YPs
• S << Si // ≈ 0 // & ≈ ' Si"
!#
(13.46), we obtain:
cal expressions for fed-batch fed-batch reactors, the total
i

ds /zx
13.41) and (13.43). Here, we will Consider D(s i the rate of increase
re Simplificaciones – PNAME – Fed-Batch
- -- S) --

the reactor is operated first indt dX/dt, where Xis Yxsequal to xV.
the reactor, F i and Fo are input and output mass flow rates, and
sity is achieved Piand and the
Po are substrate
!"
densities of input and
and (13.47)
output streams, dX/dt= 0:
withrespective-
• [células - x] “casi” cte = ≈0
his condition is reached, !#
ly. For the fed-batch
fed-batch reactor of36I Figure 13.22, Fo - 0 and Let us
edium• Volumen
flow rate FF.i - As
líquido F. With
aumenta
a result, At
dilute
con high
solutions
el
cell tiempocell
such density
dXas
= those in the
often
masad(xV) used reactor,
células in bio-
dVvirt
processing, we can assumeing thep isvessel
constantisand that Pi = P; density
consumed immediatel
ned high
aumenta and approximately con- - - -
can then be taken outside of thedtdifferentialdtand cancelled dt
= x ~ D

m Even
Eq.though
(13.41), if dX/dt
through ~. as~at
theO,equation.
~t = D. = 0. Applying
Therefore, Eq. (6.6) for these relationshi
fed-batch fer-
cell concentration remains virtually unchanged !&
-D •into
with Velocidad
dX/dt=the dethe
Monod
O, because aumento
mentation is: deincreases
expression
liquid volume biomasa
of withtotal
(13.46), time = '( )*()' + = -.
we inobtain:
!# In fed
fed-batch reactors, the total mass of cells also increases. consta
Consider dV of total biomass in the Eq.
!& the rate of increase reactor(13.49) can now be(13.39
in
• where
dX/dt, !#
≈ 0Xis equal to xV. Using
-F.
the results X-~
of Eqs YxsSi
(13.34) .
dt
and (13.47) with dX/dt= 0: X - X 0 at the start (13.34)
of liquid flo
dx
d--
dX d(xV) A dV
similar massdxbalance based on Eq. (6.5) can be performed for
--- = x ~ + V = For product
YxssiF. X - -synthesis
Xo + ( Yxsdirectly
si F ) coupl
tfb
dt dt cells.
dt In fed-batch operation M o = 0; M i is equal to the feed
dt(13.44)
olism,
flow rate F multiplied Eq.concentration
by the (13.49)
cell (13.47) xallows us to d
i in the feed.
Eq. (1
expression
As in Section 13.5.1.2, mass of cellsfor product
in the reactor, M,concentration
is equal
the fe
 operation is started withthere
mediumis product
flow rateformation,
F. As a result,th
energy generation. If maintenan
Resumiendo - Fed-Batch
concentration x is maintained
neglected,
stant so that dX/dt = O. From
high and approximately
Eq. (13.43)
Eq. (13.41), can
if dX/dt
c
be~tsim
~. O, =
• !", $ ↑ & Therefore,
' ↓ → *'+,-. /'01-.0'+,2".3,4".
substituting/u --- D into the Monod expression
Eq. (11.60): ds /zx
56 5: 5;
- D(s i -- S) --
• ≈0 ≈0 ≈0 dt Yxs
57 57 57 $
~rnax
D ~,
• Muerte celular = 0Ks+ S
(13.
At high cell density in the reacto
• Necesidades mantenimiento =ingdespreciable
the vessel is consumed imme
Rearrangement of Eq. (13.44)
as~at = gives an expression
0. Applying for substr
these relati
• Producto =concentration
ausente as a function of dilution rate:
(13.46), we obtain:
DK s
X-~ YxsSi .
~max -- D
(13.
 eactor stant
is operated first= there
so that dX/dt in dX/dt,
O. From Eq.where
is product Xisif equal
(13.41), dX/dt ~.toO,
formation, xV.
that Using
~t = itD.is dirth
Therefore, substituting/u
chieved and the substrate ---
and D into the At
Monod
(13.47) with high cell
expression
dX/dt= 0: densityof in
Resumiendo
Eq. (11.60):
ition is reached, fed-batch
energy - Fed-Batch
generation. If maintenance
ing the vessel is requirem
consum
neglected, Eq. (13.43)as~at can=be0.simplified
Applying th to:
Eq
low rate$F. ↑As&a 'result,
• !", ↓ →cell *'+,-.dX d(xV)
/'01-.0'+,2".3,4". dV Xd-
~rnax $ - - - (13.46),
= x ~we obtain: + V
h and approximately con- ds /zx
D ~, dtD(s i dt dt d
• 5 ≈if 7dX/dt ~.Ks+
13.41), S =$,D.' , / ≈ 2+0
O, ~t
dt
- --
5, 8,Yxs
S) --
7, 9
X-~ YxsSi ≠ 2+0
.
the Monod expression of (13.44)
• Aumenta V = Velocidad crecimiento disminuye
Rearrangement of Eq. (13.44)Eq. gives
(13.49) can nowforbe
an expression integrated
substrate
At high cell density in the
For reactor,
product wha
virtually
synthesis
concentration as a X
function -
of X 0 at
dilution the
• Producto = asociado a metabolismo energético start
rate: of liquid flow to giv
ing the vessel is consumed immediately;
olism, Eq. (13.47) a ing
ther
bio
DK s as~at = 0.X -Applying
- Xo + ( Yxsthese
si Frelationships
expression
) tfb for producwi
(13.44) of
~max -- D (13.46), we obtain: Assuming the feed doe
(13.45)
ple
an expression for substrate X-~ YxsSi . P = YPs Si"
where tfb is the fed-batch time afterprec
on rate:
Let us assume that the culture does not produce product, or, if
dP
ing. Eq. (13.50) indicates that, for Yxs
00 (a) Estimate the time required to reach stationary phase.
l~e (b) What will be the final cell density if the fermentation is
on stopped after only 70% of the substrate is consumed?
ns
ng
Ejercicio # 1 – Fed-Batch
13.4 Fed-batch scheduling
Nicotiana tabacum cells are cultured to high density for pro-
duction of polysaccharide gum. The reactor used is a stirred
tank, containing initially 100 litres medium. The maximum
er- specific growth rate of the culture is 0:18 d-1 and the yield of
he biomass from substrate is 0.5 g g-l. The concentration of
er- growth-limiting substrate in the medium is 3% (w/v). The
to reactor is inoculated with 1.5 g l- 1 cells and operated in batch
he until the substrate is virtually exhausted; medium flow is then
started at a rate of 4 1 d-1. Fed-batch operation occurs under
quasi-steady-state conditions.
(a) Estimate the batch culture time and final biomass concen-
tration.
(b) Fed-batch operation is carried out for 40 d. What is the
of final mass of cells in the reactor?
ric (c) The fermenter is available 275 d per year with a downtime
to between runs of 24 h. How much plant cell biomass is
produced annually?
 Ejercicio # 2 – Fed-Batch
13.5 F e d - b a t c h p r o d u c t i o n
culture
of cheese starter fermenter. The biomass yield from substrat
0.7 mg l- 1, and the maximum specific gro
The medium contains 4% (w/v) methanol
LactobaciUus casei is propagated under essentially anaerobic
sion of 98% is desirable. The reactor may
conditions to provide a starter culture for manufacture of Swiss
batch or continuous mode. If operated in b
cheese. The culture produces lactic acid as a by-product of ener-
0.01% (w/v) is used and the downtime betw
gy metabolism. The system has the following characteristics:
If operated continuously, a downtime of
Yxs = 0"23 kgkg -1 year. Neglecting maintenance requirements
Ks = 0.15 kgm -3 al biomass production from batch and cont
jUmax = 0.35 h -~
ms = 0.135 kgkg -l h -l
13.8 Reactor design for immobil
A stirred fermenter is operated in fed-batch mode at quasi-
6-Aminopenicillanic acid used to produce se
steady state with a feed flow rate of 4 m 3 h-1 and feed sub-
lins is prepared by enzymatic hydrolysis of f
strate concentration of 80 kg m -3. After 6 h, the liquid
penicillin G. Penicillin acylase immobilised
volume is 40 m 3.
considered for the process; the immobilised
(a) What was the initial culture volume? small enough that mass-transfer effects can b
(b) What is the concentration of substrate at quasi-steady ing concentration of penicillin G is 10% (
state? high cost of the substrate, 99% conversio
(c) What is the concentration of cells at quasi-steady state? these conditions, enzymatic conversion of
(d) What mass of cells is produced after 6 h fed-batch opera- considered a first-order reaction. It has not b
tion? a batch, CSTR or plug-flow reactor would b
downtime between batch reactions is expect