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Design Equation based on

Biochemical Reactions

Introduction

The design equations of bioreactors are

1.

To find the reaction time

2.

To find the reaction volume

Need mass balance equations of

1.

Substrate conversion

2.

Growth/biomass produced

3.

Products formation

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Enzyme Reactors

Batch reactor
Substrate being converted to product by enzyme.
Material balance for S

Rate of formation Rate of enzyme

of substrate
Reaction

dS
V
VRo
dt
Ro the activity of enzyme expressed in unit volume
in unit time

at So, t=0 and S(t)

S

S0

dS Ro dt

The rate of enzyme catalysed reaction is dependent

on substrate concentration, T ,pH and degree of
decay of that enzyme.
For given value of pH and temp, v ( sp rate of
reaction) by MM eq
Ro

Vmax,t S
Km S

Vmax,t Vmax,0 exp kde t

Vmax,t at any time by

kde the first order of kinetic constant
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Combining thee situation gives

Ro

Vmax,0S
exp kde t
Km S

Substitute into the above material balance to give

dS

Separating the variables

So

Vmax.0S
exp kde t dt
KM S

t
Km S
dS Vmax,0 exp kde t dt
0
S

And after integrating

1 exp kde t
S
K m ln o So S Vmax,0
kde
S
5

In the case of the decay of enzyme is negligible, then

which results in the
kde 0 and 1 exp kdet t
kde
expression:
S
K m ln 0 S0 S Vmax t
S

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Cell Batch Reactor or plug-flow fermenter

For a batch reactor

Growth of the cells

dx
(V ) rxV
dt

Consumption of substrate
ds
(V ) rsV
dt

Product formation
dp
(V ) rpV
dt

7
7

The analysis of plug-flow fermenter (PFF) is

analogous to the ideal batch fermenter
Turbular flow fermenter

The properties of the flowing will vary in both

longitudinal and radial direction.
The ideal turbular flow without radial variation is called
PFF
Eg packed-bed fermenter and multistage fermenter

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A plug flow reactor

Assumming that Monods law applies from growth
kinetics,

Mass balance on cells yields

XS
dX
X max
dt
S Ks

Mass balance on substrate gives

dS
1 max
dt

Xs
Yx / s S K s

With initial conditions

X =Xo and S=So

when t = 0
9

A plug flow reactor

Rewrite mass balance on cells

X Yxs S X o Yxs So

Insert (10) into (8)

ds max [ X o Yxs So S ]S

dt Yxs
S Ks

10
11

On the integrations
X Yxs S So
S
K S Yxs ln
X o Yxs So K S ln o

Xo

So
max t X o Yxs So
12
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Cont
Where
t AL / Fo

L and A are the length and cross sectional area of plugflow reactor, respectively

Fo volumetric in flowrate

11

A CSTR (continuous stirred-tank reactor)

The microbial population can be maintained in a state

of exponential growth over a long period of time by
using a system of continuous culture.
Continuous culture can be operated as chemostat or
as turbidostat.

Reading Assignment:
Find
i. What are chemostat culture and turbidostat culture in
term of operability.
ii. Discuss the significance of each each culture
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A Continuous Stirred Tank Reactor (CSTR)

The cell material balance (with no endogenous or death

rate constant)
FX o FX Vrx V

dX
dt

13

rx is the rate of cells growth in the fermenter and

dX/dt change of cells concentration with time = 0 for staedy state
operation of CSTR
Eq becomes

1
X Xo
rx

14

m residence time

13

A CSTR

If the input stream is sterile (Xo=0), the cell in a

CSTR are exponential growing (rx=X) and no
endogenous constant, eq (14) becomes
m

1
D

15

D is known as dilution rate and equal to the reciprocal of

the residence time

Hence, for steady state CSTR with sterile feed,

D

max S
S Ks

16

In other words, specific growth rate can be control

by changing the medium flow rate

If D is set greater
than max, culture
cannot reproduce
quickly then it is
washed out
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cont

From eq 16, S can be calculated with known

residence time and the Monod kinetics parameters as
S

Ks

17

m max 1

valid only m max 1

If m max 1, the growth rate is less then the rate of
cells leaving the outlet stream, consequently all the
cells in the fermenter will be washed out, therefore
eq 17 is not valid

15

cont

If the growth yield is constant (no endogenous

metabolism)
X Yxs So S

18

substitute eq 17 into 18 yields the correlation for X as

Ks
X Yxs So
m max 1

19

Ks
P Po Yps So
m max 1

20

16

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cont

Usually Yxs varies with the limiting substrate and

growth rate, hence it affects the inclusion of
endogenous metabolism, eq 16 becomes

21
22

D kd net
or D kd

From eq 19, when S

D( So S )

X
Yxs

Ks

m max 1

the eq becomes

23

substitute D kd in eq 23, then

17

cont

YX/S is not constant with growth curve since S

must be first incorporated into cell mass and
recognizes intracellular S as subcomponent of
biomass
Pirt (1975) derived the term that relates
maintenance effect on cell mass as YX/S(m)
Y X / S (G )

X
S G S m

growth

(21)
maintenance

The true growth is

Y X / S (G )

X
S G

(22)
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Rate of S consumption in batch culture is

dS dS
dS

dt dt m dt G

where

(23)

dX
X
dt
dS
X

dt Y xs ( AP )

and

X
dS

Y xs
dt G

(22)

kd
dS
X mX

dt
Y

m
xs

(23)

maintenance coefficient
as listed in the table

19

Maintenance Energy requirement for various microorganisms.

Micro-organisms

m, maintenance
coefficient (g
substrate/ g wt cell
mass/h)

Penicillium chrysogenum

0.022

Aspergillus nidulans

0.029

Aerobacter aerogenes (aerobik) 0.094

Aerobacter aerogenes
(anaerobik)

0.473

20

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Hence,

dS
dt
dS
dt

mX
.

Y xs AP

(24)

Y xs

(25)

Y xs
1

(26)

Y xs

subscript
m

= maintenance

= growth

where m is the maintenance coefficient based on S.

Yxs(AP) is apparent yield. When Yxs is written, it should
be interpreted as Yxs(AP). Yxs (with no endogenous) is
constant, Yxs(AP) varies with growth condition if kd>0
21

m is usually very low and could be determined by

plotting 1/Ym vs 1/ to give straight line with slope
m and intercept, 1/YG.

slope
m=kd/Yxs

1/Ym
1/YG

1/D
(jam)
Graphical approach to estimating Yxs and m for chemostat

22

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max S
In presence of endogenous metabolism, i.e net K S kd
s
hence,

K s D kd
m D kd

X Yxs So S .

27

D
D kd

and

28

The balance product formation

29

DP q p X

q p is specific rate of product formation

23

For substrate balance

D So S

30

The biomass balance is unchanged from the case

with endogenous metabolism and yields
S

1
1
D kd X q p X
Yxs
Yps

K s D kd
m D kd

Eq (30) can be solved for X

D
X Yxs So S
D k q Yxs
d
p

Yps

31
24

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Productivity of Product and Cells biomass

The productivity of product and cells biomass (Prx)

can be found from DP and DX, respectively. The
dilution rate that maximizes productivity is found by
differentiating DP or DX with respect to D and setting
the derivative equal to zero.(will explain this method
later in graphical method)
The optimal value of D(Dopt) will depend on whether
endogenous metabilism and/or product formation are
considered. When kd=0 and qp=0, Dopt for biomass
production becomes

Ks
Dopt m 1

K s So

32
25

So >>>Ks, Dopt will approach D=max or the washout

point. Stable D max is very difficult, unless the flow
rate and liquid volume can be maintain exactly
constant

Consequently, a value of D slightly less than Dopt may

be a good compromise between stability and biomass
productivity. It should also be apparent that Dopt for
biomass formation will not necessarily be optimal for
product formation.

26

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Determination of maintenance coefficient is normally performed

using chemostat technique due to difficulty to differentiate in
batch

Theoretical biomass Yield is through

No of electron in substrate concept

Constant Yield on ATP concept

Method of electron no in substrate

it is found 3.14 0.11 g ash free dry wt is formed per electron.

No of mole of O2 required to oxidize substrate C to form CO2
and H2O times 4 (i.e 4 e is transfered to 1 mole O2)
i.e
C6H12O6 + 6O2 6O2 + 6H2O

27

Hence, glucose has 24 e per mol and if the product is not

formed during fermentation, the maximum biomass
formed by 0.42 g/g substrate can be achieved.

constant Yield on ATP

10.5 g dry wt cell can be formed per 1 mole of ATP

generated during catabolism cell reaction. The value is
different for every organism between 6 and 29 g mole
ATP-1.

28

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