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Enzyme reactor design

Mahesh Bule
Levenspiel’s four fundamental
questions
• In approaching the design of a reactor system the engineer has to
answer a number of important priliminary questions before
embarking on his detailed calculations. These questions are as
follows:
1. Do I have the right reactor type in mind: should it be plug flow,
mixed flow, recycle, multistage or what?
2. What temperature progression should I aim for: constant, rising,
falling etc. and should that require heat exchange, may be
multistage?
3. For a catalytic reaction what size of particle should be used? This
tells what type of reactor should be used: packed bed, fluidized
etc?
4. Does the catalyst deactivate and if so, does it deactivate rapidly or
slowly?
Platform to simulate and optimize the
enzyme reactor operation

BR – Batch reactor,
BRP – Batch reactor with intermittent addition of enzyme
SBR – Semi-batch reactor,
MACR – mechanically agitated continuous reactor,
FXBR – Fixed bed batch reactor
Reactor Types
• Ideal
– PFR
– CSTR
• Real
– Unique design geometries and therefore RTD
– Multiphase
– Various regimes of momentum, mass and heat
transfer
Reactor Cost
• Reactor is
– PFR
• Pressure vessel
– CSTR
• Storage tank with mixer
• Pressure vessel
– Hydrostatic head gives the pressure to design for
Reactor Cost
• PFR
– Reactor Volume (various L and D) from reactor kinetics
– hoop-stress formula for wall thickness:
PR
t  tc
– SE  0.6 P

• t= vessel wall thickness, in.


• P= design pressure difference between inside and outside of
vessel, psig
• R= inside radius of steel vessel, in.
• S= maximum allowable stress for the steel.
• E= joint efficiency (≈0.9)
• tc=corrosion allowance = 0.125 in.
Reactor Cost
• Pressure Vessel
– Material of Construction gives ρmetal
– Mass of vessel = ρmetal (VC+2VHead)
• Vc = πDL
• VHead – from tables that are based upon D

– Heat capacity Cp= FMCv(W)


Reactors in Process Simulators

• Stoichiometric Model
– Specify reactant conversion and extents of
reaction for one or more reactions
• Two Models for multiple phases in chemical
equilibrium
• Kinetic model for a CSTR
Used in early stages of design
• Kinetic model for a PFR
• Custom-made models (UDF)
Mass Balance on Reactive System
• In - out + gen - cons = accumulation
FA0 FA
Rate of flow in Rate of flow out
System

GA
Rate of
generation/
consumption

• A mass balance for the system is


dN A
FA0  FA  G A 
dt
• NA is the mass of “A” inside the system.
• The reaction term can be written in more familiar
terms,
GA = rA V
• V is volume of the system.
• Note that the units for this relation are consistent:
mass mass
  volume
time volume  time
• If GA (and hence rA) varies with position in the
system volume, we can take this into account by
evaluating this term at several locations. Then
DGA1 = rA1 DV1,
• Summing the reactions over the entire volume
yields:

k k
G A   DG Ai   rAi DVi
i 1 i 1

• As k   (that is, as we decrease the size of these cubes


and increase their number)
• DV  0 which gives

V
GA   rA dV
Generalized Design Equation for
Reactors

• In - out + gen - cons = accumulation

V
dN A
FA0  FA   rA dV 
dt
Types of Reactors
• Batch
– No flow of material in or out of reactor
– Changes with time
• Fed- Batch
– Either an inflow or an outflow of material but not both
– Changes with time
• Continuous
– Flow in and out of reactor
– Continuous Stirred Tank Reactor (CSTR)
– Plug Flow Reactor (PFR)
– Steady State Operation
Batch Reactor
• Generalized Design Equation for
Reactors V
dN A
FA 0  FA   rA dV 
dt
• No flow into or out of the reactor,
then, FA = FA0 = 0
dN A V
  rA dV
dt
• Good mixing, constant volume

dN A d N A V  dC A
 rAV or   rA
dt dt dt
Enzyme Batch Reactor
(constant volume, well mixed)

dS vmax S
r 
dt K M  S
• integrate from t = 0 to t = t, we obtain
Kmln (S0/S) + (S0 -S) = vmax t

• Batch reactors are often used in the early stage of


development due to their ease of operation and
analysis
Batch Enzyme Reactor Determination of
M-M kinetic
Linear form becomes parameters
(S0 – S) t
= - KM + Vmax
ln(S0/S) ln(S0/S)

(S0 – S)
ln(S0/S)

Vmax

t
- KM
ln(S0/S)
Fed Batch Reactor
• Reactor Design Equation
V dN A
FA0  FA   rA dV 
dt
• No outflow FA = 0
• Good Mixing rA dV term
out of the integral

dN A d C A V 
FA0  rA V  
dt dt
Fed Batch Continued
• Convert the mass (NA) to concentration. Applying
integration by parts yields
dC A dV
FA0  rAV  V  CA
dt dt
• Since dV
 FA0
dt
• Then
dC A
FA0  rAV  V  C A FA0
dt
• Rearranging

dC A FA0 C A FA0
  rA 
dt V V
Fed Batch Continued
• Or
dC A FA0
 1  C A   rA
dt V

• Used when there is substrate inhibition and


for bioreactors with cells.
Assumptions for a fed batch reactor
include
1. Only a feed in
64%
2. Either a feed in or a
removal stream
3. Steady state 29%

4. 2 and 3
7%
5. All of the above 0% 0%

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Continuous Stirred Tank Reactor

• Assume rate of flow in = rate of flow out


• FA = v CA and FA0 = v CA0
• v = volumetric flow rate (volume/time)
CSTR - continued
• General Reactor Design Equation
V dN A
FA0  FA   rA dV 
dt
dN A
0
• Assume Steady State dt

V
• Well Mixed  rA dV  VrA

FA0  FA
• So FA0  FA  VrA  0 or V
 rA
CSTR for Enzymes
(Enzyme remains inside)
• Input - output + generation - consump = accumulation
dS
FS0  FS  rV  v
dt
• F - flow rate l/hr
• S - substrate conc.
• V- reactor volume
• r - reaction rate
• at Steady State dS/dt = 0
CSTR - enzymes
rV = F(S0 - S)
and
vmax S
r
KM  S
Introducing space-time θ ( = V/F) and r in above equation we get

VmaxS
S0 =S + θ
KM + S
Continuous Stirred Tank Enzyme Reactor at steady-state
Linear form becomes

CS = - KM + Vmax (42)
(S0 – S)

S
Determination of
M-M kinetic
parameters
Vmax


- KM
(S0-S)
Plug Flow Reactor (PFR)

• Tubular Reactor
• Pipe through which fluid flows and reacts.
• Poor mixing
• Difficult to control temperature variations.
• An advantage is the simplicity of construction.
PFR Design Equation for Product
formation
• Design Equation V dN A
FA0  FA   rA dV 
dt
• Examine a small volume element (DV) with length
Dy and the same radius as the entire pipe.
Flow of
Flow of
A out of
A into
Element
Element
• If the element is small, then spatial variations in rA
are negligible, and Assumption of “good
V mixing” applies only to

 rA dV  rA DV the small volume


element
• If volume element is very small, then assume steady
state with no changes in the concentration of A.
dN A
0
dt
• Simplify design equation to:
FA  y   FA  y  Dy   rADV  0
• rA is a function of position y, down the length of the
pipe and reactant concentration
• The volume of an element is the product of the
length and cross-sectional area,
DV = A Dy
• Design Equation becomes:
 FA  y  Dy   FA  y 
   ArA
 Dy 
• take the limit where the size of a volume
element becomes infinitesimally small
dFA
lim  ArA
Dy  0 dy

• or because Dy A = V,
dFA
 rA
dV
• This is the Design Equation for a PFR
• Bioapplications - Sometimes hollow fiber
reactor analysis is simplified to a PFR
Plug-flow Enzyme Reactor at steady-state

F F F F
S0 S S+dS Sf

dV
Mass balance for the substrate over dV:
FS = F(S + dS) + (-rS) dV

The above can be simplified to - FdS / dV = -rS

F for the steady flow rate through the reactor


S for concentration of the substrate
dV for small volume of the reacting mixture
(-rS) for substrate utilization rate in dV
Plug-flow Enzyme Reactor at steady-state

F F F F
S0 S S+dS Sf

dV
Introducing space-time θ ( = V/F), we get

- dS / dθ = -rS

F for the steady flow rate through the reactor


S for concentration of the substrate
dV for small volume of the reacting mixture
(-rS) for substrate utilization rate in dV
Plug-flow Enzyme Reactor at steady-state
Substituting (-rS) for the simple enzyme reaction in, we get

dS VmaxS
- =
dθ KM + S
Rearranging above equation we get
S θ

S0
∫( -
KM + S
S ) ∫ dS =

0
Vmax dθ

Integrating above gives

()
KM ln
S0
S
+ (CS0 – CS) = Vmax θ
Plug flow enzyme reactor Determination of
M-M kinetic
Linear form becomes parameters
(S0 – S) θ
= - KM + Vmax
ln(S0/S) ln(S0/S)

(S0 – S)
ln(S0/S)

Vmax

θ
- KM
ln(S0/S)
Immobilized enzyme reactor (example)

Recycle packed column reactor


Advantages of immobilized enzymes:
- Easy separation from reaction mixture, providing the ability to control reaction
times and minimize the enzymes lost in the product

- Re-use of enzymes for many reaction cycles, lowering the total production
cost of enzyme mediated reactions

- Ability of enzymes to provide pure products

- Possible provision of a better environment for enzyme activity


Disadvantages of immobilized enzymes:
- Problem in diffusional mass transfer
- Enzyme leakage into solution
- Reduced enzyme activity and stability
- Lack of controls on micro environmental conditions
Methods of immobilization

1) Entrapment Immobilization
2) Surface Immobilization
3) Cross-linking
1) Entrapment Immobilization
It is the physical enclosure of enzymes in a small space.
- Matrix entrapment (matrices used are polysaccharides, proteins,
polymeric materials, activated carbon, porous ceramic and so on)
- Membrane entrapment (microcapsulation or trapped between thin, semi-
permeable membranes)
1) Entrapment Immobilization

Advantage is enzyme is retained.

Disadvantages are
- substrate need to diffuse in to access enzyme and
product need to diffuse out
- reduced enzyme activity and enzyme stability owing to the lack of control of
micro environmental conditions
2) Surface Immobilization
- Physical adsorption (Carriers are silica, carbon nanotube,
cellulose, and so on; easily desorbed; simple and cheap;
enzyme activity unaffected )
- Ionic binding (Carriers are polysaccharides and synthetic
polymers having ion-exchange centers)
- Covalent binding (Carriers are polymers containing amino,
carboxyl, hydroxyl, or phenolic groups; loss of enzyme activity;
strong binding of enzymes)
Methods of immobilization
3) Cross linking

is to cross link enzyme molecules with each other using agents such
as glutaraldehyde.
Comparison between the methods
Characteristics Adsorption Covalent Entrapment Membrane
coupling confinement
Preparation Simple Difficult Difficult Simple
Cost Low High Moderate High
Binding force Variable Strong Weak Strong
Enzyme leakage Yes No Yes No
Applicability Wide Selective Wide Very wide
Running problems High Low High High
Matrix effects Yes Yes Yes No

Large diffusional No No Yes Yes


barriers

Microbial protection No No Yes Yes


Immobilized enzyme reactor (example)
Recycle packed column reactor

- Allow the reactor to operate


at high fluid velocities
Immobilized enzyme reactor (example)
Fluidized bed reactor
- A high viscosity substrate solution

- A gaseous substrate or product in a


continuous reaction system

- Care must be taken to avoid the


destruction and decomposition of
immobilized enzymes
Immobilized enzyme reactor (example)

- An immobilized enzyme tends to Continuous stirred


decompose upon physical stirring.
tank reactor
- The batch system is generally suitable for
the production of rather small amounts of
chemicals.
Effect of mass-transfer resistance in immobilized
enzyme systems:

Mass transfer resistance is present


- due to the large particle size of the immobilized enzymes
- due to the inclusion of enzymes in polymeric matrix
Effect of mass-transfer resistance in immobilized
enzyme systems:

Mass transfer resistance are divided into the following:


- External mass transfer resistance (during transfer of substrate
from the bulk liquid to the relatively unmixed liquid film
surrounding the immobilized enzyme and during diffusion
through the relatively unmixed liquid film)
- Intra-particle mass transfer resistance (during diffusion from
the surface of the particle to the active site of the enzyme in an
inert support)
External mass-transfer resistance:
Assumption:
CSsSs
- Enzymes are evenly distributed on the
CSbSb
surface of a nonporous support material.
- All enzyme molecules are equally
active.
- Substrate diffuses through a thin liquid
film surrounding the support surface to
reach the reactive surface.
- The process of immobilization has not
altered the enzyme structure and the M-
M kinetic parameters (rmax, KM) are
unaltered. Enzyme

Liquid Liquid
Film Thickness, L
film thickness, L
External mass-transfer resistance:
Diffusional mass transfer across the liquid film:
CSsSs
CSbSb
JS = kL (CSb – CSs)

kL liquid mass transfer


coefficient (cm/s)

CSb substrate concentration in the bulk


solution (mol/cm3)

CSs substrate concentration at the


immobilized enzyme surface (mol/cm3)
Enzyme

Liquid Liquid
Film Thickness, L
film thickness, L
External mass-transfer resistance:
At steady state, the reaction rate is equal to the
mass-transfer rate: CSsSs
CSbSb

Vmax CSs
JS = kL (CSb – CSs) =
KM + CSs

Vmax maximum reaction rate per unit of


external surface area (e.g. mol/cm2.s)

KM is the M-M kinetic constant (e.g.


mol/cm3)
Enzyme

Liquid Liquid
Film Thickness, L
film thickness, L
External mass-transfer resistance:
Vmax CSs
JS = kL (CSb – CSs) =
KM + CSs

Non dimensionalizing the above equation, we get

1 - C’Ss β C’Ss
=
NDa 1 + β C’Ss

where
C’Ss = CSs / CSb

NDa = Vmax / (kL CSb ) is the Damköhler number

β = CSb / KM is the dimensionless substrate concentration


Damköhler number (NDa)

Maximum rate of reaction Vmax


NDa = =
Maximum rate of diffusion kL CSb

If NDa >> 1, rate of diffusion is slow and therefore the limiting


mechanism

rp = JS = kL (CSb – CSs)

If NDa << 1, rate of reaction is slow and therefore the limiting


mechanism
Vmax CSs
rp =
KM + CSs

If NDa = 1, rates of diffusion and reaction are comparable.


Effectiveness factor (η)

actual reaction rate


η=
rate if not slowed by diffusion

rmax CSs β C’Ss

KM + CSs 1 + β C’Ss
η= =
rmax CSb β

KM + CSb 1+β

Effectiveness factor is a function of β and C’Ss


Internal mass transfer resistance:
Assumption:
- Enzyme are uniformly distributed in
spherical support particle.
- Substrate diffuses through the
tortuous pathway among pores to
reach the enzyme
- Substrate reacts with enzyme on the CSs
pore surface
-Diffusion and reaction are
simultaneous CSr2

- Reaction kinetics are M-M kinetics


Diffusion effects in enzymes immobilized in a
porous matrix:
Under internal diffusion limitations, the rate per unit volume is
expressed in terms of the effectiveness factor as follows:

Vmax’ CSs
rS = η
KM + CSs

Vmax’ maximum reaction rate per volume of the support

KM M-M constant

CSs substrate concentration on the surface of the support

η effectiveness factor
Diffusion effects in enzymes immobilized in a
porous matrix:
Definition of the effectiveness factor η

reaction rate with intra-particle diffusion limitation


η=
reaction rate without diffusion limitation

For η < 1, the conversion is diffusion limited

For η = 1, the conversion is limited by the reaction rate

Effectiveness factor is a function of β and C’Ss


Diffusion effects in enzymes immobilized in a
porous matrix:

β
η

Theoretical relationship between the effectiveness factor (η) and first-


order Thiele’s modulus (φ) for a spherical porous immobilized particle for
various values of β, where β is the substrate concentration at the surface
divided by M-M constant.
Diffusion effects in enzymes immobilized in a
porous matrix:

Relationship of
effectiveness factor (η)
with the size of
immobilized enzyme
particle and enzyme
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