Professional Documents
Culture Documents
CG4017
Bioprocess Engineering 2
Denise Croker, BM-028
Denise.Croker@ul.ie
Course Structure/Assessment
Course structure
Lectures/tutorials:
3 lectures/tutorials week
Labs:
Assessment
Final exam
Labs
70%
30%
04/09/2014
Lab Sessions
4 Experimental Labs
EXP A: Determination of KLa
EXP B: Operation of a Lyophiliser
EXP C: Operation of a Bioreactor
EXP D: TBC
Complete 2,
Submit 2
3 Computational Labs
Complete as many as you can
EXP E: Fermentation simulation-Superpro Submit 2 ( 1 of each)
EXP F: Fermentation simulation-Polymath
EXP G: Activated Sludge Process simulation-Polymath
EXP H: Diffusion in a microbial film simulation-Polymath
Lab Safety
Lab Safety Guidelines available on the server.
04/09/2014
Syllabus
Revisit Bioprocess Engineering 1 (CG4003)
Energy balances revisited. Heat transfer & heat exchanger design for
biochemical processing.
Learning Outcomes
On completing this module you should:
1.
2.
3.
4.
5.
6.
7.
04/09/2014
Textbooks
CG4017 Outline course notes available from the CES server (DCroker) & SULIS
Recommended: P.M. Doran, 2012, Bioprocess Engineering Principles, Academic
Press, ISBN: 9780122208515. Available in print and electronic formats 65.
M. L. Shuler, and F. Kargi, 2001, Bioprocess Engineering: Basic Concepts, 2nd ed.,
Prentice Hall, ISBN: 0-13-081908-5. (ca. 97 hardback).
1. REVIEW BIOPROCESS
ENGINEERING
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Bioprocess engineering essentially began with the requirement for industrial scale
production of antibiotics.
Penicillin discovered in 1928 by Alexander Fleming (UK). Discovery lay dormant for over
a decade.
Renewed interest during World War 2 to treat infection from battlefield wounds: clinical
trials very impressive.
Large scale production initially very difficult due to:
- low product concentration (only 0.001 g/dm3 !) in final fermentation broth
- requirement for large volumes of sterile air for the aerobic fermentation
- necessity for aseptic fermenter operation
- fragile nature of penicillin: recovery & purification challenges
Problems solved by US companies such as Merck, Pfizer, and Squibb: strain & fermenter
improvement gave over 50 g/dm3 product conc. Fermenter volumes of 40,000 dm3
used to meet capacity for treatment of 100,000 per year by 1945.
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Upstream Processing
Full Scale Bioreactor, 30,000L
< 25 Hours
Frozen Mother
Cell Strain
Shake Flask,
5 -6 hours
Seed Reactor
6-8 hours
Process Checks
pH, dissolved Oxygen, temperature, agitation rate automatically
Cell density Off line Test aseptic sampling loop required
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Downstream Processing
Isolate product from the fermenter
Product = 50 70 Kg of Sludge
All expressed proteins, cell debris
Biochemical Processes
Mechanically fragile
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Numeracy
Understanding of biochemical systems
Knowledge and practice of process engineering
Conceptual/design/original thinking capabilities
Current status of this field can be classified by type of biochemical activity:
1. Microbial (fairly mature technology, with some new developments)
2. Animal cell culture (newer)
3. Plant cell culture (newer)
4. Genetically modified organisms (newer)
5. Medical applications: tissue engineering, gene therapy etc. (very new)
6. Mixed cultures: food products, waste treatment, etc. (old, but poorly understood,
yet important, technologies)
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https://www.youtube.com/watch?v=LiYT5b3CsLk&index=4&list=PLUSfjij8XMn7mLVIGnBoaeUf1zMBrK9HW
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2. BIOCHEMICAL KINETICS
2. Biochemical Kinetics
A quantitative knowledge of biochemical kinetics is essential in order to design,
size, and predict the efficiency of bioreactors:
Bioreactor size
Speed of slowest kinetic step
2.1 Quantifying biochemical kinetics
Biochemical systems are complex in two major respects:
1. Structural complexity:
http://teachernotes.paramus.k12.nj.us/nolan/cp%20bio.htm
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2. Segregational complexity:
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Cells
4
3
Substrate molecule
Cells
4
3
2
Substrate molecule
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Cells
4
3
Substrate molecule
Cells
4
3
2
Substrate molecule
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Cells
4
3
2
Substrate molecule
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Reaction
type #
Zone(s) involved
Reaction
type #
Zone(s) involved
(1)
15
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Reaction
type #
Zone(s) involved
(1)
(2)
C+D
Reaction
type #
Zone(s) involved
(1)
(2)
C+D
(3)
B+C+D
16
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Reaction
type #
Zone(s) involved
(1)
(2)
C+D
(3)
B+C+D
(4)
B+D
Reaction
type #
Zone(s) involved
(1)
(2)
C+D
(3)
B+C+D
(4)
B+D
(5)
A+ D
A+ C+ D
A+ B+ C+D
A+ B+ D
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s
ds
max
dt
Km s
(1)
s = concentration of substrate
max = maximum rate at high s
(2)
Time dependence of
substrate concentration:
max s
Ks s
(3)
ds max qP
mS xo e max t
dt YXS
YPS
(4)
dp
qP xo e max t
dt
(5)
dx max s
x
dt K s s
(6)
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The relationship between microbial growth yield and substrate consumption (yield
coefficient relationship) is:
x xo
(7)
YXS
so s
max YXS so xo x
dx
x
dt ( K sYXS YXS so xo x)
(8)
Integration of (8) yields a sigmoidal cell growth equation, graphically depicted in the
previous slide. Practical use of an equation such as (8) requires a predetermined
knowledge of the maximum cell amount produced, xmax, in a given reaction
environment. xmax is identical to the ecological concept of carrying capacity.
k 1
xmax
(10)
dx
x
k x1
dt
x
max
(11)
Growth models for filamentous organisms: Here the organisms grow as microbial
pellets in submerged media, or as mold colonies on moist substrate surfaces. In
these cases the (normally linear) growth rate is expressed in terms of pellet or
colony, radius (R) or mass (M). Thus in the absence of mass transfer limitations:
2
dM
k p 4 R 2 M 3
dt
(12)
= pellet/colony density
= kp(36)1/3
N s
aL s
max
s
s
(13)
R s
a s
max
s
s
(14)
= effectiveness factor
a = area of active microorganism/unit film or floc volume
L = film thickness
= floc density
, = biological rate coefficients
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k1 [e]L[ S ]
1 k3 [ S ]
k1 [e][ S ]
= effectiveness factor
k1
max
Km
k 2 max
K
D
m e
k3
1
Km
(16)
= particle density
L = sheet thickness
0.5
(1 k3 [ S ])
(15)
Ns
ds
k s a sBulk liquid sSurface
dt
(17)
NO2
d [O2 ]L
.
k L a [O2 ]Sat
[O2 ]L
L
dt
(18)
20
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21
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An example of the second of these types can be found in the paper on ethanol
production by Kluyveromyces marxianus. Here metabolic production of ethanol
in an anaerobic batch reactor is described by a biochemically structured model:
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Model prediction results (smooth curves) for product formation show good
agreement with experimentally determined data (points), but as expected, do
not show such good agreement for biomass production (see run 3):
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Cell
inflow
rate
Cell
outflow
rate
Cell
birth
rate
Cell
death
rate
(19)
dC ( y, t ) C in ( y) C ( y, t )
B ( y , t ) D( y , t )
dt
where:
(20)
C = cell concentration
t = time
y = segregated function, e.g. cell age function, or distribution of different
cell functional units (spores, vegetative cells, etc.)
= reactor space time = reactor volume/inflow rate
Such bioreactors
often show
oscillatory
behaviour:
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Sin(t)
xLin(y,t)
xDin(y,t)
V
S
DO
F
S(t)
xL(y,t)
xD(y,t)
Air
dxL ( y, t ) F in
d xL ( y , t )
(21)
Rate of
change
of living cell
conc.
M.V.E. Duarte et al, Braz. J. Chem. Eng., vol.20, no.1, Jan./Mar. 2003. http://dx.doi.org/10.1590/S0104-66322003000100002
dxL ( y, t ) F in
d xL ( y , t )
(21)
where K = cell division probability density function:
2
y
y
K
K
K
K ( y, S , DO) K DO
1 DO
f DO
f SK f yK1 1 f yK2 exp P1 T 1
P2
P2
(22)
K
f DO
1 e
DO
(23)
K
f DO
DO
M.V.E. Duarte et al, Braz. J. Chem. Eng., vol.20, no.1, Jan./Mar. 2003. http://dx.doi.org/10.1590/S0104-66322003000100002
25
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K ( y, S , DO)
DO ( ppm)
M.V.E. Duarte et al, Braz. J. Chem. Eng., vol.20, no.1, Jan./Mar. 2003. http://dx.doi.org/10.1590/S0104-66322003000100002
dxL ( y, t ) F in
d xL ( y , t )
(21)
where D = cell death probability density function:
D
D
D
D( y, S , DO) 1D 2D 1 ( DO
(1 DO
) f DO
) f SD f yD1 1 f yD2
(24)
D ( y, S , DO)
DO ( ppm)
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dxL ( y, t ) F in
d xL ( y , t )
Dead cells:
dxD ( y, t ) F in
xD ( y, t ) xD ( y, t ) D( y, S , DO) xL ( y, t )
dt
V
(25)
Substrate:
S S
dS (t ) F in
S (t ) S (t ) M x
dt
V
KS S
DO DO
DO
xL ( y, t )dy
K DO DO 0
(26)
Partial differential equations such as eqs. 21, 25, and 26 may be solved by a number of
methods such as:
Finite element/method of lines/Galerkin method
Method of characteristics
Finite difference method
M.V.E. Duarte et al, Braz. J. Chem. Eng., vol.20, no.1, Jan./Mar. 2003. http://dx.doi.org/10.1590/S0104-66322003000100002
xL ( y , t )
Time (hr )
xD ( y , t )
Time (hr )
Modelling results for time variation of live and dead cell concentrations
M.V.E. Duarte et al, Braz. J. Chem. Eng., vol.20, no.1, Jan./Mar. 2003. http://dx.doi.org/10.1590/S0104-66322003000100002
27
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Cell mass ( g / L)
Time (hr )
Substrate conc., S ( g / L)
Time (hr )
Modelling results for time variation of total cell and substrate concentrations
M.V.E. Duarte et al, Braz. J. Chem. Eng., vol.20, no.1, Jan./Mar. 2003. http://dx.doi.org/10.1590/S0104-66322003000100002
dt
dm
m'
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dt
dm
m'
(m, S ' ) e m md
* 2
m mt* mo , md*
when
(28)
A exp m m' m
2
* 2
t
(29)
A, = constant parameters.
This Gaussian function has two peaks in the cell number distribution, one at m*t ,
corresponding to mother cells, and one at m*t- m corresponding to daughter cells.
k gf (G' ) k go (G' )
dG
DG f G
W (m, t )dm
dt
Ygo
Ygf
0
(30)
dG'
g G G'
dt
(31)
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k go (G' , O)
goG'
O
.
K go G' K gd O
(32)
Modelling versus experimental results for chemostat yeast fermentation oscillatory behaviour
M. A. Henson et al , Biotechnol. Prog., vol. 18, 10101026, (2002).
30
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Dilution rate
increase here
Mycoplasma genitalium
http://www.kurzweilai.net/first-complete-computer-model-of-an-organism
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3. BIOCHEMICAL MATERIAL
BALANCE
For systems at steady state (no accumulation/depletion) the total mass balance is:
Rate of input = Rate of output
of mass
of mass
(34)
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In the case of reactive species in a system at steady state, equation (33) can
also be simplified to:
0
or:
Rate of input + Rate of formation = Rate of output + Rate of consumption (35)
(where S = mass of, or number of moles of, a chemical or biochemical species)
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Assemble:
(1) Draw the flowsheet, showing all pertinent data with units.
(2) Define the system boundary and draw it on the flowsheet.
(3) Write down the reaction stoichiometric equation (if any).
B.
Analyse:
C.
Calculate:
D.
Finalise:
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35
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36
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37
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38
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39
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Taking one mole of substrate as the basis, we can write this as a balanced equation:
CwHxOyNz + aO2 + bHgOhNi
Substrate
Nitrogen
source
Primary
metabolic
product
In the chemical formula for substrate, e.g. for glucose: w=6, x=12, y=6, z=0
In the chemical formula for nitrogen source, e.g. for ammonia: g=3, h=0, i=1
The chemical formula for dry biomass is given as CHONd
Note: eq. 36 only applies to reactions involving growth and/or primary metabolic
product formation. Secondary metabolite formation requires separate stoichiometric
equations for growth and product formation.
Growth factors such as vitamins and minerals, additionally taken up in small amounts
during metabolism, are generally neglected in terms of their contribution to the
stoichiometry and energetics of the overall reaction.
Other substrates and primary products can easily be added to eq. 36, if appropriate.
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As can be seen from this table, over 90% of cell biomass can be accounted for by the
elements C, H, O and N, so cell biomass formulae are normally expressed in terms of
these elements only.
From Bioprocess Engineering Principles by Pauline M. Doran
CH1.8O0.5 N0.2
Average biomass molecular weight = 24.6 (+5-10% as residual ash/other elements)
From Bioprocess Engineering Principles by Pauline M. Doran
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04/09/2014
In order to balance eq. 36 for a particular biochemical reaction, we must determine the
stoichiometric coefficients a-f. As in balancing chemical equations, elemental balances
can be done:
Carbon:
Hydrogen:
Oxygen:
Nitrogen:
w = c + d + fj
x + bg = c + 2e + fk
y + 2a + bh = c +2d + e + fl
z + bi = cd + fm
(37)
(38)
(39)
(40)
However we have six unknowns (a-f) and only four simultaneous equations, so these
cannot be solved for a-f. In addition, the fact that water is usually present in great
excess in biochemical reactions, often leads to difficulties in experimentally quantifying
changes in water concentration. This in turn means that H and O balances can be
unreliable.
Alternative stoichiometric information can be obtained by using an experimentally
determined respiratory quotient (RQ) for the reaction of interest:
RQ
moles O2 consumed
a
or aRQ = d
(41)
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Applying this idea (i.e. LHS = RHS) to eq. (36), for the case where there is no
product formation (cell growth only):
CwHxOyNz + aO2 + bHgOhNi
gives:
wS 4a = cB
(36a)
(42)
YXS
(43)
Many factors can affect the value of a yield coefficient including nature of C and N
sources, pH, temperature, and in aerobic cultures nature of the oxidising agent.
However, when the yield coefficient is constant, its experimentally measured value can
be used to determine c in equations (36) or (36a), since eq. (43) can be written in
stoichiometric terms (assuming 1 mole substrate as the basis) as:
YXS
c ( MW biomass)
( MW substrate)
(44)
Care must be exercised when using eq. (44), since it does not apply if a significant
amount of substrate is used for maintenance activities instead of growth. In such cases
the experimentally measured values of YXS must be adjusted to account for this.
We can also have a yield coefficient for primary metabolic product from substrate
YPS
(45)
Again we must remember that eq. (45) only applies for primary metabolic product.
3.2.3 Metabolic stoichiometric calculations: summary
From our considerations in the previous two sections, we can see that it is now
possible to balance our stoichiometric metabolism equation (36) for all 6 unknowns:
CwHxOyNz + aO2 + bHgOhNi
cCHONd + dCO2 + eH2O + fCjHkOlNm (36)
Carbon balance:
Nitrogen balance:
Respiratory quotient:
Electron balance:
Yield coefficient biomass:
Yield coefficient product:
w = c + d + fj
z + bi = cd + fm
aRQ = d
wS 4a = cB
c = YXS (MW substrate)/(MW biomass)
f = YXP (MW substrate)/(MW product)
(37)
(40)
(41)
(42)
(44)
(45)
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Assumptions: reaction occurs only in the chemostat; separation occurs only in separator; no
biomass in the feed or product streams (only in the cell waste stream). Let r X = biomass growth
rate and rS = substrate consumption rate.
Accumulation/depletion = Input
dX
dX R
VR 1 VS
0
dt
dt
Output
Reaction
FW X R
rX VR
At steady state:
0
FW XR
rXVR
Thus wastage rate of biomass must equal growth rate of biomass to maintain s.s.
Assumptions: reaction occurs only in the chemostat; separation occurs only in separator; no
biomass in the feed or product streams (only in the cell waste stream). Let r X = biomass growth
rate and rS = substrate consumption rate.
Output
F1S P FW S R
F1SP - FW SR
Reaction
rsVR
+ rSVR
Here the consumption rate of S equals the difference between the inlet and outlet mass
flow rates of S.
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Material balances around individual units are often more useful for quantifying dynamic
(non-steady state) behaviour.
Biomass balance around the chemostat:
VR
dX 1
dt
FR X R
( F FR ) X 1
rX VR
dS1
dt
F S0 FR S R
( F FR ) S1
rsVR
dX R
dt
( F FR ) X 1 ( FW FR ) X R
dS S
dt
( F FR ) S1
( FW FR ) S R F1S P
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Other important flow rate equations necessary to solve material balances with recycle:
Recycle flow rate:
FR
R.F
(F+FR)/C
Values of R and C must be chosen so that the cell waste flow rate (F W ) is positive.
dX 1
VR
dt
dX 1
dt
( F FR ) X 1
FR X R
[ FR X R
( F FR ) X 1
rX VR
rX VR ] / VR
dS
VR 1
dt
dS1
dt
F S0 FR S R
[ F S0 FR S R
( F FR ) S1
( F FR ) S1
rsVR
rsVR ] / VR
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At t = 60 min
X1 (biomass in Reactor) = ~8
S1 (Substrate in the Reactor = 0
XR (Biomass in Recycle Line) = ~25
SS (Substrate in Settler = 0
All substrate removed at ~ t = 20- mins
At t = 60 min
X1 (biomass in Reactor) = ~8
S1 (Substrate in the Reactor = 0
XR (Biomass in Recycle Line) = ~30
SS (Substrate in Settler = 0
All substrate removed at ~ t = 5 mins
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At t = 60 min
X1 (biomass in Reactor) = 0
S1 (Substrate in the Reactor = 0
XR (Biomass in Recycle Line) = 0
SS (Substrate in Settler = 10
At t = 60 min
X1 (biomass in Reactor) = ~8
S1 (Substrate in the Reactor = 0
XR (Biomass in Recycle Line) = ~30
SS (Substrate in Settler = 0
All substrate removed at ~ t = 18 mins
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At t = 60 min
X1 (biomass in Reactor) = ~8
S1 (Substrate in the Reactor = 0
XR (Biomass in Recycle Line) = ~38
SS (Substrate in Settler = 0
All substrate removed at ~ t = 22 mins
At t = 60 min
X1 (biomass in Reactor) = ~8
S1 (Substrate in the Reactor = 0
XR (Biomass in Recycle Line) = ~12
SS (Substrate in Settler) = 0
All substrate removed at ~ t = 25 mins
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4. MASS TRANSFER
Doran (2nd Edition), Chapter 13.
4. Mass Transfer
Due to the complex nature and rheological behaviour of many biochemical systems, a
detailed understanding of mass transfer is often important in order to be able to quantify
biochemical processing operations.
There are two major aspects to mass transfer in biochemical processes: internal and
external.
Internal mass transfer is concerned with the movement of substrates and products
within cellular agglomerates or immobilised enzyme systems, and is of major
importance in quantifying reaction rates.
External mass transfer deals with the transport of materials in the fluid phase(s)
surrounding the biochemically reactive entity, and features prominently in both
reactions (particularly in aerobic and/or highly viscous reaction media), and in
separation processes.
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Int. m.t.
Ext. m.t.
R s
a s
max
s
s
(14)
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is related to the tortuosity factor (Thiele modulus), , of the gel/medium, for example
for 1st order reaction kinetics in a flat plate:
tanh
Floc,
film,
or particle
CAs
Concentration of A at
particle surface = CAs
NA
dC A
k s a C Ab C As
dt
(17)
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Liquid
film
Bulk liquid
(substrate
solution)
Oxygen concentrations at
the gas-liquid interface
Gas
film
Bulk gas
(bubble)
[O2]L(i)
[O2]
[O2]L
Gas-liquid
interface
NO2
d [O2 ]L
.
k L a [O2 ]Sat
[O2 ]L
L
dt
(18)
JA
NA
dC A
DAB
a
dy
(47)
JA = mass flux of A
NA = rate of mass transfer of A
a = mass transfer area
DAB = diffusivity of A in a mixture of A and B
Concentration gradient of component A
inducing mass transfer across area a
From Bioprocess Engineering Principles by Pauline M. Doran
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Accum.
/dissip.
of A
0
Input
of A
= DAe
Output
of A
dC A
dC A
- DAe
4 r 2
4 r 2
dr
dr
r
r r
Formation/ (33)
consumption
of A
rA 4 r 2 r
(CA = concn. of A)
Dividing by 4r gives:
DAe dC A r 2
DAe dC A r 2
dr
dr
r r
r
rA r 2
r
or
dC A 2
DAe
r
dr
r r2
A
r
where the numerator term refers to the difference in the two numerator terms from
the previous equation.
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dC A 2
d DAe
r
dr
r r2
A
dr
dC
d A r2
dr
r r2
DAe
A
dr
The bracketed term is the derivative of a product, d(u.v)/dx, where u = dCA/dr and
v = r2. Thus we can expand this derivative to give:
d 2C A 2
dC
DAe
r 2r A
2
dr
dr
rA r 2
(48)
Integration of this 2nd order differential equation yields an expression for how C A varies
with distance (r) inside the particle. This must be done on a case-by-case basis
according to the reaction kinetics however, since rA is a function of CA in most cases.
4.2.3 Substrate concentration profile: 1st order kinetics & spherical geometry
d 2C
dC
A 2
With 1st order kinetics, eq. (48) becomes: DAe
r 2r A k1C A r 2 0
2
st
dr
dr
(k1 = 1 order rate constant)
(49)
According to assumptions (i), (iii) and (vi) above, k 1 and DAe can be considered
constant. Since this is a 2nd order differential equation, two boundary conditions are
required:
dC A
CA = CAs at r = R, and
0 at r = 0
dr
where CAs is substrate concentration at the particle surface. The latter boundary
condition is called the symmetry condition: the substrate concentration profile is
symmetrical at the centre of the particle, thus the slope (dC A/dr) is zero at r=0.
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C A C As
R sinh r k1 / DAe
r sinh R k1 / DAe
(50) ,
where
sinh( x)
e x e x
2
Eq. (50) can be used to calculate the particle substrate concentration profile.
4.2.4 Substrate concentration profile: zero order kinetics & spherical geometry
In this case eq. (48) becomes:
(k0 = zero order rate constant)
d 2C A 2
dC
DAe
r 2r A
2
dr
dr
k0 r 2 0
(51)
Assuming that CA can be zero only at r = 0 (centre of the sphere), integration of eq. (51)
with the same boundary conditions as before, gives:
C A C As
k0
r 2 R2
6 DAe
(52)
It is important, from a practical perspective, that the particle core does not become
starved of substrate. This is more likely with larger particles. Here we can calculate
the maximum particle size, Rmax where CA > 0 (depletion only occurs at r = 0) from
eq. (51), since then CA = r = 0, and:
6 DAe C As
(53)
Rmax
k0
d 2C
dC
A 2
In this case eq. (48) becomes: DAe
r 2r A max A r 2 0
2
dr K m C A
dr
(54)
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and
CA = CAs
at
z = b,
dC A
dz
at
z=0
Substrate concentration
profile in an infinite flat plate
From Bioprocess Engineering Principles by Pauline M. Doran
(50)
(55)
(52)
(56)
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rA,Obs
rR
r 0
k1C A dV p
(57)
e x e x
e x e x
Zero order reaction: Assuming CA > 0 everywhere in the particle, then the rate will be
constant and independent of CA. Thus the overall rate is simply the rate constant
multiplied by the particle volume:
rA,Obs
4
R 3 k0
3
(59)
4
rAs R 3k1C As
3
(61),
i ,1
3 DAe
R k1 / DAe coth R k1 / DAe 1
R 2 k1
(62)
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V p rAs C As
C DAe rA dC A
S x 2 A,eq
(63)
where: Vp = particle volume, Sx = external surface area, CA,eq = equilibrium [A] (=0 for
most biochemical reactions), and rA = reaction rate. From geometry, Vp/Sx = R/3 for
spheres, and = b for flat plates.
For 1st order kinetics with spherical geometry, we have:
and:
1
(65)
31 coth 31 1
i ,1
R
3
k1
DAe
(64)
312
and i for other kinetics and geometries can be quantified by similar equations or
numerical methods.
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Vp
Sx
rA,Obs
DAe C As
(66)
R rA,Obs
Sphere
3 DAe C As
Flat plate b 2
rA,Obs
DAe C As
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dC A
k s a C Ab C As
dt
(17)
Sh
Schmidt number:
Sc
ks Dp
L
L DAL
(67)
DAL
(68)
D p u pL L
L
gD L ( p L )
Gr
L2
3
p
(69)
(70)
Sh
ks Dp
DAL
Sc
L DAL
Re p
D p u pL L
Gr
gD 3p L ( p L )
L2
Sh:
Sc:
ReP:
Gr:
The form of the correlation(s) used to estimate k s depends on the configuration of the
mass transfer system, the flow conditions and other factors. In all cases, ultimately the
Sherwood number, Sh, must be evaluated.
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Sh
ks Dp
DAL
Sc
L DAL
Re p
D p u pL L
Gr
gD 3p L ( p L )
L2
Free-moving spherical particles. For this situation, the rate of mass transfer depends
on the slip velocity, upL, which is difficult to measure and must be estimated before
calculating ks from Sh.
1. Calculate Gr:
For Gr<36
Re p
Gr
18
(71)
For 36<Gr<8x104
Re p 0.153Gr 0.71
(72)
For 8x104<Gr<3x109
Re p 1.74Gr 0.5
(73)
Sh 4 1.21Re p Sc
(74)
(75)
0.67
Sh
ks Dp
DAL
Sc
L DAL
Re p
D p u pL L
Gr
gD 3p L ( p L )
L2
Spherical particles in a packed bed. Here, ks depends on the liquid linear velocity, u,
around the particles. This is measureable and replaces the slip velocity u pL, in the
calculation of Rep.
1. Calculate Rep: For 10<Rep<104
(76)
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Sx
C Ab C As
Vp
V p rA,Obs
C As
1
C Ab
S x k s C Ab
, or
(77)
V p rA,Obs
(78)
S x k s C Ab
All of the rhs terms are usually measurable. To assess whether or not external diffusion
is important in determining the overall rate determining step, apply the following criteria:
If << 1, then CAs CAb, and external mass transfer is not rate limiting.
If is significant (typically greater then 0.05), then CAs < CAb, and external
mass transfer is limiting.
For typical particle geometries we can write:
R rA,Obs
Sphere
3 k s C Ab
Flat plate b
rA,Obs
k s C Ab
rA,Obs
rAb
observed rate
rate if C A C Ab throughout the particle
(79)
T A,Obs As i e
rAs rAb
(80)
(81)
C As
C Ab
C
C K C Ab
Michaelis Menten rA max A e, M As m
K
C
C Ab K m C As
m
A
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V p rA,Obs
(66)
S x DAe C As
The observable Thiele modulus, , indicates that internal mass transfer rate can be
improved by:
Internal mass transfer
Decreasing particle size. This has the biggest effect on , since the latter is
proportional to the square of particle size (R2 for spheres and b2 for plates).
There may be practical limitations however on this, due to poorer mechanical
strength and increased difficulty in retention of the particles within the reactor
system.
V p rA,Obs
S x k s C Ab
(78)
The observable external mass transfer modulus, , indicates that external mass
transfer rate can be improved by:
Decreasing the particle size. This increases the external particle surface area
and reduces the thickness of the boundary layer.
Increasing the mass transfer coefficient, ks. This may be best done by
increasing the liquid (linear) velocity adjacent to the particle surface. Changing
the liquid viscosity and density, or increasing the substrate diffusivity in the liquid
will also increase ks.
Increasing the substrate bulk concentration. This increases the driving force for
external mass transfer according to eq. (17)
Reducing the observed reaction rate, rA,Obs. See similar comments on this,
under improving the internal mass transfer rate.
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5. BIOCHEMICAL ENERGY
BALANCES
Energy in/out by
shaft work, Ws
Energy of
in-flowing
materials
The bioprocessing
system
Energy of
out-flowing
materials
Energy in/out by
heat transfer, Q
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(79)
Referring to the various energy transfer modes possible, we can quantify eq. (79) to
obtain a series of general energy balance equations:
E = Mi (Ek + Ep + U + pV)i - Mo (Ek + Ep + U + pV)o - Q + W s
(80)
(81)
(82)
In the case (true for many biochemical reaction systems) where there is little change in
kinetic or potential energy, and where the system is operating at steady state (no
energy accumulation/depletion), we can further simplify (82) to give:
Mi hi - Mo ho - Q + W s
= 0
or, in cases where there may be more than one input and one output streams:
= 0
(83)
(84)
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Temperature changes
Change of phase e.g. liquid to gas
Mixing or dissolution
Reaction
(85)
M = mass
Cp = heat capacity at constant pressure.
The heat capacity value for a given substance can vary with temperature. These are
often given in the literature as a polynomial function of temperature, such as :
Cp,i i i T i T 2
(86)
(87)
Since large temperature changes do not often occur in bioprocessing, in many cases
Cps are assumed constant.
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H = M. hv
(88)
where:
Whereas H0f,i values are often available for chemical components, this is often not
the case for biochemicals.
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Instead heats of combustion, h0c,i are used. These are relatively easily measured
since all biochemical materials are combustible, giving simple gases such as CO 2, H2O
vapour and N2. Thus:
(90)
Equation (90) allows calculation of the standard enthalpy of reaction (i.e. normally at
25oC). For reactions carried out at other temperatures, then sensible heat changes
have also to be taken into account, as outlined in section 5.2.1. For most biochemical
reactions, this is not an issue. However one major exception is in the case of single
enzyme conversion reactions. These have very small reaction enthalpy changes, and
hence any additional sensible heat effects can be quite significant in determining the
overall enthalpy change.
The enthalpy of reaction at non-standard conditions can be quantified by considering
the hypothetical path that involves the same initial and final states of the reaction
mixture, but takes place via the standard reaction conditions.
Actual path
0
H rxn (at T ) H1 H rxn
H 3
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Thus either equation (91) (for aerobic fermentations) or equation (90) (for all
fermentation types) can be used to calculate Hr if either the fermentation oxygen
consumption or the reaction component heats of combustion are known.
Heat of combustion of biomass have been found experimentally to fall into two
broad groups:
Bacteria
Yeasts
= 0
(83)
it is possible to formulate a useful version for this particular case (cell bioreactors):
-Hr
Mv.hv - Q + W s = 0
(92)
since reaction and latent heat are the only significant contributors to enthalpy change
between the inlet steams and the outlet streams.
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5.2.6 Essential good practice and procedure for performing energy balance
calculations
These are essentially the same as for material balances (section 3.1).
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78