1

ELEMENTALAND REDOX
BALANCES
(2 LECTURES)
Chapter 3 VNL
Content of lectures
Fundamental balances in CSTR
Nomenclature & concepts
Batch, Fed-batch, Chemostat
Steady-state & transients
Rates
Volumetric and specific
The C-mole concept
Examples of compounds
Examples of reactions
Biomass?
Black box balances
Yield factors (C-mol yield vs. mass yield)
Carbon balances
Degree of reduction balances
2
Mass balances in reactors
Typical lab-scale set-up
Substrate reservoirs
Acid Base
Substrate
Effluent liquid
Gas feed
Exhaust gas
Liquid feed
Balance
Substrate reservoirs
Acid Base
Substrate
Effluent liquid
Gas feed
Exhaust gas
Liquid feed
Balance
Mode of operation
Batch
Continuous operation
Fed-batch
3
Mass balances
Accumulation = Inflow Outflow + Produced Consumed
The accumulation term = 0 at steady-state
Chemostat
( ) 0
,
= +
i f i s
s s v V q
i
( ) 0
,
= +
i f i p
p p v V q
i
( ) 0 = + x x v V q
f x
V
v
D = = rate Dilution
4
Nomenclature
q = volumetric rates (C-mole/l h)
r = specific rates (C-mole/C-mole h)
Y
ji
= yield coefficients (C-mole /C-mole)
s = substrate conc. (C-mole /l)
x = biomass conc.
p = product conc.
Macroscopic (black box description
Intracellular
biochemical
reactions
Substrates
Metabolites
Extracellular
macromolecules
Biomass Mainly:
Carbon source (e.g. glucose)
nitrogen source (e.g. ammonia)
5
The concept of C-moles
0 O H .... CO .... NH O O CH
2 1 2 1 3 2 2
1 1
= + + + + +
sw sp sc sx ss sn so
Y P Y Y X Y S Y Y Y
glucose
Ammonia (nitrogen source)
oxygen
Biomass
Carbon dioxide
Metabolic product
Microorganism Elemental
composition
Ash
content
(w/w %)
Condition
Candida utilis
Klebsiella aerogenes
Saccharomyces cerevisiae
Escherichia coli
Pseudomonas fluorescens
Aerobacter aerogenes
Penicillium chrysogenum
Aspergillus niger
Average
CH
1.83
O
0.46
N
0.19
CH
1.87
O
0.56
N
0.20
CH
1.83
O
0.54
N
0.10
CH
1.87
O
0.56
N
0.20
CH
1.75
O
0.43
N
0.22
CH
1.73
O
0.43
N
0.24
CH
1.75
O
0.47
N
0.17
CH
1.73
O
0.43
N
0.24
CH
1.82
O
0.58
N
0.16
CH
1.78
O
0.60
N
0.19
CH
1.94
O
0.52
N
0.25
P
0.025
CH
1.77
O
0.49
N
0.24
P
0.017
CH
1.83
O
0.50
N
0.22
P
0.021
CH
1.96
O
0.55
N
0.25
P
0.022
CH
1.93
O
0.55
N
0.25
P
0.021
CH
1.83
O
0.55
N
0.26
P
0.024
CH
1.64
O
0.52
N
0.16
CH
1.72
O
0.55
N
0.17
CH
1.81
O
0.52
N
0.21
7.0
7.0
7.0
7.0
3.6
3.6
3.6
3.6
7.3
9.7
5.5
5.5
5.5
5.5
5.5
5.5
7.9
7.5
6.0
Glucose limited, D=0.05 h
-1
Glucose limited, D=0.45 h
-1
Ammonia limited, D=0.05 h
-1
Ammonia limited, D=0.45 h
-1
Glycerol limited, D=0.10 h
-1
Glycerol limited, D=0.85 h
-1
Ammonia limited, D=0.10 h
-1
Ammonia limited, D=0.85 h
-1
Glucose limited, D=0.080 h
-1
Glucose limited, D=0.255 h
-1
Unlimited growth
Unlimited growth
Unlimited growth
Unlimited growth
Unlimited growth
Unlimited growth
Unlimited growth
Unlimited growth
X~ CH
1.8
O
0.5
N
0.2
6
What is biomass?
Macromolecule Elemental
composition
Percent by
weight
g (C-mole)
-1
Protein
RNA
DNA
Carbohydrates
Phospholipids
Neutral fat
Pool of cellular metabolites
CH
1.58
O
0.31
N
0.27
S
0.004
CH
1.25
O
0.25
N
0.38
P
0.11
CH
1.15
O
0.62
N
0.39
P
0.10
CH
1.67
O
0.83
CH
1.91
O
0.23
N
0.02
P
0.02
CH
1.84
O
0.12
CH
1.8
O
0.8
N
0.2
S
0.01
57
16
3
10
10.8
2.5
0.7
22.45
34.0
31.6
27.0
18.5
15.8
29.7
Example S. cerevisiae
Yield coefficients
1
and

= =
ji ij
j
i
j
i
ji
Y Y
q
q
r
r
Y
7
Fig. 3.4
0
40
80
120
160
0 100 200 300 400
- q
glc
(C-moles h
-1
)
(
C
-
m
o
l
e
s

h
-
1
)
ethanol
CO
2
biomass
glycerol
V
0
40
80
120
160
0 100 200 300 400
- q
glc
(C-moles h
-1
) (C-moles h
-1
)
(
C
-
m
o
l
e
s

h
-
1
)
ethanol
CO
2
biomass
glycerol
V
Degree of reduction
Define a redox neutral compound for each element of
interest.
We choose H
2
O, CO
2
, NH
3
, H
2
SO
4
, H
3
PO
4
as the
neutral compounds corresponding to the elements O,
C, N, S and P. With this set of neutral compounds
and with the unit of redox defined as H = 1 one
obtains the following redox levels of the five listed
elements:
O = -2 , C = 4 , N = -3 , S = 6 , P = 5
8
Degree of reduction
Now the redox level of any reactant in a biochemical
reaction can be calculated. With our convention to
write all carbon containing compounds on a 1 C-atom
basis one obtains the following degrees of reduction
k
i
.
CH
2
O (glucose and other hexoses, acetic acid, lactic
acid, formaldehyde), k = 4.
CH
3
O

(ethanol), k = 6.
CH
2
O
2
(formic acid), k = 2.
CH
1.8
O
0.5
N
0.2
(standard biomass), k = 4.20
Degree of reduction
(alternative derivation)
glucose + ammonia + oxygen biomass + carbon dioxide + water
Consider the base case of aerobic growth on glucose and ammonia, i.e.:
One can set up an elemental balance for C, H, O and N on matrix form
according to:
Eq = 0
where E is a 4 x N matrix, i.e. a matrix in which the i-th column
gives the elemental composition of compound i (there are N compounds),
and q is the vector of net formation rates of the N compounds.
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For a standard biomass composition of CH
a
O
b
N
c
we get:
(
(
(
(

=
(
(
(
(
(
(
(
(

(
(
(
(

0
0
0
0
0 1 0 0 0
2 0 1 2 1
0 3 2 0 2
0 0 1 0 1 1
o
n
x
w
c
s
q
q
q
q
q
q
c
b
a
glucose
biomass
carbon dioxide
water
ammonia
oxygen
C
H
O
N
The four elemental balances will thus be:
0 = + +
x c s
q q q

0 3 2 2 = + + +
n x w s
q aq q q

0 2 2 = + + + +
o x w c s
q bq q q q

0 = +
n x
q cq
(C)
(H)
(O)
(N)
Mathematics, in its generosity, allows us to multiply each equation with a
freely chosen constant (= 0!). If we choose one constant for each equation,
denoted k
C,,
k
H
, k
O
and k
N
add the four equations and rearrange the terms,
we get:
( ) ( ) ( ) ( )
( ) 0 3
2 2 2
= + +
+ + + + + + + + + + +
o O n N H
x N O H C w O H c O C s O H C
q q
q c b a q q q
k k k
k k k k k k k k k k k
10
Now we can choose the multipliers (the k) in such a way that the coefficients
in front of q
c
, q
w
and q
n
vanish. This requires that
0 2 = +
O C
k k

0 2 = +
o O H
k k

0 3 = +
N H
k k
If we finally choose (as normalization)
1 =
H
k we arrive at the values
4 =
C
k
2 =
O
k
3 =
N
k
Why make carbon and degree of
reduction balances?
Some of the products may go undetected because it
was not suspected that they would be formed in the
bioreaction.
Products that were thought to remain in the liquid
phase are in fact partly stripped to the gas phase.
The instruments used to measure the rates are
wrongly calibrated or they malfunction after some
time.
11
Important
Concepts such as:
Chemostat
Dilution rate
Yield coefficients
C-mole
RQ = respiratory quotient
degree of reduction
Study the examples!