MASS & ENERGY

BALANCE
Case study:
Production of citric acid by
Aspergillus niger using cane
molasses in a fermentor
Electronic Journal of Biotechnology ISSN: 0717-3458 Vol.5
No.3, Issue of December 15, 2002 2002 by Universidad
Catlica de Valparaso -- Chile Received May 16, 2002 /
Accepted November 5, 2002

A laboratory scale stirred fermentor of 15-L capacity having

working volume of 9-L was used for cultivation process and
nutritional analysis. The strain GCBT7 Aspergillus niger, was
found to enhance citric acid production.

Volume of fermenter: 15 L
Working volume: 9 L
pH value = pH 6.0
Incubation temperature = 30C
Raw molasses sugar-mainly sucrose (Substrate): 150
g/L
Fermentation hours: 144 hours.
Ammonium nitrate (Nitrogen source) = 0.2%= 2g/L
Maximum Production citric acid (Product) : 99.56%
3.5 g/L
The dry cell mass, A.niger (Biomass): 18.5 g/L
(Notes: Assume 100% consumption of sugar and N
source).
Yields: 1) Yx/s (Biomass yield from substrate) =
0.123
2) Yp/s (Product yield from substrate) =
0.664

MASS BALANCE
1) Develop the product stoichiometric
equation
-assume extracellular product (citric acid)

Substrate (Carbon source-sucrose from molasses)Biomass

(A.niger)

CwHxOyNz + aO2 + bHgOhNi

cCHON + dCO2 +
eH
2O + fCjHkOlNm
Nitrogen source (Ammonium nitrate)
Product (citric acid)

CwHxOyNz + aO2 + bHgOhNi

cCHON + dCO2 + eH2O +
fCjHkOlNm
Stoichiometric coefficient balance:

C balance: w = c + d + fj
H balance: x + bg = c + 2e
+ fk
O balance: y + 2a + bh = c
+2d +e + fl
N balance: z + bi = c + fm

C12H22O11 + aO2 + bNH4NO3

cCH1.72O0.55N0.17 +
dCO2 + eH2O + fC6H8O7
2) Calculate the stoichiometric coefficient balance
C balance : 12 = c + d + 6f
H balance: 22 + 4b = 1.72c + 2e + 8f
O balance: 11 + 2a +3b = 0.55c + 2d + e + 7f
N balance: 2b = 0.17c

x/s

= c (Mw cells) / (Mw substrate) **

where Yx/s = 0.123

Mw cells = 24.9 + ash (7.5%)
= 24.9 / (1 0.075)
= 26.92 g/mol
Mw substrate (sucrose) = 342 g/mol
0.123 = c (26.92 / 342)
c = 1.56

N balance: 2b = 0.17c
b = (1.56 * 0.17) /2
b = 0.133
Yp/s = 0.664
Yp/s = f (Mw product) / (Mw substrate)
**
Mw citric acid = 192 g/mol
Mw sucrose = 342 g/mol
f = 1.15

C balance : 12 = c + d + 6f
d = 12 -1.56 6(1.15)
d = 3.54
H balance: 22 + 4b = 1.72c + 2e + 8f
22 + 4(0.133) = 1.72(1.56) + 2e +
8(1.15)
e = (22.944 15.58) / 2
e = 5.32

O balance: 11 + 2a +3b = 0.55c + 2d

+ e + 7f
a = (16.899 - 11.708) / 2
a = 4.95

C12H22O11 + 4.95O2 + 0.133 NH4NO3

1.56 CH1.72O0.55N0.17 + 3.54CO2 +
5.32H2O +
1.15 C6H8O7

C12H22O11 + 4.95O2 + 0.133 NH4NO3

1.56CH1.72O0.55N0.17 + 3.54CO2 + 5.32H2O
+
1.15 C6H8O7
1
1
1
1

mol
mol
mol
mol

C12H22O11
C12H22O11
C12H22O11
C12H22O11

produces
produces
produces
produces

1.15
1.56
3.54
5.32

mol
mol
mol
mol

C6H8O7
CH1.72O0.55N0.17
CO2
H2O

Estimation of plant capacity: 100 tonnes citric ac

Mol of 100 tonnes of citric acid = 1 x 108 g / Mw citric
acid
= 1 x 108 g / 192 g/mol
= 520 833 moles

Amount of sucrose consumed:

= (mol sucrose / mol citric acid) * total
no. of mol citric acid) * Mw sucrose
= (1 / 1.15) * 520 833 moles * 342 g/mol
= 154.89 tonnes sucrose/year
Amount of O2 consumed:
= (4.95/1.15) * 520 833 moles * 32 g/mol
= 71.74 tonnes O2 /year
Since the biomass is
also a product side,
so, use sucrose as
basis i.e.,
Mol of 154.89 tonnes
of sucrose = 1.55 x
108 g / Mw sucrose
= 1 x 108 g / 342
g/mol
= 452895 moles)

Amount of biomass produced:

= (1.56/1) * 452 895 moles * 26.92 g/mol
= 19.02 tonnes biomass / year

Amount of Ammonium nitrate consumed:

Mw NH4NO3 = 80 g / mol
= (0.133/1.15) * 520 833 moles * 80 g/mol
= 4.82 tonnes NH4NO3 / year
Amount of CO2 produced:
Mw CO2 = 44 g / mol
=(3.54/1) * 452 895 moles * 44 g/mol
= 70.54 tonnes CO2 / year
Amount of H2O produced:
Mw H2O = 18 g / mol
=(5.32/1) * 452 895 moles * 18 g/mol
= 43.37 tonnes H2O/ year

Mass Balance

Estimation of plant capacity: 100 tonnes citric a

Off gas

Molasses
(1032.60
tonnes/year):
15%
sucrose=154.89
tonnes/year
85% H2O
in=877.71
Ammonium
tonnes/year
nitrate:
0.2 % (2g/L) = 4.82
tonnes/year

(340.42
tonnes/year):
CO2= 70.54
tonnes/year
N2= 269.88
tonnes/year

Fermenter

Air
(341.62 tonnes/year):
21%O2= 71.74
tonnes/year
79% N2= 269.88
tonnes/year

Biomass (A.niger):
19.02 tonnes/year
Citric acid:
100 tonnes/year
H2O out
(921.08
tonnes/year):
H2O produced=
43.37 tonnes/year
H2O in=877.71
tonnes/year

Total Mass Balances (MASSin MASSout):

Stream

Mass In
(tonnes/year)

Mass Out
(tonnes/year)

Sucrose

154.89

4.82

O2

71.74

N2

269.88

269.88

Biomass, A.niger

19.02

Citric acid

100.00

CO2

70.54

877.71

921.08

1379.04

1380.52 *

Ammonium
nitrate

Water

Total

Note: * some portions of water lost due to evaporation

ENERGY BALANCE
Considering the various quantities of materials
involved, their specific heats, and their changes
in temperature or state.
Heat Management in Bioreactors:
Temperature control essential for optimisation of
biomass production or product formation.
Typical cultivation conditions include:
Small reactors large surface area to unit
volume ratio generally require heat addition.
Large reactors small surface area to unit
volume ratio generally require heat removal.

General operating temperature of microbes

Growth Temp.
(0C)
Species

Min.

Opt.

Max.

Plant cells

---

25

---

Animal cells

---

37

---

E. Coli

10

30-37

45

B. Subtilis

15

30-37

55

S. Cerevisiae

0-5

28-36

40-42

Heat Balancing
General energy balance can be applied to a bioreactor.

Qacc= Qmet + Qag + Qaer + Qsen - Qevap - Qhxcr - Qsurr

Where
Qacc is the accumulated energy in the system (can be
positive or negative in the case of heat loss)
Qmet Energy generated by metabolism
Qag Energy generated by agitation (W)
Qaer Energy generated by aeration (W)
Qsen Energy generated by condensation (sensible heat)
Qevap Heat loss to evaporation
Qhxcr Heat loss to heat exchanger (can be negative or
positive)
Qsurr Heat loss to surrounding environment

We require steady state conditions in a

fermenter, there in fermentation we
require Qacc=0.
If we ignore heat loss to the
surrounding environment (usually
negligible) we can describe the heat
exchanger duty as:
Qhxcr = Qmet + Qag + Qaer + Qsen Qevap

Energy Balance (study case)

Assumption:
-no shaft work (impeller), W s=0 (in this example)
-no evaporation, Mv=0
-heat of reaction, Hc at 30 C is -460 kJ gmol-1 O 2 consumed
(for aerobic-consider only O2 combustion,
for anaerobic, you have to find Hc for each of the reactants &
products).
- Q accumulation, Qacc = 0
- Negligible sensible Heat change, Q sen = 0

Energy balance equation:

For cell metabolism, the modified energy balance equation is:

Hrxn Mvhv Q Ws = 0
In this case, since Ws= 0; Mv= 0, therefore:

Hrxn Q = 0

Hrxn is related to the amount of oxygen

consumed:
Hrxn = (-460 kJ gmol-1) * (71740 kg) *
Data
mass balance
(1000g /1kg)
* (1from
gmol/
32 g)
= -1.03 x 1010 kJ
Since;
Hrxn Q = 0
Q = +1.03 x 1010 kJ / year
(amount of heat that must be removed from the
fermenter per 100 tonnes citric acid produced )

Energy Balance

Estimation of plant capacity: 100 tonnes citric aci

Off gas

Molasses
(1032.60
tonnes/year):
15%
sucrose=154.89
tonnes/year
85% H2O
in=877.71
Ammonium
tonnes/year
nitrate:
0.2 % = 4.82
tonnes/year

(340.42
tonnes/year):
CO2= 70.54
tonnes/year
N2= 269.88
tonnes/year

Fermenter
30 C

Air
(341.62 tonnes/year):
21%O2= 71.74
tonnes/year
79% N2= 269.88
tonnes/year

Q= +1.03 x
1010 kJ

Biomass (A.niger):
19.02 tonnes/year
Citric acid:
100 tonnes/year
H2O out
(921.08
tonnes/year):
H2O produced=
43.37 tonnes/year
H2O in=877.71
tonnes/year

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