UNIT 8

STOICHIOMETRY OF BIOPROCESS
The growth of biomass with time is given by
substrate + cells extracellular products + more cells

The rate of microbial growth is characterized by specific growth rate

net =

1 dX
X dt

net = specific growth rate

X = cell concentration
t = time
net = g - kd where, g= gross specific growth rate,
kd= death rate of cells , if kd =0, net = g
Monod eq: applied for balanced growth, i.e. composition of biomass remains constant and
specific rate of production of each component of culture is equal to .
rZ = z, where Z cellular component(ex: protein, RNA, polysaccharide), rZ is volumetric rate of
production of Z, z is concentration of Z
Where m = max specific growth rate,
when S>>Ks, g = m
when S<<Ks g = m S/ KS

Limitations of Monods eq:

applied only for balanced growth

At extremely low substrate concentration cannot be applied

YIELD

True or stoichiometric or theoretical yield = (total mass or mole of product formed)

/(mass or moles of reactant used to form that particular product)

Observed or apparent yield = (mass or moles of product present) /(total mass or moles of
reactant consumed)

YIELD COEFFICIENT

YX/S = mass or moles of biomass produced per unit mass or moles of substrate
consumed

YP/S = mass or moles of product formed per unit mass or moles of substrate consumed

YP/X = mass or moles of product formed per unit mass or moles of biomass
biom formed

YX/O = mass or moles of biomass formed per unit mass or moles of oxygen consumed

YCO2/S = mass or moles of CO2 formed per unit mass or moles of substrate consumed

RQ = moles of CO2 formed per mole of O2 consumed

YATP = mass or moles of bioma

biomass
ss formed per unit mass or moles of ATP formed

Ykcal = mass or moles of biomass formed per Kcal of heat evolved in fermentation

Maintenance coefficient : specific rate of substrate uptake for cellular cell activities.
m =

(ds / dt ) m
X

Types of maintenance coefficients:

1
1
m
=
+ ATP (1)
AP
M
D
YX
/ ATP YX / ATP
m
1
1
O2
=
+
(2)
AP
M
D
Y
Y
X / O2
X / O2
D= dilution factor
ELEMENTAL BALANCES (only biomass is produced without product formation)

C balance: w = c+d

H balance: x+bg = c+2e

O balance: y+2a+bh = c+2d+e
N balance: z+bi=c
RQ=d/a
5 eqs. and 5 unknowns (a,b,c,d,e) solving gives 5 unknowns
Degree of reduction (): Number of equivalents of available electrons per gram atom Carbon.
Available electrons are those that would be transferred to oxygen upon oxidation of a compound
to CO2, H2O, NH3.
for C =4, H=1, N=-3, O=-2, P=5, S=6
Ex: For CH4: [1(4)+4(1)] /1 =8
C6H12O6: [6(4)+12(1)+6(-2)] / 6 =24/6 = 4
C2H5OH: [2(4)+6(1)+1(-2)]/2 = 12/2=6
High degree of reduction means low degree of oxidation CH4 > C2H5OH > C6H12O6

wS-4a=c B+fj p-------- (1)

a=1/4 (w S-c B- f j p)
From (1),
1=

fj p
4a
c
+ B+
w
w S w S
S

4a
= electron tranferred to O 2
w
S
c B
= electron tranferred to biomass
w S

fj p
w S

= electron tranferred to product

Maximum value of stoichiometric coefficient c (all electrons are used for biomass synthesis) is

c B
w S

& Cmax is given by

c max

Maximum value of stoichiometric coefficient f (no biomass forms) is

f max =

w
j

Q1. Production of single cell protein from hexadecane is given by the following reaction:

C16H34 + aO2 cCH1.66O0.27 N 0.2 + dCO2 + eH 2O

CH1.66O0.27N0.2 represents biomass , If RQ = 0.43, determine the stoichiometric coefficients.
Solution: C balance: 16 = c+d -----(1)
H balance: 34+ 3b = 1.66 c + 2e -----(2)
2a = 0.27c+2d+e ----------(3)
b= 0.2 c ------------(4)
RQ = d/a ------------(5)
Solving (1)-(5),
a=12.48
b= 2.13
c= 10.64
d=5.37
e=11.36

Q2. The respiration of glucose is given by:

C6 H12O 6 + 6O 2 6CO 2 + 6H 2O

Candida utilis cells convert glucose to CO2 and H2O during growth. The cell composition is
CH1.84O0.55N0.2 plus 5% ash. Yield of biomass from substrate is 0.5 g/g. NH3 is used as nitrogen
source.
(a) Find oxygen demand with growth and without growth.
(b) if ethanol is used as substrate to produce
(c) Cells of same composition as above, compare maximum possible biomass yields from
ethanol and glucose.
(d) Solution: (a) The cell composition is CH1.84O0.55N0.2 plus 5% ash. This means 95% of
total weight = 25.44
(e) Mwt. of biomass = 25.44/0.95= 26.78
(f) B = CH1.84O0.55N0.2 = (4x1)+ (1x1.84)-(2x0.55) (3x0.2) =4.14
(g) s1 = C6H12O6= 4, s 2= C2H5OH = 6
(h) Yxs = 0.5 g/g =(0.5/26.78)/(1/180)
(i) = 3.36 gmol/gmol
(j) a=1/4 (w S-c B- f j p), fjp =0 as no products are produced
(k) =1/4[6(4)-3.36(4.14)] =2.52
(l) Oxygen demand with growth/without growth = (2.52/6)x100 = 42%
(m) (b)
c max

C6H12O6 as substrate:
Cmax(C6H12O6) = 6(4)/4.14 = 5.8 gmol/gmol
YXS max = (5.8x 26.78)/(1x180)= 0.86 g/g
C2H5OH as substrate:
Cmax(C2H5OH) = 2(6)/4.14 = 2.9 gmol/gmol

YXS max = (2.9x 26.78)/(1x46)= 1.69 g/g

(Yxs)based on C2H5OH/ (Yxs)based on C6H12O6 = 1.69/2.9 = 1.96 2
Q3. Assume that the experimental measurements for a certain organism have shown that the cells
can convert 2/3 of substrate carbon to biomass. (a) Calculate the stoichiometric coefficients of
following reactions:
C16 H 34 + aO 2 + bNH3 cCH1.66O 0.27 N1.2 + dH 2O + eCO 2
(b) Calculate the yield coefficients YX/S, YX/O2 for both reactions
C6 H12O 6 + aO 2 + bNH 3 cCH1.66O 0.27 N1.2 + dH 2O + eCO 2
For C16H34 , C present = 16x12=192 g
C converted to biomass = 2/3(192) = 128 g
C balance: 128 = c(4.4)(12), c= 2.42
C converted to CO2= 192-128 = 128 g
64 = e(12), e=5.33
N balance: 14 b = c(0.86) (14), b = 2.085
H balance 34(1)+3b =7.3c+2d, d=11.29
O balance: 2a(16)=1.2c(16)+2e(16)+d(16)
a=12.427
For C6H12O6 : C present = 72 g
C converted to biomass = 2/3(72) = 48 g
48=4.4c(12), c=0.909
C converted to CO2 = 72-48=24g
24=12 e, e=2
N balance= 14b = 0.86c(14), c=0.782
H balance: 12+3b=7.3c+2d
d = 3.854
The oxygen balance yields

6 (16) + 2(16)a = 1.2(16)c + 2(16)e + 16d

a = 1.473
b, for hexadecane,

(YX/S)hexadecane > (YX/S)glucose

(YX/O2)glucose > (YX/S)hexadecane
Classification of microbial products

1) Growth associated: specific rate of product formation specific growth rate

ex: production of a constitutive enzyme
2) Non growth associated product: takes place in stationary phase and when growth rate is
zero. qP = = constant
Ex: secondary metabolite production of penicillin
3) Mixed growth associated product: takes place in slow growth and stationary phases. qP =
g +
ex: lactic acid fermentation, and xanthan gum production
Luedeking-Piret model
In the eq: qP = g + ,
If =0, qP = , product is only nongrowth associated
If =0, qP = g product is only growth associated

A strain of mold was grown in a batch culture on glucose and the following data were obtained.

Time (h)

Cell concentration (g/l)

Glucose concentration (g/l)

1.25

100

2.45

97

16

5.1

90.4

23

10.5

76.9

30

22

48.1

34

33

20.6

36

37.5

9.38

40

41

0.63

a. Calculate the maximum net specific growth rate

b. Calculate the apparent growth yield
c. C.
What maximum cell concentration could one except if 50 g of glucose were used
with the same size inoculum?
d.

Solution A plot of ln X versus t yields a slope of 0.1 h.

MODELS WITH GROWTH INHIBITORS

Substrate Inhibition
The microbial growth rate at higher substrate concentrations is said to be inhibited
by the substrate
Competitive Substrate inhibition :

Non competitive substrate inhibition:

Or if

Ks, then:

Product Inhibition
Competitive product inhibition:

Noncompetitive product inhibition :

Example of Non-competitive
competitive product inhibition
Ethanol fermentation from glucose

Where, Pm is the product concentration at which growth stops, or

where Kp is the product inhibition constant.

Inhibition by toxic Compounds

Competitive inhibition:

Noncompetitive inhibition :

Uncompetitive inhibition :

The net specific rate expression in the presence of death has the following form:

Where

is the death rate constant (h-1)

HEAT GENERATION BY MICROBIAL GROWTH

About 40% to 50% energy that is stored in a carbon and energy source is converted to biological
energy (ATP) during aerobic metabolism and the rest of the energy is released as heat The
actively growing cells, requires less maintenance, where the heat evolution is directly related to
growth.
The heat generate during microbial growth is being calculated using the heat of the combustion
of the substrate and
nd of cellular material. A schematic of an enthalpy balance for microbial
utilization of substrate is presented below as figure. The heat of combustion the substrate is equal
to the sum of the metabolic heat and the heat of combustion of the cellular material.
mater

The above equation is rearranged as to yield

The total rate of heat evolution in a batch fermentation is

Where VL is the liquid volume (I) and X is the cell concentration (g/l).Since the oxygen is
the final electron acceptor

Here QGR is in units of kcal/h, while QO2 is in millimoles of Q2/h

Metabolic heat released during fermentation can be removed by circulating cooling water
through a cooling coil or cooling jacket in the fermenter. Often, temperature control (adequate
heat removal) is an important limitation on reactor design. The ability to estimate heat-removal
requirements is essential for proper reactor design.

Reference Books
1) Shuler and Kargi (2004). Bioprocess Engineering:Basic Concepts, 2nd ed. Prentice Hall.
2) Doran P.M. (2005). Bioprocess Engineering Principles, 1st ed. Academic Press