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Lecture 6: Product Formation

Stoichiometry
Product Formation Stoichiometry
Product formation are classified into four classes

1. Main product appear as a result of primary energy


metabolism. Example : ethanol production during
anaerobic growth of yeast
2. Main product arises indirectly from energy metabolism.
Example: citric acid formation during aerobic mold
formation
3. Secondary metabolite. Example: penicillin production
in aerated mold culture
4. Biotransformation. The product is obtained from
substrate through one or more reactions catalyzed by
enzyme in the cells. Example: steroid hydroxylation
1 mole of carb
1 mole of cellular
material

CH m O n a O 2 b NH 3
c CH O N d . H 2 O e CO 2

Elemental
Balances
Degree of Reduction
Case 2: the simple stoichiometry does not apply. The
product formation is not proportional to substrate
utilization or cell mall increase; provides no insight into
the energetics of rxn

In complex reactions formation of extracellular pdts


Additional stoich coeff is needed

Degree of reduction = used for proton-electron balances


in bioreactions
Class 3 and 4: The stoichiometric depend on particular
substrate or products involved. Product formation is
completely uncoupled with cell growth.

Example, anaerobic fermentation of glucose to ethanol.


The best possible case is utilization of all substrate for
product formation alone. Assuming cell growth is
negligible
1 2
CH 2 O CO2 CH3O 0.5
3 3
1 2
CH 2 O CO2 CH3O 0.5
3 3
2/3 of the substrate carbon are used for product
formation.
Yield factor can be write as

(Yp s ) max 2 / 36 / 2 2
The upper bound for product yield can determine by
assuming that no cells or other products accumulate. For
example, for penicillin synthesis
1.5 glucose H 2SO 4 2NH3 phenylacet ate
penicillin G CO2 8H 2O
The maximum yield of penicillin is 1.2 g penicillin/(g
glucose). But the actual value is much more lower (0.1
g/g), indicating useful remaining potential for organism
and process improvement.
Yield much lower than upper bound value indicate that
significant substrate utilization supports growth,
maintenance or synthesis of other products
Metabolic Energy Stoichiometry: Heat
Generation and Yield Factor Estimation
Cellular heat production is the result of energy and
growth metabolism. It is proportional relationship
between heat generated and energy substrate utilized.

If Ys is grams of cell mass produced per gram substrate


consumed and DHs and DHc are heat of combustion of
substrate and of cell mass material, we can write yield
factor
Ys (g cell/g substrate)
YD (g cell/kcal)
DH Ys DH c kcal/g substrate
This arises from two energy balance pathways
the heat generation DHs per gram of substrate
completely oxidized minus YsDHc, the heat obtained by
combustion of cell grown from same amount of
substrate, will approximate the heat generation per gram
of substrate consumed in the fermentation which
produce the cell, H2O and CO2.
Example: The heat of combustion of pseudomonas
fluorescens growing in glucose medium. First a reaction
of cell combustion is written using measured cell
composition, assuming combustion products are H2O,
CO2 and N2.
CH1.66 N 0.22O0.27 1.28O2 CO2 0.10 N 2 0.83 H 2O
Cells
Assuming a heat of combustion of 104 kcal/mole O2, the
heat released by combustion of bacteria is

1.28 mol O2 104 kcal/mol O2 6.41 kcal


12 1.661 0.2214 0.2716g g
If cell dry matter contain 10% ash, then the heat of
combustion is

6.41 kcal
DH c 0.90
kcal

5.8
g g dry cell mass
A yeast (CH1.66N0.13O0.40) is growing aerobically on arabinose
(C5H10O5) and ammonium hydroxide (NH4OH) with a
respiratory quotient of 1.4.

1. Write the stoichiometric equation


2. Determine the yield coefficient, YX/S, for this system
in grams biomass/gram substrate
3. What is the maximum biomass concentration that
can be achieved in batch growth for this system if
the initial substrate concentration is 10 g/L and the
initial biomass concentration is 0.5 g/L
aC5H10O5 + bO2 + cNH4OH
CH1.66N0.13O0.40 + dCO2 + eH2O

Equations for coefficients:


C atom balance: 5a = 1 + d (1)
H atom balance: 10a + 5c = 1.66 + 2e (2)
O atom balance: 5a + 2b + c = 0.40 + 2d + e (3)
N atom balance: c = 0.13 (4)
Respiratory quotient: RQ = 1.4 = d/b (5)
2(3) - (2)
b = 0.2938
(5)
d = 0.4113
(1)
a = 0.2823
(3)
e = 0.9065
Stoichiometry of the equation

0.2823C5H10O5 + 0.2938O2 + 0.13NH4OH


CH1.66N0.13O0.40 + 0.4113CO2 + 0.9065H2O