Tutorial 1-H83BCE

Dr MR Kosseva

Stoichiometry of Growth

Elemental Balances for Microbial Reactions

Cell growth obeys the law of conservation of matter. For

many microorganisms, the energy and carbon requirements for
growth and product formation can be met by the same organic
compound.

Our starting point is a macroscopic view of the uptake of

nutrients (substrates for cell growth) and subsequent production
of new cells and products.

Some organisms are able to grow on a simple medium generally

containing a sugar source, inorganic nitrogen and a mix of salts.
The growth limiting substrate will generally be the first one to
be exhausted when the organism is grown batch-wise. We shall
consider a single growth limiting substrate to limit both
kinetically and stoichiometrically.

The cell formula is based on one gram-atom of carbon and

contains only the major elements. We shall write a formula for a
cell as CHaObNc, and ignore for the moment the other minor
elements in the cell (phosphorous, sulphur and ash).

When the cells grow on ammonia as the only source of nitrogen,

and carbohydrate as sole carbon and energy source, a general
form of the reaction resulting in aerobic cell growth and
formation of products can be written:

1
Consider aerobic cell growth:
Carbohydrate source of carbon and energy
Ammonia - source of nitrogen
Ignore phosphorous, sulphur and ash
CHlOm + NH3 + O2 CHaObNc +CHpOqNr +
H2O + CO2 (1)

The cell formula is based on one gram-atom of carbon

and contains only the major elements.

CHaObNc elemental composition of the cell

CHlOm elemental composition of the carbon source

CHpOqNr - elemental composition of extracellular product

The stoichiometric coefficient of the cells has been taken as
unity for simplicity.

Atomic balance equations can be written for C, H, O and N:

C: =1++

H: l + 3 = a + p + 2

O: m +2 = b + q + + 2

N: = c + r

2
A solution for 6 unknown stoichiometric coefficients can not be
obtained from these 4 atomic balance equations. Additional
empirically-based relationship is provided in aerobic growth by
respiratory quotient. This is defined as the rate of CO2
formation divided by the rate of oxygen consumption.

RQ = rate of CO2 production / rate of O2 consumption

(mM/liter-hr)

RQ = / (2)

Another relationship to determine unknown stoichiometric

coefficients can be obtained via electron balances.

The degree of reduction of an organic compound is defined

as the number of electrons available (per g atom Carbon) for
transfer to oxygen on combustion of a compound to CO2,
H2O, and N2.

The number of equivalents of available electrons is taken as

4 for carbon,
1 for hydrogen,
-2 for oxygen,
3 for nitrogen.

The degree of reduction of biomass, substrate and product is

b, s, p.

Substrate : s = 4 + l 2m CHlOm

Cells: b = 4+ a 2b 3c CHaObNc

Product: p = 4 + p - 2q 3r CHpOqNr
Note: CO2, H2O and NH3 have degree of reduction = 0

3
From reaction (1)

s 4 = b + p re-arranging,

1 = 4/ s + b/ s + p/ s (3)
oxygen cells product

Biomass Yield = Yxs = g Cells produced / g S consumed

Product Yield from S = Yps = g Product formed/ g S consumed

where S is the substrate concentration.

Hence, Oxygen requirements () can be calculated from (3).

References:
1. Blanch H.W. and Clark D.S. (1996) Biochemical
Engineering, Marcel Dekker, Inc., New York.
2. Doran P. M. (2013) Bioprocess Engineering principles.
Second Edition. Elsevier.
3. Roels, J.A. (1983) Energetics and Kinetics in
Biotechnology, Elsevier, Amsterdam.