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STOICHIOMETRY OF

MICROBIAL
GROWTH AND PRODUCT
FORMATION
Microbial growth stoichiometry involves
the conversion of substrates into
products and cellular materials and
helps to determine the true yield
coefficient based on substrate for
biomass (Yx/s) and for product (Yp/s).
FIRST strategy for a rational design of a medium is
according to the elementary composition of a specific
micro-organism and of the main product that is
produced.
SECOND is to design according to the stoichiometry
of growth and product formation which is represented
by a chemical equation.
Yield Coefficients
- are defined based on the amount of consumption
of another material
- used to convert between cell growth rate and
substrate utilization

Yx / S = cells produced / substrate consumed


= (molwt. cells) / (molwt. substrate)

Yp / S = product produced / subtsrate consumed


= (molwt. product) / (molwt. substrate)
STOICHIOMETRIC CALCULATIONS

ELEMENTAL DEGREE OF
BALANCES REDUCTION
Simple elemental balances on C, H, O and
N yield the following equations:

Basis of 1 mole of
carbohydrate
Typical
When cellular
little info is composition:
available about cell
composition, use the approximated cell
CH 1.8O0.5N0.2
composition.

Different types of organisms have different cellular composition.


SAMPLE PROBLEM
Assume that experimental measurements for a certain
organism have shown that cells can convert 2/3(wt/wt) of the
substrate carbon (alkane or glucose) to biomass.

Hexadecane
C16H34 + a O2 + b NH3 → c C4.4H7.3O1.2N0.86 + d H2O + eCO2
Glucose
C6H12 O6 + a O2 + b NH3 → c C4.4H7.3O1.2N0.86 + d H2O + eCO2

a. Calculate the stoichiometric coefficients for the following


biological reactions.
b. Calculate the yield coefficients Yx/s (gdw cell/ g
substrate) for both reactions.
Hexadecane
C16H34 + a O2 + b NH3 → c C4.4H7.3O1.2N0.86 + d H2O + eCO2

Glucose
C6H12 O6 + a O2 + b NH3 → c C4.4H7.3O1.2N0.86 + d H2O + eCO2
Hexadecane
C in 1 mole of substrate = 192g Nitrogen balance:
C converted to biomass = 192(2/3) 14b = c (0.86) (14)
= 128g b = (2.42) (0.86)
Then, 128 = c (4.4) (12); c = 2.42 b = 2.085
C converted to CO2 = 192 -128
= 64g Oxygen balance:
64 = e (12) 2a (16) = 1.2c (16) + 2e (16) + d
e = 5.33 (16)
Hydrogen balance: a = 12. 427
34 (1) + 3b = 7.3c + 2d
34 + 3 (2.085) = 7.3 (2.42) +2d b. For hexadecane
d = 12.43 Y x/s = 2.42 (91.34) / 226
Y x/s = 0.98 gdw cells / g substrate
Glucose
C in 1 mole of substrate = 72g Hydrogen balance:
C converted to biomass = 72 (2/3) 12 + 3b = 7.3c + 2d
= 48g d= 3.854
Oxygen balance:
Then, 48 = 4.4c (12); c = 0.909 6 (16) + 2(16)a = 1.2 (16) c + 2 (16) e
C converted to CO2 = 72 – 48 + 16d
= 24g a = 1.473
24 = 12e
e=2 b. For glucose
Nitrogen balance: Y x/s = 0.909(91.34)/ 180
14b = 0.86c (14); b = 0.86 (0.909) Y x/s = 0.461 gdw cells/ g substrate
b = 0.782
Unfortunately, the elemental
balances often do not provide
enough information to completely
solve for the stoichiometric
coefficients.
Respiratory Quotient (RQ)

RQ = YCO2/O2
- The mole of CO2 produced per mole of O2 consumed.
- Provides information on the metabolic state of the cell.
- A high RQ means that much CO2 is produced and
hence the metabolism is operating at high efficiency.
Simple elemental balances on C, H, O and
N yield the following equations:

RQ = e/a
five equations for five
unknowns a, b, c, d and
e.
SAMPLE PROBLEM
A yeast (CH1.66 O0.40N0.13) is growing on arabinose
(C5H10 O5) and ammonium hydroxide (NH4OH)
with a respiratory quotient of 1.4.

1. Write the stoichiometric equation for 1mol of yeast.

a C5H10 O5 + bO2 + cNH4OH→ CH1.66O0.40N0.13 + d CO2


+ eH2O
2. Determine the yield coefficient Yx/S.

Equation for coefficients:


a = 0.2823
C balance: 5a = 1 + d
b = 0.2938
H balance: 10a + 5c = 1.66 + 2e c = 0.13
O balance: 5a + 2b + c = 0.40 + 2d +e d = 0.4113
N balance: c = 0.13 e = 0.9065
RQ = d/b = 1.4

Yx/S = (21.88 molwt yeast )/ (0.2823 x150 molwt arabinose)


= 0.5167 cells/ substrate
ELEMENTAL BALANCES
PROVIDE NO INSIGHT INTO
THE ENERGETICS OF A
REACTION.

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