STOICHIOMETRY OF MICROBIAL GROWTH

Objectives: Lecture 3
• Biosynthetic processes (anabolic)
• Case studies - cholesterol
• Stoichiometry and modeling cellular
requirements
• "You are what you eat"
 Stoichiometry
• Provides information on fundamental
constraints
– Substrate conversion to product
– Cell mass from substrate
 Yields and yield coefficients
• Mass based = “kg” of this from “kg” of that
• Y (output / input)
• Y x/s
• Y p/s
• Y ATP/O2

• Ymx/s maximal yield of cell mass from

substrate
 Yield
• Overall
• Instantaneous YIELD

• Ratio of rates
• Ratio of yields

• Theoretical = Y
• Observed = Y’
Cell metabolism

Y’ lactate / glucose = ranges from 2 to 0 based on environment

The basic reaction is:

Glucose + 2 Pi + 2 ADP → 2 Lactate + 2 ATP + 2 H2O

•Yield of cell mass from substrate
Y x/s

Bacterial
dry cell weight Slope = dX/dS
[mg/L] 7 (mg/L) / (g/L)

Glucose [g/L]
 Aerobic
Yx/s=58 mg/mol

Bacterial
dry cell weight
[g/L]
Anaerobic
Yx/s=22 mg/mol

Glucose [mM]
 Cell composition
Dry weight vs. wet weight
70% of the composition is water

CHxOyNz
Dry weight consists of:
Element E. coli Yeast

C 50% 50%
O 20% 34%
N 14% 8%
H 8% 6%
P 3% 1%
S 1% <1%
K 1% <1%
Na 1% <1%
Others <1% <1%
• In a very simplistic interpretation of
metabolism, the following applies:

– Cells + medium + O2 (sometimes) → more cells +

product + CO2 + H2O

• Medium contains sugars, amino acids,

cofactors and the elements in the previous
table.
1 mole of
1 mole of
carb
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
 Stoichiometric calculations
• Based on 1 mole of C in the input

• CHmOn + a O2 + b NH3 →

• c CHON + dH2O + eCO2

• This is normalized to 1 mole of C.

Could also be normalized to 1 mole of the C source compound

• Perform elemental balances to determine the

unknown values of the cofactors
 Example
• C6H12O6 + a O2 + b NH3 →

• c C4.4H7.3O1.2N0.86 + dH2O + eCO2

• 2/3 of the glucose C goes to biomass

• What are the stoichiometric coefficients,
and Yx/s, Yx/O2? MWglucose = 180
MWcell = 89.62
MWoxygen = 32
MWammonia = 17
 Generalized growth reaction
• C6H12O6 + a NH3 + b O2 →  CH1.8O0.5N0.2 +
  CHxOyNz + gCO2 + H2O

• Normalized to 1 mole of carbon source compound

• Where a, b, , , g, , x, y, z depend on the type of cell
involved.
• a, b, , , g, , are stoichiometric coefficients
• When little info is available about cell composition, use
an approximated cell composition of
CH1.8O0.5N0.2
• This yields a MW of a cell ~ 24.6
 Generalized growth reaction

C6H12O6 + a NH3 + b O2 →  CH1.8O0.5N0.2 +

 CHxOyNz + gCO2 + H2O

24.6
Yx s 
180
g of cells from g of glucose
 Lack of information
• Unfortunately, the elemental balances
often do not provide enough information
to completely solve for the stoichiometric
coefficients.
 Respiratory quotient
• RQ = YCO2/O2

• Molar basis
– Moles of CO2 produced from moles of O2

• 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
 Aerobic metabolism
• CHmOn + a O2 + b NH3 →

• c CHON + d CHxOyNz + eH2O + fCO2

• RQ = ?
 Degree of reduction
• Electron balance
g = # of available electrons / g of atomic C
g Or, this can be described as:
g = # of available electrons / # of C’s

• Provides another independent equation

 Degree of reduction
• C=4 • CO2 = +4 (C) + -2 (O) = 0
• H=1
• N = -3 • C6H12O6 = 6(4) + 12(1) + 6(-2) = 24
 g = 24 / 6 (# carbon atoms) = 4
• O = -2
• P=5 • C2H5OH = 2(4) + 6(1) + (-2) = 12
• S=6  g = 12 / 2 (# carbon atoms) = 6
 Example – yeast grown on glucose
• C6H12O6 + 0.48 NH3 + 3 O2 →

 0.48 CH1.8O0.5N0.2 + 3.12CO2 + 4.32H2O

• To grow yeast to 50 g/L in a 100,000 L reactor, determine:

• a) mass of glucose and ammonia required

• b) O2 required

• c) Yx/s and YX/O2

MWglucose = 180
MWcell = 24.6
MWoxygen = 32
MWammonia = 17
 HW #1 questions
1) What kind of cell would you use to produce
androstenedione? Your answer should
describe the attributes of such a cell (don't
just state, "a cell that produces andro"). An
answer longer than 4 sentences is too much.
2) Producing cholesterol is an energy intensive
process. How much energy (in terms of # of
ATP molecules) is consumed in producing
one cholesterol molecule from a source of
glucose?
3) Problem 7.1 - 7.7