Microbial Growth Kinetics describe how the microbe grows in the fermenter.
This information is important to determine optimal batch times. The growth of microbes in a fermenter can be broken down into four stages:
Lag Phase Exponential Phase Stationary Phase Death Phase
Apparatus
pH probe D-oxygen probe Mixer Temperature sensor Bioreactor YSI 2700 Biochemistry Analyzer
pH meter Sampling device
Using Biochemistry Analyzer and Spectrophotometer to measure and make calibration curves for sugar and yeast cell concentrations Reactant initial concentration
dextrose/or sucrose 25 g/L yeast 3 g/L volume reactant solution 2 L
Lag Phase
This is the first phase in the fermentation process The cells have just been injected into a new
environment and they need time to adjust accordingly Cell growth is minimal in this phase.
Exponential Phase
The second phase in the fermentation process The cells have adjusted to their environment and
rapid growth takes place Cell growth rate is highest in this phase
Stationary phase
This is the third phase in the fermentation process The cell growth rate has leveled off and become
constant The number of cells multiplying equals the number of cells dying
Death phase
The fourth phase in the fermentation process The number of cells dying is greater than the
number of cells multiplying
Growth Can be represented by:
Considering primary constituents:
Concept : Cell Yield
Experimental observation ; Cell mass is proportional to available substrate
Generalized substrate consumption and biomass growth with time.
Cell Growth - Kinetics
Growth rate to population size units are 1/t (i.e. h-1 1/h)
X = concentration of microorganisms at time t (g/l) t = time (h) = proportionality constant or specific growth rate, [h-1] dX/dt = microbial growth rate, [mass/volume time]
dX X dt
Doubling Time
Generation time or doubling time is the time required for the population to double. The calculation is: td = ln(2)/
Classified based on the relationship between product synthesis and energy generation in the cell Growth associated Non-growth associated Mixed-growth associated
Growth-associated
produced at the same time as cell growth
metabolic intermediates
pyruvate, citrate, acetate
Non-growth-associated
secondary metabolites
antibiotics
takes place during the stationary phase (=0)
Mixed - growth associated
takes place during growth and stationary phases
metabolic byproducts
lactate, ethanol
secondary metabolites
Growth-associated Mixed-growth-associated Non growth-associated
Rate per microbe, which depends on
Species Substrates
Environmental factors
Total numbers of microbes
Batch Growth Kinetics
Cell Yield is:
Yx / s
Change in Cell Mass Substrate Consumed
Yx / s
dX dS dP dS
Product yield is defined as:
YP / S
Growth Yield, Yield Coefficient
Growth yield = microorganisms produced per unit substrate utilized.
YX/S
X S
Other yield coefficients.
YX / O2
X O2
YP / S
P S
time (hr) 0 2 4 8 10 12 14 16 18
X (g/L) 0.34 0.36 0.52 1.67 3.01 5.44 9.52 10.46 10.54
S (g/L) 19.38 19.34 19.05 16.86 14.28 9.66 1.93 0.16 0.00
Find
Maximum growth rate
Yield on substrate doubling time specific growth rate at 12 hr
Example Problem Solution max
dX X dt
1 dX X dt
Integrate
ln(X) ln(Xo) = t
plot ln(X) vs. t
Slope (largest) = max
3 2.5 2 1.5
ln(X)
experimental data regression
1 0.5 0 -0.5 -1 -1.5 Time (hr) 0 5 10
slope=u max=0.292
15
20
td = ln 2/
Maximum doubling time will occur in exponential growth, when = max
Example Problem Solution
YX/S
X L 0.52 g cell g substrate S (0 19.38) g substrate L
(10.54 0.34) g cell
YP/S
Temperature effects on growth rate.
Classifications of microbes according to temperature optima.
Classification of microbes according to tolerance of pH extremes
Classification of microbes according to their oxygen responses.
a. b. c. d. e. Aerobic Anaerobic Facultative Microaerobic Aerotolerant
