Microbial Growth Phases

(Fermentation)
Lag Phase
Exponential Phase
Stationary Phase
Death Phase

Microbial Growth Phases

(Fermentation)

Lag Phase
The first phase of growth in a batch culture
A period of adaptation of the cells to their new

environment
Little increase in cell density
May not last long in some fermentations

Exponential Phase
Also known as the logarithmic growth phase
Cells are adjusted to their environment and are

dividing continuously
results in an exponential increase in the

number of cells
Cell growth rate is often substrate limited

Stationary Phase
When the number of cells dividing and dying is

the same
Depletion of one or more essential growth

nutrients
Accumulation of toxic by-products
Stress associated with the induction of a

recombinant gene

Stationary Phase
Primary growth associated production stops
Secondary, non-growth associated, production may

continue
This where the product is produced ($$$)
The stationary phase can also be induced chemically

- Sodium butyrate is an example of such a chemical

Death Phase
Also known as the decline phase
The rate of cells dying is greater than the rate

of cells dividing

Establishing a Growth Curve

Counting the Cells
Viable (living) Cell Count
Optical Density

Viable (living) Cell Count

represents the cells are actually living (more

accurate)
Counts made using a hemocytometer /trypan

blue stain under a microscope

Viable (living) Cell Count

or plating a sample

from the culture and

counting colonies

Optical Density (turbidity,

absorbance)
Counts made by taking an optical

measurement using a spectrophotometer

Measures living and dead cells (less accurate)
Different cell types are measured at different
wavelengths (i.e. E. coli @ 600nm)
The measurements are compared to reference
readings made previously

Spectrophotometers
New Model

Old Model

UV Vis

Light

Useful for trace analysis

Utilizes the Lambert-Beer Law:
logarithmic relationship of the

absorption of light in relation to the

concentration of the material it is
travelling through

Light spectrophotometers can be

dialed in to a desired wavelength for

a sample (400-700 nm)
Light of this wavelength then passes
through the sample
The intensity of the light that passes
through the sample is then measured
by a sensor or detector at the other
end

This intensity is translated in a

number by the spectrophotometer

This number can be read as

transmittance or absorbance
We like to use absorbance as it gives

a linear curve (straight line)

 The instrument should be

zeroed or blanked with a

cuvette containing only a clean
sample of the substance
This will serve as a baseline to

which other standards or samples

will be compared