Introduction to

Fermentation
Aspergillus niger and Lactobacillus
 Introduction to Fermentation
• Aspergillus niger and Lactobacillus Delbruckii are the
microbes used to commercially produce citric acid and
lactic acid, respectively. The production takes place in a
batch fermenter. This tutorial will introduce you to the
following areas regarding batch fermentation
– Microbial Growth Kinetics
– Media for Industrial Fermentations
– Sterilization
– The Development of Inocula for Industrial Fermentations
– Design of a Fermenter
– Instrumentation and Control
– Aeration and Agitation
 Microbial Growth Kinetics
• 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

(Growth curve is from Shuler p.

161)
 Microbial Growth Kinetics
• 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.
 Microbial Growth Kinetics
• 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
 Microbial Growth Kinetics
• Exponential Phase (Continued)
– At some point the cell growth rate will level
off and become constant
– The most likely cause of this leveling off is
substrate limited inhibition
• Substrate limited inhibition means that the
microbes do not have enough nutrients in the
medium to continue multiplying.
 Microbial Growth Kinetics
• 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
 Microbial Growth Kinetics
• Death phase
– The fourth phase in the fermentation process
– The number of cells dying is greater than the
number of cells multiplying
• The cause of the death phase is usually that the
cells have consumed most of the nutrients in the
medium and there is not enough left for
sustainability
Media for Industrial Fermentations
• The media is the feed solution
– It must contain the essential nutrients needed for the
microbe to grow
• Factors of consideration when choosing media
-Quality consistence and availability
-Ensure there are no problems with Media Prep or
other aspects of production process

Ex. Cane molasses, beet molasses, cereal grains

 Sterilization
• Sterilizing the feed solution is essential
because the media cannot contain foreign
microbes because this could severely
hinder the growth of the production
microbe
– Most popular method is heat sterilization of
the feed solution
 The Development of Inocula for
Industrial Fermentations
• The inoculum is the starter culture that is
injected into the fermenter
– It must be of sufficient size for optimal growth
kinetics
• Since the production fermenter in industrial
fermentations is so large, the inoculum volume
has to be quite large
- A seed fermenter is usually required to produce the
inoculum volume
-The seed fermenter’s purpose is not to produce
product but to prepare inoculum
 Design of a Fermenter
• Factors to consider when
designing a fermenter
– Aseptic and regulatory
capability, long-term
reliability
– Adequate aeration and
agitation
– Low power consumption
– Temperature and pH
controls
– Sampling facilities

(14 L fermenter shown is a

copyright of New
Brunswick Scientific)
 Instrumentation and Control
• The success of a fermentation process is
highly dependent on environmental
factors
– The fermenter needs to be able to control
such factors as temperature, pH, and
dissolved oxygen levels
 Aeration and Agitation
• Most industrial fermentations are aerobic
processes meaning that the production
microbe requires oxygen to grow
– The oxygen demand is met by sparging air
through the fermentation vessel and using an
agitator increase the amount of dissolved
oxygen
 References
• Stanbury, P.F., A. Whitaker, and S. J. Hall,
Principles of Fermentation Technology, 2nd
ed., Butterworth Heinemann, Oxford,
2000.
• Shuler, M. L. and F. Kargi. Bioprocess
Engineering Basic Concepts, 2nd ed.,
Prentice Hall, Upper Saddle River, NJ,
2002.