Fermentation Technology 1

Dr. Nermin Hassan Ibrahim

Ass. Prof. of Medical Microbiology
and Immunology
What is fermentation?

Pasteur’s definition: “life without air”, anaerobe redox

reactions in organisms

New definition: a form of metabolism in which the end

products could be further oxidized

For example: a yeast cell obtains 2 molecules of ATP per

molecule of glucose when it ferments it to ethanol
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What is fermentation techniques (1)?
Techniques for large-scale production of microbial products. It
must both provide an optimum environment for the microbial
synthesis of the desired product and be economically feasible on
a large scale.
They can be divided into surface (emersion) and submersion
techniques.
The latter may be run in batch, fed batch, continuous reactors
In the surface techniques, the microorganisms are cultivated on
the surface of a liquid or solid substrate. These techniques are
very complicated and rarely used in industry

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What is fermentation techniques (2)?

In the submersion processes, the microorganisms grow in a

liquid medium. Except in traditional beer and wine fermentation,
the medium is held in fermenters and stirred to obtain a
homogeneous distribution of cells and medium.
Most processes are aerobic, and for these the medium must be
vigorously aerated.
All important industrial processes (production of biomass and
protein, antibiotics, enzymes and sewage treatment) are carried
out by submersion processes.

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Some important fermentation products
Product Organism Use

Ethanol Saccharomyces Industrial solvents,

cerevisiae beverages
Glycerol Saccharomyces Production of
cerevisiae explosives
Lactic acid Lactobacillus Food and
bulgaricus pharmaceutical
Acetone and Clostridium Solvents
butanol acetobutylicum
-amylase Bacillus subtilis Starch hydrolysis

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Some important fermentation products

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Some important fermentation products

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Some important fermentation products

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Wine-making fermenter

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General Aspects of Fermentation
Processes
All cells break down complex organic compounds into
simpler molecules. Cells use some of the energy that
is released in this process to make ATP.

What is cellular respiration?

Cellular respiration- is a complex process in which cells

make ATP by breaking down organic compounds.

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 What is glycolysis?

Glycolysis- is the first biochemical pathway, which yields

small amounts of ATP.

Glycolysis consist of two pathways:

1. Glycolysis Oxygen present
Aerobic respiration ATP

2. Glycolysis Oxygen Absent

Fermentation (anaerobic).
C6H1206 → 2 CH3CH2OH + 2 CO2 + 2 ATP
C6H1206 + 6O2 → 6CO2 + 6H2O + 16-18 ATP

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What is anaerobic respiration?

Anaerobic respiration- makes ATP without the use of oxygen, instead

uses sugar.
 
What is aerobic respiration?

Aerobic respiration- is the process of making ATP with the use of

oxygen and produces much larger amount of ATP than anaerobic
respiration.

So Glycolysis can be a pathway in which one six-carbon molecule of

glucose is oxidized to produce two three- carbon molecules of Pyruvic
acid.

Aerobic respiration has two major stages: Krebs cycle, and the
electron transport chain. 13
 Glycolysis

Substrate-level phosphorylation

2 pyruvate
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Fermentation is an energy yielding process where
organisms converts organic molecules (such as
sugar) into energy, carbon dioxide or/and ethanol
depending on the respiration pathway.

Fermentation has been practiced for years and has

resulted in synthesis of different kinds of food such as
bread, wine and beer.

There is two types of fermentation:

1) Homolactic fermentation.
2) Ethanol Fermentation.

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  Homolactic fermentation:

• Homolactic Fermentation breaks down the pyruvate into Lactate.

• It occurs in the muscles of animals when they need energy faster than the

blood can supply oxygen.

• It also occurs in some kinds of bacteria (such as lactobacilli) and some fungi.

• It is this type of bacteria that converts lactose into lactic acid in yogurt, giving

it its sour taste.

• These lactic acid bacteria can be classed as homofermentative, where the end

product is mostly lactate, or heterofermentative, where some lactate is

further metabolized and results in carbon dioxide, acetate or other metabolic

products
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Ethanol Fermentation :

• Ethanol fermentation (performed by yeast and some types of

bacteria) breaks the pyruvate down into ethanol and carbon dioxide.

• It is important in bread-making, brewing, and wine-making.

Usually only one of the products is desired; in bread-making, the

alcohol is baked out, and, in alcohol production, the carbon dioxide

is released into the atmosphere or used for carbonating the

beverage.

• When the ferment has a high concentration of pectin, minute

quantities of methanol can be produced.

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 Properties of an ideal Industrial micro-organisms

Often fungi or streptomycetes (bacteria)

Grow rapidly & produce product in relatively short period.

Organisms can be manipulated in large-scale cheap culture to

produce one or more products in high concentration.

Organisms are genetically stable with infrequent mutation

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Able to grow in inexpensive liquid culture media (waste

from other industries): eg

• Corn steep liquor from corn milling (rich in N &

growth factors)
• Whey from cheese production (rich in lactose &

minerals)
 Should not be pathogenic (humans, animals, plants)

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Fermentation medium
Medium  nutritional, hormonal, and fulfilling

requirement of cells
 In most cases, the medium is independent of the

bioreactor design and process parameters

 Even small modifications in the medium could

change cell line stability, product quality, yield,

operational parameters, and downstream processing.
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THINK-PAIR AND SHARE
Summarize in 2
sentences
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)
 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
 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.
 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
 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
 Metabolites
Metabolites are the intermediates and products of metabolism.
The term metabolite is usually restricted to small molecules.
 A primary metabolite is directly involved in normal growth,
development, and reproduction.
 A secondary metabolite is not directly involved in those
processes, but usually has an important ecological function.
Examples include antibiotics and pigments.
• Primary metabolite :
formed during growth phase
• Secondary metabolite:
formed near the end of growth phase frequently during near
or in stationary phase
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 Secondary metabolites
produced from substrates provided by primary metabolism

 not essential for growth and reproduction.

formation is extremely dependent on growth conditions.

often produced as a group of closely related products

e.g. a particular streptomycete strain is known that can produce > 30

related but different anthracycline antibiotics

 possible to induce dramatic overproduction of secondary metabolites

(different from primary metabolites which usually cannot be

significantly overproduced).

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Most secondary metabolites are complex organic

molecules requiring a large number of specific

enzymatic reactions for synthesis

e.g. tetracycline synthesis (72 separate enzymatic

steps).

e.g. erythromycin synthesis ( 25 steps)

No one organism produces all of these secondary

metabolites & there are many steps between the amino

acid & the antibiotic
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Thank you
 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
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

 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
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
 Pharmacy college Pharmaceutics/ Microbiology Department

Dr. Nermin Hassan Ibrahim

1432 -1431 ‫جدول الفصل الدراسى االول‬

Day 8-9 9-10 10-11 11-12 12-1 1-2 2-3 3-4

Saturday Office Biotech. Office Academic
hour 424 hour advisor

Sunday Academi Research activity 226 MIC I

c advisor

Monday Office Biotech. Academic

hour 424 advisor

Tuesday Biotech. 226 MIC I 226 MIC I practical

424
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Thursday Office
hour
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advisor Research activity