Karan Damania BT19003

Test 4
Assignment 
a)Types of Fermentation
b) Process Parameters

Answer all the questions or any 5

1. Write a distinction between a. Anaerobic and Aerobic fermentation

b. Submerged and Surface fermentation
c. Batch fermentation and continuous fermentation

Answer:

Aerobic fermentation Anaerobic fermentation

 Set of chemical reactions involved in the  Chemical breakdown of organic substrates

production of energy by completely oxidizing into ethanol or lactic acid by microorganisms
food. in the presence of oxygen.
Fermentation  occurs in both cytoplasm and Fermentation occurs in the cytoplasm.
mitochondria.

Aerobic Fermentation: Occurs in higher Anaerobic Fermentation: Occurs in yeast,

animals and plants. parasites, and bacteria.

Aerobic Fermentation: Uses molecular Anaerobic Fermentation: Does not use

oxygen as the final electron acceptor in the
electron transport chain.
Produces six water molecules per glucose
molecule as its product.
ATP is a yield during the NAD+ regeneration .
The number of ATP produced is 36 in the
process
oxygen during the process.
Does not produce water as its product
 ATP is not a yield during the
NAD+ regeneration.
The number of ATP produced is 2 in the
process

Submerged Fermentation Surface Fermentation

Fermentation may be carried out as batch Fermentation may be carried out as a

or continuous. batch.
Aeration and agitation is of the system is Aeration is usually carried out by passing
possible by use of sparger and impeller. sterile air and no agitation.

Product yield are usually high as compared
to input cost.
If a batch gets contaminated there is a loss
of entire batch.
Entire fermentation media is utilized by
microorganism for growth and product
formation.
Inoculum is usually in liquid form.
Controlling parameters like temperature
and ph is easy
Product yield is comparatively less.
if a tray gets contaminated then there is
loss of only one tray but not the whole
batch.
There is wastage of fermentation media.
Inoculum is usually sprayed on surface of
the medium
Controlling parameters like temperature
and ph is difficult

 Batch fermentation continuous fermentation

Suitable for the production of secondary Suitable for the primary metabolites whose
metabolties whose production is not production is associated with growth of the
associated with growth of the microbes. organism.
More common method for large scale Less commonly used for large production.
production of cell biomass and products.
Nutrients in the fermenter are utilized in Nutrients in the fermenter are utilized in
relatively slow rate and chances of relatively fast rate and chances of
contamination less. contamination more
Nutrients are added only once and not Nutrients are added many times ( in the
added in between the fermentation beginning and in between) the
fermentation process.
Less control over the growth of the More comtrol on the growth and
microbes and the production of desired production.
products.
Setup is not changed from outside once the Setup is changed from outside during the
fermentation is started fermentation process.
The process is stopped once the product is The process is not for the collection of the
formed. products, but it is continuously taken out
from the fermenter.
 2. What is a microbial growth curve, explain in detail

When a broth culture is inoculated with a small bacterial inoculum, the population size of the
bacteria increases showing a classical pattern. When plotted on a graph, a distinct curve is obtained
referred to as the bacterial growth curve.

Method of Obtaining Bacterial Growth Curve:

 A population growth curve for any particular species of bacterium may be determined by
growing a pure culture of the organism in a liquid medium at a constant temperature.
 Samples of the culture are collected at fixed intervals (e.g., every 30 minutes), and the
number of viable organisms in each sample is determined.
 The data are then plotted on logarithmic graph paper.
 The logarithm of the number of bacteria per milliliter of medium is plotted against time.

The bacterial growth curve shows the following four distinct phases:

1. Lag phase:
  After a liquid culture broth is inoculated, the multiplication of bacteria does not start
immediately. It takes some time to multiply.
 The time between inoculation and beginning of multiplication is known as lag phase.
 In this phase, the inoculated bacteria become acclimatized to the environment, switch on
various enzymes, and adjust to the environmental temperature and atmospheric conditions.
 During this phase, there is an increase in size of bacteria but no appreciable increase in
number of bacterial cells. The cells are active metabolically.
 The duration of the lag phase varies with the bacterial species, nature of culture medium,
incubation temperature, etc.
 It may vary from 1 hour to several days.

2. Log phase:

 This phase is characterized by rapid exponential cell growth (i.e., 1 to 2 to 4 to 8 and so on).
 The bacterial population doubles during every generation. They multiply at their maximum
rate.
 The bacterial cells are small and uniformly stained.
 The microbes are sensitive to adverse conditions, such as antibiotics and other
antimicrobial agents.
 Growth rate is the greatest during the log phase.
 The log phase is always brief, unless the rapidly dividing culture is maintained by constant
addition of nutrients and frequent removal of waste products.
 When plotted on logarithmic graph paper, the log phase appears as a steeply sloped straight
line.

3. Stationary phase:

 After log phase, the bacterial growth almost stops completely due to lack of essential
nutrients, lack of water oxygen, change in pH of the medium, etc. and accumulation of their
own toxic metabolic wastes.
 It is during this phase that the culture is at its greatest population density.
 However, Death rate of bacteria exceeds the rate of replication of bacteria.
 Endospores start forming during this stage.
 Bacteria become Gram variable and show irregular staining.
 Many bacteria start producing exotoxins.
4. Decline phase:

 During this phase, the bacterial population declines due to death of cells.
 The decline phase starts due to
(a) accumulation of toxic products and autolytic enzymes and
(b) exhaustion of nutrients.
 Involution forms are common in this stage. Some cells assume various shapes, becoming
long, filamentous rods or branching or globular forms that are difficult to identify.
 Some develop without a cell wall and are referred to as protoplasts, spheroplasts, or L-
phase variants (L-forms).
 When these involuted forms are inoculated into a fresh nutrient medium, they usually
revert to the original shape of the healthy bacteria.
 4. Write a note on anaerobic fermentation

Occurring in the absence of oxygen or not requiring oxygen to live. Anaerobic bacteria produce
energy from food molecules without the presence of oxygen.

Anaerobic fermentation occurs in the fermentation vessel once the oxygen is discharged and
replaced with N2, CO2, or another by-product of the fermentation process. Anaerobic fermentation
is usually a slower process. He showed that a microorganism, probably Clostridium butyricum, was
responsible for butyric acid fermentation. During World War I, industrial anaerobic fermentation
was further demonstrated by Perkins and Weizmann, who worked on acetonebutanol-ethanol
(ABE) fermentation with C. acetobutylicum.

Anaerobes may grow under the unfavorable conditions used to minimize contamination during
fermentations because they have unusual enzymes and catabolic pathways. Most anaerobic
fermentations require little energy to keep cells in suspension. Because less biomass is produced in
anaerobic fermentations, more carbon can be converted to the end product, and a higher product
yield is attained. Anaerobes can utilize a wide range of substrates, including agricultural waste
streams. This reduces the overall cost of the fermentation process. Anaerobic fermentation has
been applied to many important industrial fermentations, such as ethanol production by yeasts,
lactic acid preservation of foods, anaerobic digestion of organic matters in ruminant cultivation and
waste treatment. The most important industrial fermentation is the anaerobic production of
ethanol by S. cerevisiae and other yeasts.

However, mixed-culture processes in anaerobic fermentation are also difficult to study and model.
The microbial communities are usually unstable, varying with environmental changes and the
availability of nutrients. Obligate anaerobes need specialized media and apparatus. They are
deactivated by exposure to oxygen. Hence, special skills and meticulous methods are required for
the cultivation and manipulation of strictly anaerobic microorganisms. Compared to aerobic
organisms, there is little known about methods for genetic manipulation and to express desired
genes or biosynthetic pathways.
 5. Explain the importance of Submerged fermentation

Answer- In the submerged process, the substrate used for fermentation is always in liquid state
which contain the nutrients needed for growth.

The fermentor which contain the substrate is operated continuously and the product biomass is
continuously harvested from the fermenter by suing different techniques then the product is
filtered or centrifuged and then dried.

Submerged fermentation is a method of manufacturing biomolecules in which enzymes and

other reactive compounds are submerged in a liquid as alcohol, oil, or a nutrient broth.

Submerged fermentation utilizes free flowing liquid substrates, such as molasses and broths.
The bioactive compounds are secreted into the fermentation broth.

The substrates are utilized quite rapidly hence need to be constantly replaced supplemented
with nutrients.

This fermentation technique is best suited for microorganism such as bacteria that require high
moisture. An additional advantage of this technique is that purification of products is easier.

Submerged is primarily used in the extraction of secondary metabolites that need used in liquid
form.

9. Explain the use of biosensors to conduct a fermentation.

Answer- In fermentation process safety and product quality are crucial. Thus effective
monitoring od the fermentation process is imperative to develop, optimize, and maintain
biological reactors at maximum efficiency.

Biosensor can be utilized can be utilized to monitor this presence of products biomass enzymes,
antibody or by products of the process to indirectly measure the process conditions.

Biosensor precisely control the fermentation industry and produce results due to their simple
instrumentation, formidable selectively, low prices and easy automation.

Now a days, several kinds of commercial biosensor are accessible capable of detecting
biochemical parameters (glucose, lactose, lysine, ethanol etc) and are widely used in chna
occupying 90% of its market. since the launch of glucose biosensor commercially in 1975 the
fermentation industries have been benefitted.
Now the factories successfully use glucose biosensor to control production in the fermentation
process and utilize the bio enzymatic method to produce glucose. Biosensor are employed in
ion exchange retrival, where detection of change of biochemical composition is carried out.

Glutamic biosensor has been used to conduct experiments on ion exchange reterival of an
isoelectric liquor supernatural of glutamic. In past years have attracted a lot of attention in
online monitoring fermentation process due to its simplicity and quick response.