About fermentation media

1. SRI PARAMAKALYANI COLLEGE
REACCREDITED WITH A+ GRADE WITH A CGPA OF 3.39 IN THE THIRD CYCLE OF NAAC
AFFILIATED TO MANOMANIUM SUNDARANAR UNIVERSITY, TIRUNELVELI.
ALWARKURICHI 627 412, TAMIL NADU, INDIA
POST GRADUATE & RESEARCH
DEPARTMENT OF MICROBIOLOGY
(Government Aided)
ACADEMIC YEAR 2022-2023
IV SEM CORE: INDUSTRIAL MICROBIOLOGY
UNIT- 1
FERMENTATION MEDIA & RAW MATERIALS IN
INDUSTRIAL FERMENTATION
Submitted by,
S.INDHUMATHI
Reg. No:20211232516110
II M.Sc. Microbiology
Assigned on: 27/01/2023
Taken on:02/03/2023
Submitted to,
Dr. S.VISWANATHAN Ph.D.
Assistant Professor and Head,
Department of Microbiology.
SPKC.

2. SYNOPSIS
Fermentation media & raw materials 2

3. Introduction
Fermentation, chemical process by which molecules such as glucose are broken
down anaerobically. The metabolic process that induces chemical changes in organic
substrates by the action of enzymes is known as fermentation. It uses microorganisms
such as bacteria, algae and fungi. The use of this process on a large scale to produce
pharmaceuticals, enzymes and proteins is known as industrial fermentation.
Fermentation (latin word) - "to leaven, cause to rise or ferment,"
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6. Characteristics of an ideal Culture medium
Satisfactory
growth for small
inoculum – even
for single cell.
Suitable pH
Rapid growth Nutrients
Cheap Easily producible
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Energy source Carbon source
Nitrogen source Salts
pH
Adequate
oxidation
Growth factors
Culture media - A growth medium or culture medium is a solid, liquid, or semi-
solid designed to support the growth of a population of microorganisms or cells
via the process of cell proliferation.

7. Components of an Culture Media
1. Water
It is essential for existence of living cells. They act as source of hydrogen
and oxygen.
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2. Peptone
Golden granular hygroscopic powder which are obtained from
meat, casein fibrin or soya bean flour.
Function: nitrogen source, carbon source, buffers
3. Meat Extract
It contains protein degradation products, carbohydrates,
inorganic salts, enzymes, and growth factors that are rich in
vitamin B complex.
Function: Source of growth factors, inorganic salts etc.
4. Yeast Extract
It contains proteins, amino acids, growth factors (Vitamin B),
Carbohydrates and inorganic salts like potassium and phosphates.
Function: Source of growth factors and hence excellent stimulators of
growth. It can be used as suitable for meat extract.

8. 5. Electrolyte
Mainly used are sodium chloride or other electrolytes.
Functions: Essential to maintain the osmotic pressure.
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6. Agar
Dried mucilaginous substance obtained from gelidium
species and other algae available as in powder form; contains
mainly long chain polysaccharides, protein like material and
inorganic salts.
Functions: it melts at 98°C and solidifies at 42°C, hence
used as solidifying agent.
7. Fermentable Compounds
Mainly used are sugars, alcohols etc.
Function: Act as source of energy, fermentation reactions
are helpful in the identification and classification of
organisms.
8. Buffers
Carbonates and phosphates are used as buffer.
Function: To resist change in pH of the
medium.

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11. Fermentation media
In most Industrial fermentation processes there are several stages, where media are
required.
1. Several inoculum (Starter culture) propagation steps,
2. Pilot-scale fermentation.
3. The main production fermentation.
The particular composition of a fermentation medium can be simple to complex. The
fermentation media can either be liquid, known as broth, or it can be a solid-state
fermentation. The media should satisfy all the nutritional requirements of the
microorganism and should also obtain the target molecule. A typical media requires a
carbon source, a nitrogen source, salts, water and micronutrients.
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12. A synthetic medium, is a medium in which all of the constituents are
specifically defined and known compounds. Every constituent is a
relatively pure compound, and the exact amounts incorporated into the
medium are known. Thus, an example of a synthetic medium would be
one containing inorganic salts, water, purified sugar, and an ammonium or
nitrate compound or an amino acid to supply combined nitrogen.
Advantages of the synthetic media:
✓Minimum Foaming in synthetic media.
✓Easy recovery and purification.
✓Individual components in the medium are easily deleted or
added.
✓Error due to media composition is held at a minimum.
✓may allow as to predict alternate substrates or other means for
further manipulation of the medium to increase yields.
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Synthetic Media

13. Crude Media
Crude media are
obtained from a range of
sources. Regardless of
the source, however, the
primary requirement is
that these nutrients be
inexpensive. In this
regard, the by-products
of agriculture have
always provided the
cheapest source of
medium components
and particularly of
carbon substrates.
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Components
found in
high amount
in
fermentation
media.
Water
Inorganic salts
Growth factors
Precursors
Buffers
Antifoam
Carbon source and nitrogen source
Oxidation-reduction potentials
Restricted nutritive levels.
Inhibitors

15. Water :
Water supports micro organism live in an aqueous environment in which
most of their nutritive requirments are dissolved .
Media usually contain more than 70% and often greater than 90% water.
The water supplies trace minerals, metals, such as copper, molybdenum,
zinc, boron and other substances that may be required in minute quantities
by micro organism.
The water used in media make-up also can affect the quality of the product.
Thus, as a medium component, the quality of the water can be critical.
Therefore, a supply of good cool water of constant composition is essential
to the fermentation industry.
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16. Inorganic nutrients :
Inorganic nutrients usually present little, a problem when crude media are
employed, because the common anions and cations occur insufficient quantities
such as phosphate, sulphate, magnesium, ammonium ions, it can be met by
adding potassium phosphate, magnesium sulphate and ammonium sulphate or
phosphate.
Calcium carbonate is sometimes added to neutralize acidic fermentation
products or to drastic decreases in pH which can occur during microbial growth.
Crude media provide excesses of toxic ions than to be deficient in required
ions.
Thus, it is possible that various components of the medium or the complete
medium may require a pre-treatment with ion exchange resins or some other
procedure to remove toxic quantities of certain metals such as copper, which can
poison sensitive microbial enzyme systems.
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17. Growth factors:
The growth-factor requirments of Industrial fermentation micro- organisms
are usually provided by the crude constituents of the fermentation medium..
Yeast products such as cell hydrolysates or autolysates are particularly the
sources of microbial growth factors if additional media sources are
required.
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18. Precursors:
• Precursors are defined as “substances added prior to or simultaneously with
the fermentation, which are incorporated without any major change into the
molecule of the fermentation product.
• Specific examples of the use of precursors are the addition to the medium
of phenylacetic acid as a precursor for penicillin-G and addition of
inorganic cobalt for production of vitamin-B12.
• It should be pointed out that the precursor can also dictate as to which
product from among several possibilities will be produced in greatest
amount.
• Thus, with cornsteep liquor in the penicillin fermentation up to six different
penicillins are possible products, but an addition of phenylacetic acid, the
major product becomes penicillin-G.
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19. Buffer:
Fermentation media contain buffers to retard gross changes in pH values during
microbial growth. The buffers can be specifically added for their buffering capacity
or they may be normal constituents of a medium, serving a dual function as both
buffer and nutritive source. During microbial growth, pH changes can occur for one
of several reasons.
Calcium carbonate often is incorporated in fermentation media to provide
neutralization of acidic fermentation products. It allows only slow reaction with
acidic products. Media containing considerable quantities of protein, peptides and
amino acids possess good buffer capacity in the pH range near neutrality. Additional
buffering capacity in this pH range also is provided by phosphates such as the
system of mono- and dihydrogen potassium or sodium phosphates. At lower, pH
values, acetic acid and other organic acids provide buffering capacity
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20. Antifoam:
• Antifoam agents of the crude types must be considered as potential
nutritive sources of alcohols and fatty acids for fermentation media.
❖Inert Antifoams.
❖Antifoam made from crude organic materials.
• In crude media , animal and vegetable oils are often used, for example,
lard oil, corn oil and soybean oil. Long-chain alcohols such as
octadecanol also are of value. For example, Lard oils mixed with
octadecanol for penicillin fermentations.
• The use of inert antifoam agents, such as various silicone compounds is
the ideal way to control foam, but these agents, are too expensive for use
in large-scale industrial fermentation.
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21. Oxidation-Reduction potential:
These agents usually present little problem in highly aerated fermentations,
but they must be considered in anaerobic fermentations.
Amino acids such as cysteine, peptides, such as glutathione and organic
acids such as thioglycolic acid will poise the medium at relatively low
oxidation –reduction potentials for growth of anaerobic or microaeroplilic
microorganisms.
In addition, the medium is heated and cooled just before inoculation to
drive out dissolved oxygen.
Certain media, such as cooked corn mash for the anaerobic clostridia,
further reduce oxygen penetration by their gel-like properties.
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22. Restricted Nutritive levels:
In the production of organic acids by fungi, the yields for other reasons, is
relatively poor. Also, the control of the combined nitrogen level of a
medium to retard microbial growth is beneficial in fermentations such as
that for fat production by the yeast Rhodotorula gracilis. Replacement
culture with fungal mycelium is another good example of the use of
restricted growth medium. A fungus, such as Aspergillus niger for gluconic
acid production, is first grown on a medium that provides good growth and
product formation , then the mycelium is separated from its medium and
placed in fresh medium high in carbon substrate (sugar),, but lacking
combined nitrogen so that additional growth cannot occur. Further
fermentation occurs enzymatically, in this case, by the glucose oxidase of
the mycelium.
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23. Industrial fermentations need raw materials that fulfill the requirements of
the organism (suitable carbon and nitrogen source, minerals and specific
nutrients) and that are available in a high quantity and quality. 23
Raw
materials in
industrial
fermentation
Molasses
Cellulose
Whey
Sulphite waste liquors
Wood hydrolysates
Rice straw
Corn steep liquor
soybean meal

24. Raw materials in Industrial fermentation
sources
Substrate
used as
carbon
source.
Molasses
Malt extract
Cellulose
Starch and
dextrins
Whey
Ethanol
Animal fats
Oils
Sulfite waste
liquors
Substrate
used as
nitrogen
source.
Corn steep liquor
Yeast extracts
Peptones
Soybean meal
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26. SACCHARINE RAW MATERIAL
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27. Molasses
• Molasses is a thick, sweet syrup, inevitably resulting from beet or cane sugar
production. Molasses contains unextractable sugar, vitamins and minerals,
such as calcium, sodium, potassium and magnesium. Molasses is an inevitable
final material arising from sugar production.
❑Types –
➢Sugar beet molasses.
➢blackstrap molasses
➢high-test or invert molasses
➢Hydrol molasses
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28. Beet molasses
Beet molasses are produced by the same process employed for cane molasses.
Vitamins such as biotin, pyridoxine, thiamine pantothenic acid and inositol are
present in beet molasses. Beet molasses have limited biotin
Therefore, in fermentation involving yeast culture, a small amount of cane
black strap molasses. Because yeast require for their growth.
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29. Blackstrap molasses
It is prepared from sugarcane normally is the cheapest and
the most used sugar source for industrial fermentations.
In the commercial production of sugar, the juice from
crushed sugarcane is concentrated to allow crystallization
of its sucrose.
The crystallized sugar is then separated from its mother
liquor, and the mother liquor is further concentrated to
allow recovery of additional crops of crystalline sugar the
mother liquor contain approximately 52% (30% sucrose
and 22% invert sugar).
It is particularly rich in pantothenic acid, biotin, thiamine,
phosphorous and sulphur. The organic nitrogen content is
less than beet molasses, since it does not contain betaine.
When this molasses is used as a fermentation medium
component, it is considered to contain 50% fermentable
sugars.

30. Invert molasses/ High-test molasses
The whole sugarcane juice is partially inverted to prevent sugar crystallization, the sugar
is partially hydrolyzed to Monosaccharides with heat and acid and then neutralized and
concentrated without removal of sugar.
contains approximately 70-75% sugar contains much of the original sugar of the cane
juice, although it has been partially hydrolyzed to D-glucose and D-fructose.
It is produced only during years of sugarcane overproduction.
Hydrol molasses
Hydrol is a molasses, resulting from manufacture of crystalline dextrose from corn
starch.
It contains 60% sugar.
High salt concentration, that must be considered if used in any fermentation process.
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31. whey
• Whey is the liquid remaining after milk has been curdled and strained. It
is a byproduct of the manufacturing of cheese or casein and has several
commercial uses. Sweet whey is a byproduct resulting from the
manufacture of rennet types of hard cheese, like cheddar or Swiss cheese.
Acid whey (also known as sour whey) is a byproduct brought out during
the making of acid types of dairy products, such as strained yogurt.
• Whey proteins consist of α-lactalbumin, β-lactoglobulin, serum albumin,
immunoglobulins, and proteose peptones.
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32. STARCHY RAW MATERIAL
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33. Cellulose
Cellulose is being extensively studied as a substrate for conservation to
sugar or alcohol.
The world’s annual cellulose production is estimated at 1011 metric tons;
much of it exists as waste, in such forms as straw, corn cobs, wood wastes,
peat, bagasse, and waste paper.
It is usually not possible to use cellulose directly as a carbon source, so it
must first be hydrolyzed chemically or enzymatically.
The sugar syrup formed from cellulose hydrolysis has been used for
ethanol fermentation, and the fermentative production of butanol, acetone
and isopropanol is also being considered.
Work is in progress to develop one-step processes for direct conversation
of cellulose to ethanol, using fermentative organisms which produce
cellulases.
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35. Sulphite waste liquor
It is a manufacture of paper pulp, wood is subjected
to hydrolysis which is brought about with the help of
calcium bisulfite under heat and pressure.
Sulfite waste liquor contains 10-12% solids, of which
sugars make up about 20%.
It contains sugar in the form of hexoses and
pentoses.It is used in the industrial production of
ethyl alcohol using Saccharomyces cerevisiae and in
the growth of Torula utilis cells for animal feed.
It contains free sulfur dioxide or sulfurous acid
which is toxic to microorganisms
These toxicants are removed by steam stripping or
precipitation with lime.
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36. Wood molasses (Acid Wood hydrolysates)
It is produced by acid hydrolysis of wood cellulose itself.
This may produce 65-85% fermentable sugars.
Sulphuric acid of about 0.5% concentration is used at a temperature range of
150 to 1850C. Using a continuous process a syrup is obtained from saw dust.
This syrup may contain 4 to 5% reducing sugars (a mixture of glucose and
pentoses) with an overall yield of 45 to 55%.
It is subjected to concentration to give a kind of wood molasses.
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37. Rice Straw
Rice straw and related agricultural materials can serve as a good
source of cellulose.
It is a poor quality animal feed in its natural state because of its
bulkiness, poor palatability, low protein content and low
digestibility.
Numerous microorganisms are capable of using cellulose for their
growth.
Rice straw has been used as a fermentation medium in the
production of silage and single cell protein (SCP), mushroom
cultivation etc.
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38. NITROGEN SOURCES
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39. Cornsteep liquor
Cornsteep liquor is the water extract by-product resulting from the
steeping of corn during the commercial production of corn starch, gluten,
and other corn products.
The used or spent steep waters are concentrated to approximately 50%
solids, and this concentrate, known as cornsteep liquor, is used in the
commercial manufacture of feedstuffs and as a medium adjunct in the
fermentation Industry.
It was first extensively employed in fermentation media for the
manufacture of penicillin.
Of the 50% solids of cornsteep liquor, approximately half is lactic acid.
The rest includes aminoacids, glucose and other reducing sugars, salts and
vitamins and precursors such as those for the penicillin molecule.
Thus, if the cornsteep liquor is supplying certain medium components (
such as a particular aminoacid, vitamin or precursor) at low but critical
levels, it may be necessary to determine the specific level of this
compound as it is present in each lot of cornsteep liquor that is to be used
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40. Soybean meal
The material left after removing oil from the soya bean seeds are called as soya
bean meal. Soya bean meal contains approximately 8% w/w nitrogen.
This differs from corn steep liquor, since soya bean meal is a much more
complex nitrogenous source than corn steep liquor, and therefore not readily
available to microbes.
The most important constituents of soybean flour are protein, fat and phosphorus.
This is used as a ingredient for fermentation media in the production of
streptomycin.
Soymeal, the residue from soybeans after the extraction of soybean oil, is a
complex substrate.
Analysis shows a protein content of 50%, a carbohydrate content of 30%
(sucrose, stach-yose, raffinose, arabinoglucan, arabinan, and acidic
polysaccharides), 1% residual fat, and 1.8% lecithin.
Soy meal is frequently used in antibiotic fermentations; catabolite regulation;
does not occur because of the slow catabolism of this complex mixture.

41. References
fermentation media & raw materials 41
ARTICLE :
Anannya Gupta. Raw materials in fermentation industries. The Torch. 2021. 2 (13).
WEBSITE MATERIALS :
Food Biotechnology Course material by K V Anand Raj
BOOKS :
Textbook on Fermentation Technology (Vol.1) by H A Modi
Textbook on Industrial Microbiology by A H Patel
Textbook on Industrial Microbiology by L. E Casida
A Textbook of Industrial Microbiology by Wulf Crueger and Anneliese Crueger
Principles of Fermentation Technology by Stanbury and Whitaker, 2ed.

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44. • Optimal Mindset.
• Communication Skills.
• Level of Confidence.
• Gained Subject Knowledge Better.
• Presentation Skills.

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THANKS TO
THE CHAIRMAN.
THE SECRETARY.
MANAGEMENT COMMITTEE.
THE PRINCIPLE, SRI PARAMAKALYANI COLLEGE.
THE HEAD, DEPARTMENT OF MICROBIOLOGY.
THE STAFF MEMBERS, DEPARTMENT OF MICROBIOLOGY.