THE FERMENTATION PROCESS AND ITS TYPES ARE DISCUSSED HERE, WITH SOME EXAMPLES AND SYNTHESIS FORMED BY FERMENTATIONSUCH AS ANTIBIOTICS INCUDING PENICILLIN, STREPTOMYCIN AND VITAMINS A VITAMIN B2, VITAMIN B12.

1. FERMENTATION

2. CONTENTS
 Introduction
 Types of fermentation
 Production of –
1. Antibiotics : Penicillin & Streptomycin
2. Vitamins: B2 & B12
 References
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3. Introduction
 Pasteur definition: “life without air”, anaerobe red
ox 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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4. 4
Some important fermentation products
PRODUCT ORGANISM USES
Ethanol Saccharomyces
cerevisiae
Industrial
solvents
beverages
Glycerol Saccharomyces
cerevisiae
Production of
explosives
Lactic acid Lactobacillus
bulgaricus
Food and
pharmaceutical
Acetone &
Butanol
Clostridium
acetobutylicum
solvents

5. Difference between FERMENTATION
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AEROBIC
FERMENTATION
ANAEROBIC
FERMENTATION
Set of chemical reactions
involved in production of
energy by completely
oxidizing food.
Chemical breakdown of
organic substrate into
ethanol or lactic acid by
micro-organism in absence
of oxygen
Occurs in both cytoplasm
and mitochondria
Occurs in cytoplasm
Occurs in higher animals
and plants
Occurs in yeast ,parasites,
and bacteria

6. 6
Uses molecular oxygen as
final electron acceptor in
ETC
Does not use oxygen
Produces 6 water
molecules/glucose
Does not produce water
Glucose completely broken
into CO2 & O2
Glucose incompletely
oxidized either into ethanol
& lactic acid
NAD+ regeneration occurs
in ETC
NAD+ regeneration occurs
during partial oxidation of
Pyruvate
36 ATP is produced Only 2 ATP is produced
Continued……

7. Penicillin
 Also known as Penicillium Notatum / Chrysogenum.
 It is common in temperate and subtropical region and
can be found on salted food products, but it is mostly
found in indoor enviroments, especially in damp or
water damaged buildings.
 It is source of several β-lactum antibiotics, which
inhibits biosynthesis of bacterial cell wall.
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8. Beta lactum ring
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9. Production
 P. chyrosegenum is high yielding strain and therefore
most widely used as production strain.
Innoculum preparation- various sequential steps are
necessary like:
1. A starter culture is needed for innoculation.
2. After getting growth on solid media, one or two
growth stages should allowed in shaken flask
cultures to create a suspension, which can be
transfered to seed tanks for further growth.
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10. 3. After about 24-28 hours, the content of seed tanks is
transfered to primary fermentation tanks.
4. All the bio-parameters like temperature, pH, aeration,
agitation,etc. should be properly maintained.
A. BIO-PARAMETERS-
 pH- near 6.5
 Temperature – 260- 280 C
 Aeration- Continous stream of sterilized air is pumped
into it.
 Agitation- have baffles which allows constant agitation
(200rpm).
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11. B. NUTRIENT MEDIA
 Fermentation broth contains all necessary elements
required for the proliferation of micro-organisms.
 Generally it contains-
1. Carbon source
2. Nitrogen source
3. Mineral source
4. Precorsors
5. Anti – foaming agents
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Composition of nutrient media for penicillin
1. Corn steep liquor 3.5%
2. Lactose 3.5%
3. Glucose 1%
4. Calcium carbonate 1%
5. Potassium hydrogen phosphate 0.4%
6. Edible oil 0.25%
Phenyl acetic acid as preservative

13. C. RECOVERY
 The recovery of Penicillin is carried out in three
successive stages:
1. Removal of mycelium
2. Counter current solvent extraction of penicillin
3. Treatment of crude extract
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14. For increasing yield of penicillin
 Adding corn syrup
 Selection of strain
 Mutation and selection
 Sexual reproduction
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15. streptomycin
 Streptomycin is an bactericidal antibiotic drug
belonging to class of aminoglycosides.
 Derived from actinobacterium Streptomyces griseus.
 Used against gram negative bacteria especially.
 Chemical composition – It contains 3 sugars derived
from glucose with C, N, O & H elements.
 Chemical formula- C21H39N7O12
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16. INNOCULUM
 Streptomyces griseus maintained in soil stocks or
lyophilized in carrrier are innoculated into sporulation
medium which builds up mycelial innoculum.
THE HOCKENHUL MEDIUM
1. Glucose 2.5%
2. Extracted soya meal 4%
3. Distillers dried soluble 0.5%
4. Sodium chloride 0.25%
 pH 7.3-7.5
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Production

17. A. NUTRIENT MEDIA
 Medium is a nutritive substance in which cultures
are grown for scientific purposes.
 The culture medium for streptomycin consists of:
1. Carbon source
2. Nitrogen source
3. Animal oils
4. Vegetable oils
5. Mineral oils
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18. B. FERMENTATION PROCESS
 Spores of S. griseus are innoculated into medium to
establish a culture with high mycelial biomass for
introduction into innoculum tank, using innoculum to
initiate the fermentation process.
 Yield in production vessel responds to high aeration &
agitation condition
 Other condition are:
Temperature 250-300C
pH range 7-8
Time- 5-7 days
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19.  The fermentation process for production of
streptomycin involves three phases :
1. Phase I (last up 24 hours)
2. Phase II (takeplace5-7 days)
3. Phase III (harvested before cell lysis)
C. RECOVERY & PURIFICATION
1. Removal of mycelium
2. Recovery process
3. Purification
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20. Cyanocobalamine ( B12)
 Cyanocobalamin is industrially produced stable cobalmin
from which is not found in nature
 It is obtained exclusively by fermentation process.
 Merck began production of vitamin B12 by Pseudomonas
dentrificans in 1952 & have improved the efficiency of
culture more than 30 folds relative to the performance of
original soil isolates by genetic manipulations and
microbial screening.
 Mutagenic treatments have resulted in improved activity.
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21.  Mutagenic treatments have resulted in improved
activity.
 Structure of cyanocobalamine :
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Downstream process of vitamin B12

23. Riboflavin (B2)
 Riboflavin has been produced commercially by chemical
synthesis, by fermentation and by combination of
fermentation and chemical synthesis.
 Fermentation route has been widely used as it produces
the vitamin in single step, resulting in substantial cost
savings.
 Most of the producers like BASF, Roche, ADM/
Aventis, Hubei Gaungii prefer fermentative production.
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24.  Although bacteria (Clostridium species) & yeasts
(Candida species) are good producers, two closely
related Ascomycete fungi, Eremothecium ashbyii
& Ashbya gossypii, are considered the best
riboflavin producers.
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