Vitamins are organic compounds that are essential in small amounts for normal metabolism and good health. They are produced naturally by plants and microorganisms or synthetically. The document discusses the production of several vitamins (B2, B6, B12, C) through microbial fermentation. Optimal conditions like pH, temperature and carbon/nitrogen sources influence microbial vitamin production. Strain improvement through mutagenesis, genetic engineering and culture optimization has increased industrial vitamin yields.

1. PRODUCTION OF

2. INTRODUCTION

3. INTRODUCTION
● Vitamins are a group of organic food substances or nutrients
found only in living things, plants and animals or produced
synthetically in a laboratory so can be ingested in their pure
form as nutritional supplements.
● Most commercial vitamins are made from synthetic vitamins
which are cheaper and easier to produce than natural
derivative
● Vitamins are necessary in small amounts for normal
metabolism and good health.
● The absence of certain vitamins can cause disease, poor
growth, and a variety of syndromes.
The vitamins are named by letters—vitamin A, vitamin C, D, E,
K, and the group of B vitamins.

4. ● Vitamins were discovered by Dutch physician, Christian
Eijkmann, who won the Nobel Prize in physiology and medicine
in 1929.
● Vitamins make it possible for other nutrients to be digested,
absorbed and metabolized by the body.
● Vitamins have diverse biochemical functions, including
function as hormones (e.g. vitamin D),antioxidants (e.g.
vitamin E), and mediators of cell signaling and regulators of cell
and tissue growth and differentiation (e.g. vitamin A)
● The largest number of vitamins (e.g. B complex vitamins)
functions as precursors for enzyme cofactor bio-molecules
(coenzymes) that act as catalysts and substrates in metabolism.
● Vitamins are divided into two classes based on their solubility:

5. fat-soluble vitamins water-soluble vitamins
vitamin D Vitamin B 12 (cyanocobalamin)
vitamin E vitamin B9 (folate or folic acid)
vitamin A vitamin B7 (biotin)
vitamin K vitamin B6 (pyridoxine)
Vitamin B5 (pantothenic acid)
vitamin B3 (niacin)
vitamin B2 (riboflavin) ,vitamin
B1(thiamin)vitamin C (ascorbic
acid)

6. Sources of vitamins
VitaminS SOURCES
vitamin A vegetable
vitamin D milk, fatty fish,sunlight
vitamin K broccoli, Brussels sprouts, cabbage,
cauliflower, kale, spinach and soybeans
Thiamine (vitamin B1) fortified breads, cereals, pasta, whole
grains, lean meats (especially pork),
fish,dried beans, peas and soybeans.
riboflavin(B2) organ meats (liver, kidney and heart)
almonds, mushrooms, whole grain,
soybeans and green leafy vegetables
Niacin(B3) dairy products, poultry, fish, lean
meats, nuts and eggs.
Most of the vitamins can be found in plant and animal
sources. They can also be chemically synthesized.

8. vitamin B6 white meat (poultry and fish),
bananas, liver, whole-grain breads and
cereals, soyabeans and vegetables.
folic acid(B9). Beans, leafy green vegetables, citrus
fruits,beets, wheat germ, and meat
biotin organ meats, oatmeal, egg yolk, soy,
mushrooms, bananas, peanuts, and
brewer s yeast.
vitamin C Cabbage and many dark green leafy
vegetables
pantothenic acid(B5) cheese, corn, eggs, liver, meats,
peanuts, peas, soybeans, brewers
yeast and wheat germ.

9. MICROBIAL PRODUCTION
OF VITAMINS

10. ● All vitamins can be extracted from natural
sources, but fat-soluble vitamins are often
produced commercially by synthetic processes.
● Several vitamins are now industrially produced
and are used in foods, pharmaceuticals and
cosmetics.
● Presently, few of the vitamins are exclusively
produced via chemical synthesis, while a few
others are produced either by chemical synthesis
or via extraction processes.
● Microbial production is commercially feasible for
some vitamins, such as vitamin B2,vitamin B6,
vitamin B12 and vitamin C.

11. ● Fermentation technologies provide an
alternative to chemical processes in
the production of wide range vitamins
from microbes.
● Different methods like media
optimization, mutation and screening,
genetic engineering and biocatalyst
conversion have been used for
improvement of the production of
vitamins.

12. VITAMIN B2 OR
RIBOFLAVIN● vitamin B2 or riboflavin functions as part of
metabolic systems concerned with the oxidation of
carbohydrates and amino acids. It is active not in
the free form but in more complex compounds
known as coenzymes, such as flavin
mononucleotide (FMN) and flavin adenine
dinucleotide (FAD), or flavoprotein.
● It plays a key role in energy metabolism, and is
required for the metabolism of fats, ketone bodies,
carbohydrates and proteins.
● Various biotechnological processes have been
developed for industrial scale riboflavin
biosynthesis using different microorganisms,
including filamentous fungi such as Ashbya
gossypii, Candida famata as well as the bacteria
B.subtilis.

13. ▪ Majority of the
microorganisms can
synthesize riboflavin
from simple medium
components. Some
bacteria, yeast and
yeast-like fungi are able
to overproduce
riboflavin under specific
conditions.
▪ Food Sources for
Riboflavin: Sources
include liver, eggs, dark
green vegetables,
legumes, whole and
enriched grain products,
and milk.

14. VITAMIN B6
● vitamin 6 is a water-soluble vitamin and includes
a group of closely related compounds pyridoxine
(PN), pyridoxal (PL), and pyridoxamine (PM).
● Pyridoxal phosphate (PLP) is the active form and
function as cofactor in many reactions of amino
acid metabolism, including transamination,
deamination and decarboxylation.
● Pyridoxal phosphate is involved in
macronutrient metabolism, neurotransmitter
synthesis, histamine synthesis, hemoglobin
synthesis and gene expression.

15. ● Major sources of vitamin
B6 include cereal grains,
legumes, vegetables,
potatoes, milk, cheese,
eggs, fish,liver, meat and
flour.
● Several strains such as
Klebsiella, Achromobacter
cycloclastes,
Flavobacterium, Bacillus,
Pichia guilliermondii and
Rhizobium were reported
to produce vitamin B6.

16. VITAMIN B-12

17. INTRODUCTION:
● Vitamin B12, also
called cobalamin, is a water-
soluble vitamin.
● It is one of eight B vitamins.
● It is involved in
the metabolism of every cell of
the human body.
● No fungi, plants, or animals
(including humans) are capable
of producing vitamin B12.
● B12 is the largest and most
structurally complicated vitamin
and is produced industrially only
through bacterial fermentation.
● Vitamin B12 was discovered as a
result of its relationship to the
disease pernicious anaemia.

18. SOURCES
● ANIMAL SOURCES:
Liver, muscles, eggs, milk,
meat, insects, crab meat,
lamb, fish eggs,
Clams, mackerel etc.
● MICROBIAL SOURCES:
B12 is produced in nature
only by some prokaryotes.
It is synthesized by
some gut bacteria in
humans and other animals.
Faeces are a rich source of
vitamin B12 and many
species, including dogs and
cats, eat feces.

19. PRODUCTION:
● The complete
laboratory synthesis of B12
was achieved by Robert
Burns Woodward and Albert
Eschenmoser in 1972.
● Industrial production of
B12 is achieved
through fermentation of
selected microorganisms.
● The species Pseudomonas
denitrificans and Propioniba
cterium
freudenreichii subsp. Sherm
anii are more commonly
used today.
● Carried out under anaerobic
conditions in the absence of
precursor.
● It is manufactured by

20. VITAMIN C

21. INTRODUCTION:
● Vitamin C is also known as
Ascorbic acid and L-ascorbic
acid.
● The first vitamin to be chemically
produced
● It is used as a dietary
supplement.
● Vitamin C is an essential nutrient
involved in the repair of tissue
and the enzymatic production of
certain neurotransmitters.
● It also functions as an
antioxidant.
● Foods containing vitamin C
include citrus fruits, broccoli,
Brussels sprouts, raw bell
peppers, and strawberries.
● Historically, vitamin C was
used for preventing and
treating scurvy.

22. SOURCES

23. PRODUCTION:
● Vitamin C is produced
from glucose by two main routes.
● The Reichstein process,
developed in the 1930s, uses a
single pre-fermentation followed
by a purely chemical route.
● And the modern two-
step fermentation process,
originally developed in China in
the 1960s.
● Both processes yield
approximately 60% vitamin C
from the glucose feed.

25. PARAMETERS
CONTROLLING VITAMIN
PRODUCTION BY
FERMENTATION
:

26. Several physical and nutritional parameters influence vitamin
production:
❑ Physical parameters:
▪PH
The maximum biosynthesis of vitamin B12 recorded at pH 10 by
using mixed cultures from Streptomyces halstedii and Bacillus firmus.
Kolonne et al (1994) reported pH of the medium influence riboflavin
production. Highest yields were obtained at constant pH of 4.5 and
5.5, while little or no riboflavin was detected at either pH 3.5 or
8.5. when the pH was raised from 6.0 to 6.5, the biomass as
well as the riboflavin production was enhanced. An increase in pH
from 6.5 to 7.0 did not repress riboflavin production but inhibited
biomass production drastically, indicating pH6.5 to be more
conducive for all the enzyme activities involved in the growth of
the fungus. The pH 7.0 did not promote growth or repress or
derepress riboflavin production, indicating that the enzymes
involved in flavinogenesis were stable and active in abroader pH

27. ▪Temperature
A decrease in fermentation temperature from 32
to 24°C led to a decrease in vitamin B12 formation.
optimal vitamin B12 production required a temperature
of 40°C and aerobic conditions (0.5 vvm aeration at
100 rpm) with a pH value of 6.5.
Different Incubation Temperature:
The maximum biosynthesis OF vitamin B12 could be
recorded within an incubation temperature of 35°C for
mixed cultures of Streptomyces halstedii and Bacillus
firmus .
▪Incubation Period
The maximum 12biosynthesis of vitamin B was recorded
with an incubation period 9hrs. for mixed cultures of
Streptomyces halstedii and Bacillus firmus.

28. ▪Different Inoculum Size
The maximum vitamin B biosynthesis
was obtained in the presence of 6 discs by using mixed
cultures from Streptomyces halstedii and Bacillus firmus.
❑ Nutritional parameters:
▪ Carbon Source
The galactose is the best carbon source for biosynthesis
of vitamin B12.
▪Nitrogen Sources
The L-asparagin is the best nitrogen for the
biosynthesis of vitamin B12 followed by sodium nitrate,
ammonium chloride, glycin, potassium nitrate,
ammonium sulphate, glutamic cid, argnine, beef extract
and peptone.

29. STRAIN IMPROVEMENT FOR
VITAMIN PRODUCTION

30. Recent advances in the cloning of riboflavin,
cobalamin and biotin biosynthesis genes in
Bacillus species provide potential for the use of
Bacillus strains in vitamin production.
RIBOFLAVIN (vitamin B2)
It was observed that mutations in the ribO
operator site of the riboflavin operon, which
contains six structural genes, resulted in the
development of a number of overproducers.
The strategy to develop a commercial riboflavin
production strain of Bacillus subtilis was to
combine classical genetic mutant selection and
fermentation improvement with genetic
engineering of the riboflavin biosynthetic genes.

31. ● Perkins et al (1999) developed a recombinant
strain of Bacillus subtilis that produces
commercially attractive levels of riboflavin.
The B. subtilis riboflavin production strain
contains multiple copies of a modified B.
subtilis riboflavin biosynthetic operon.
● Park et al (2007) mutated Ashbya gossypii
spores by exposure to UV light. The mutant
ZP4 strain, produced threefold riboflavin than
that of the wild-type.
● Koizumi et al (2000) carried out strain
improvement studies for the production of
riboflavin (vitamin B2) through metabolic
engineering using recombinant DNA
techniques in Corynebacterium
ammoniagenes. The recombinant strain
produced 17-fold as much riboflavin as the
host strain.

32. • Hans et al (2008) reported overexpression of
riboflavin biosynthetic pathway by sequential
deregulation of all the genes, by exchange of
their native promoter with the strong and
constitutive glyceraldehyde-3-phosphate
dehydrogenase promoter (PGAP).

33. L-ASCORBIC ACID (vitamin C)
•Running et al (1994) described a one-step
fermentation process for the production of L-
ascorbic acid using the heterotrophic green
microalga Chlorella pyrenoidosa.
•Whereas low amounts of up to 40 mg/l L-
ascorbic acid were obtained by culturing wild-
type cells, after repeated rounds of chemical
mutagenesis and fermentation optimization,
improved amounts of up to 2 g/l L-ascorbic acid
were achieved. The produced amounts
correspond to a 70-fold improvement of L-
ascorbic acid formation as compared to the
wild-type.

34. •Because the bulk of L-ascorbic acid produced by
the microalgae remained intracellular, the process
was patented as a method for the production of L-
ascorbic acid enriched biomass for animal feed or
as dietary supplement.
•Shibata et al (2000) generated a recombinant
Pseudomonas putida strain in which the genes
encoding D-sorbitol dehydrogenase, L-sorbose
dehydrogenase, and L-sorbosone dehydrogenase
from G. oxydans were introduced and expressed.
After optimization of culture conditions, the
recombinant strain produced 16 g/l 2-keto-L-
gluconic acid from 50 g/l d-sorbitol.