SECONDARY METABOLITE PRODUCTION

1. SECONDARY METABOLITE
PRODUCTION

2. INTRODUCTION
 Higher plants synthesize a wide variety of low
molecular weight compounds in addition to the
essential primary metabolites. These are called
secondary metabolites
 These secondary metabolites offer protection against
pests,they act as attractatnts and as the plant’s own
hormones

3. WHY SECONDARY METABOLITES
 Chemically warfare to protect plants from the attacks
by predators, pathogens, or competitors
 Attract pollinators are seed dispersal agent
 Important for abiotic stress
 Medicine
 Industrial additives

4. CLASSIFICATION OF SECONDARY
METABOLITES
 They are classified on the basis of
 1)chemical structure
 2)chemical composition
 3)solubility in various solvents
 4)pathways by which they are synthsized

5.  A simple classification of secondary metabolites
includes three main groups
 1)the terpenes- made from mevalonic acid,
composed almost entirely of carbon and hygrogen
 2) phenolics – made from simple sugars,
containing benzene rings, hydrogen and oxygen
 3) nitrogen-containing compounds – extremely
diverse, may also contain sulphur

6. ALKALOIDS
 Most are derived from a few common amino acids
(i.e..tyrosine , tryptophan , ornithine or argenine and
lysine
 Compounds have a ring structure and a nitrogen
residue
 Indole alkaloids is the largest group in this family ,
derived from tryptophan
 Widely used as medicine

7. CHARACTERISTICS
 They are bitter in taste.
 Derived from amino acids.The amino acids that are most
often serve as alkaloidal precursors are: phenylalanine,
tyrosine, tryptophan, histidine, anthranilic acid,
lysine and ornithine.
 Alkaloids form double salts with compounds of mercury,
gold, platinum and other heavy metals. These salts are
obtained as precipitate which are microcrystals.

8.  Insoluble or sparingly soluble in water, but the salts
formed on reaction with acids are usually freely
soluble.
 Most are crystalline solids although a few are
amorphous.

9.  Free alkaloids are usually soluble in polar solvents
like ether, chloroform
 Some alkaloids are liquid because of lacking of
oxygen in their molecules. (e.g coniine, nicotine,
spartenine)

10. Sources and Occurrence of Alkaloids
 Alkaloids can occur in plant kingdoms; among the
angiosperms,
 Leguminosae,
 Papaveraceae,
 Ranunculaceae,
 Rubiaceae,
 Solanaceae,
 Berberidaceae are outstanding alkaloid-yielding plants.

11. Uses of Alkaloids in Plants:
 Poisonous agents which protect plants against insects and
herbivores
 End products of detoxification reactions representing a
metabolic locking-up of compounds otherwise harmful to the
plants.
 For regulatory growth factors
 Reserve substance capable of supplying nitrogen or other
elements necessary to the plant’s economy

12. TERPENES
 Terpenes are natural products that are structurally
related to isoprene
 Isopropene
 (2-methyl-1,3-butadiene)
H2C C
CH3
CH CH2
OR

13. 1. The number of C atoms is a multiple of 5, C5
C10 C15 C20 C25 C30 C35 C40
2. Each group of 5 C is an isoprene subunit
3. They can be saturated or unsaturated
4. Many contain O atoms as well.

14. PHENOLICS
• Derived from aromatic amino acids, such as phenylalanine,
tyrosin, and trytophan.
• All contain structures derived from phenol
• Some examples:
 Coumarins: antimicrobial agents, feeding deterrents, and
germination inhibitors.
 Lignin: abundant in secondary cell wall, rigid and resistant to
extraction or many degradation reagents
 Anthocyanins
 Flavones
 Flavnols
 Phenols are present in every plant they attract pollinators to the
plant and even impact how these plants act with one another.
 .

15. GLYCOSIDES
• Compounds that contain a carbonhydrate and a
noncarbohydrate
• Glycosides are present in vacuoles in inactive form
• Glucosinolates: found primarily in the mustard family to
give the pungent taste
 There are four type of linkages present between glycone and
aglycone:
 C-linkage/glycosidic bond,
 O-linkage/glycosidic bond
 N-linkage/glycosidic bond
 S-linkage/glycosidic bond

16. Sources and effects of major
secondary metabolites
carotenoids are
organic pigments
occuring in plants
and are mostly
found in red orange
and yellow fruits
and vegetables .
they have
antioxidative effects
and prevents cancer
They boost the
immune system
and reduce the risk
of heart attacks

17. PHYTOSTEROLS
Found in-
sunflower seeds
,sesame,nuts and
soy beans
Protect against
colon cancerand
low cholestrol level

18. SAPONINS
Flavour additives
found in- legumes
and spinach
Boost the immune
system
Lower the
cholestrol level in
blood
Reduce the risk of
intestinal cancer

19. FLAVONOIDS
Organic pigment
Gives plant red
violet or blue colour
Inhibit the growth
of bacteria and
viruses
Protect the cells
against the damage
of freeradicals
Influence blood
coagulation

20. SULPHIDES
Contain sulphur
Belongs to the lily
family onions
,leeks, asparagus,
garlic
Inhibit the growth
of bacteria
Lower cholestrol
levels
Preventive efect
against cancer

21. CURCUMIN
•Extracted from
turmeric.
•Coloring and
medicinal uses
•Wound healing
•Antiulcer
•Anti inflammatory
•Antimicrobial &
antiviral
•Hepatoprotective
•Antioxidant
•No toxicity.

22. FLAVORS
Cocoa seeds
Vennila
Strawberry
Cinnamon
Cardamom
Cloves
Pepper
areca

23. PRODUCTION OF SECONDARY
METABOLITES IN CULTURES/BIOREACTORS
 1)choice of explants
 any part of the plant can be used
 dicotelydons are rather amenable for callus
tissue induction
 procured explants are sterilized prior to
culture by using ethanol

24. 2)NUTIENT CULTURE MEDIA
 To induce callus from an explant and to cultivate the callus
and cells in suspension

25. 3)Callus induction
 A callus cell culture is usually sustained on gel
medium. Callus induction medium consists of agar
and a mixture of macronutrients and
micronutrients for the given cell type. There are
several types of basal salt mixtures used in plant
tissue culture, but most notably modified Murashige
and Skoog medium White's medium, and woody
plant medium. Vitamins are also provided to
enhance growth such as Gamborg B5 vitamins. For
plant cells, enrichment with nitrogen phosphorus,
and potassium is especially important.

26. SUSPENSION CULTURE
 The growth rate of suspension cultured cell is
generally faster than that of the solid culture. The
former is more desirable, particularly in the
production of useful metabolites on a large scale

27. FERMENTORS OR BIOREACTORS
 Carboy system-described by tulecke and nickell in
1959,consists of a rubber-stoppered 20 litre carboy
fitted with four tubes. Filtered compressed air is
employed for oxygen supply, aeration and agitation
of the medium
 Stirred jar fermetor- used for microbial cultivation
 Agitation speed of 50 to 100 rpm

29. HAIRY ROOT CULTURE
 It is the culture produced after the infection of
explants and cultures by the gram negative soil
bacteria Agrobacterium rhizogens
 This process take advantage of the naturally occuring
hairy root disease in dicotyledons

30.  Agrobacterium recognizes some signal
molecules exuded by wonded plant cells and
becomes attached to it.
 The bacteria contain the Root inducing
plasmid ( Ri-plasmid).
 The bacteria Genetically trasfer part of the Ri-
plasmid called the trasfer DNA (T-DNA) to the
plant genome.

31.  The plant cell to proliferate by increasing the rate
of cell division ( cytokine expression ) and cell
elongation (auxin expression) to produce the hairy
roots.
 Produce the opines which is a type of unusual
amino acids ( octopine, agropine, nopaline,
mannopine amd cucumopine) which is used by the
bacterium as a carbon , nitrogen and energy
source.

32. Induction of hairy root cultures In Vitro
 Explants are wounded and then inoculated with
Agrobacterium rhizogenes.
 Usually two or three days later, the explant can be
transferred into solid media with antibiotics, such
as cefotaxime , vancomycine or ampicillin to kill or
eliminate redundant bacteria.
 The hairy roots will be induced within a short
period of time, which varies from one week to over
a month depending on different plant species.

33.  The decontaminated hairy roots can be
subcultured on phytohormone –free medium.

35. Commercial production of secondary metabolites
 higher plants contain a variety of metabolic products which
are useful as medicines ,food additives,perfumes,etc.
however , non-compromising reduction in plant
sources,increases in labour cost and other problems in
order to obtain these high value added substances from
natural plants ,have forced to develop analternative
approachtowards the use of plant cell culture for
production of these metabolic products .since ptc is not
affected by changes in environmental conditions such as
climate or natural depredation, improved production is
possible

36. PHARMACEUTICALLY
IMPORTANT CHEMICALS
1)Morphine
alkaloids
Codeine is an
anlagesic and
cough suppressing
drug from Papaver
somniferum
also morphine
can be converted
into codeine
many workers
have tried to
produce codeine by
undifferentiated
cells of these plants

37. BERBERINE
it is an
isoquinoline
alkaloid which is
produced and
distibuted in
roots of Coptis
japonica and
cortex of
Phellonderon
amurense
Berberine
chloride is used in
treatment of
intestinal
disorders

38. TORPANE ALKALOIDS
Scopolamine and
hyoscyamine areused
commercially as
anesthetic and
antispasmodic drugs.
Occur in the
leaves of members of
the family
Solanaceace including
Duboisia
myoporoides Datura
leichhartdtti,
Scopolia, Atropa,and
Hyoscyamus

39. ANTITUMOUR COMPOUNDS

40. VINCA ALKALOIDS
Extracted from
Catharanthus
roseus
Treat leukemias,
hodgkin’s
disease and
antitumor
activity

41. GINSENG
Roots of Panax
ginseng is
widely used as
tonic
Effective for
genetic
disorders,
diabetes and
weak circulation
Root contains
saponins and
sapogeninis

42. FOOD COLORING AGENTS

43. SAFFRON
Made from the
stamens of Crocus
sativus and used as
flavouring agent
and colourant
Plant contains
crocin(yellow
pigment),
safranal(fragrance),
picocrocin(bitter
substance)
Baked goods, soaps,
cheese
Medicinal value for
stomach ailments

44. SAFFLOWER YELLOW
Yellow pigment
obtained from the
floret of the
Carthamus
tinctorius
Major pigment is
carthamin
Carthamin is a
quinoid form of
isocarthamin
Used in baked
goods and
beverages

45. CHEWING GUMS CHICLE
Chicle a raw material
is obtained from the
latex of Achras
sapota
Chicle contains 60%
resin and 15% of
rubber
Resin consisits of
lupeo,α-amyrin andβ-
amyrin
Rubber consists of cis
and trans- 1,4-
polyisoprene
Synthesized through
mevalonic acid
pathway

46. TAXOL
It is a ditrpene
amide
Extracted from
the bark of wild
grown Taxus
brevifolia
Treats breast
and lung cancer