Plants provide us everything, do we? Here the production of alkaloids, its uses, and many more important aspects are discussed. See and share https://www.linkedin.com/in/shradheya-r-r-gupta-54492984/

1. PLANT SECONDARY
METABOLITES
PRODUCTION OFALKALOIDS
Name :- Shradheya R.R.
Gupta
Course :- B. Sc.
Biotechnology 2nd year
Year :- 2014- 15
Submission to :- Dr. Parul

2. CONTENT
Introduction
Important alkaloids of plant origin
Different alkaloids market value
Why tissue culture to produce alkaloids?
Strategies for production
Accumulation of alkaloids
Site of alkaloid accumulation
Uptake and storage
Permealization
Extraction of alkaloids
New discoveries
Conclusion
Prospects

3. Introduction
 Secondary metabolites are organic molecules that are not
involved in the normal growth and development of an
organism.
 Alkaloids are those basic substances which contain one or
more nitrogen atoms, usually in combination as part of a
cyclic system.
 Alkaloids have been known to man for several centuries and
are used for human welfare.
 Of all known natural products, about 20% are classified as
alkaloids, but only 30 alkaloids are produced at the
commercial level.

4. Important Alkaloids of Plant Origin
Alkaloid Source Property
Atropine Atropa belladonna Anticholinergic
Berberine (a) Berberis spp. (a) Antispasmodic
(b) Coptis japonica (b) Antiprotozoal
Codeine Papaver somniferum Sedative, analgesic
Colchicine Colchicum autumnale Antimitotic
Caffeine (a) Coffea arabica
(b) Camellia sinensis
Stimulant
Morphine Papaver somniferum Analgesic, sedative
Quinine Cinchona ledgeriana Antimalaria
Vinblastine Catharanthus roseus Anticancer

5. Different Alkaloids Market Value
Alkaloid Culture type Yield in
culture
Price (US $/g)
Quinine Cell suspension Traces 0.50
Morphine Cell suspension 0.25 g/l 340.00
Camptothecine Cell suspension 0.00025% DW 480.00
Elliptine Cell suspension 0.005% DW 2630.00
Vinblastine Shoot culture Traces 10,530.00
Vincristine Shoot culture Traces 25,200.00

6. Why tissue culture to produce
alkaloids?
• Most of the secondary metabolites are
accumulated after a certain age of maturity of the
plant(e.g. Cinchona, Rauwolfia, Camptotheca,
Ochrosia).
• It is difficult to increase the area under plantation.
• Harvesting of plants from natural forest resources
is difficult, but also make them endangered(e.g.
Ephedra gerardiana).
• World political scenario also affect the supply of a

7. Strategies for Production
• Culture produce very low amount of secondary
metabolites compared to intact plant.
• Callus culture are slow growing system and accumulate
more compared to fast growing cell suspension culture
but, cell suspension culture is important for industrial
production.
• Optimization of alkaloid production is done using
physical factors, nutrients, plant growth regulator.
• Cultures are mixture of producer and non producer cells
and selection of high alkaloid producing cells enhance
the production. But these high alkaloid producing cells
are not stable and repetitive clonal selection is required

8. Different studies have been conducted, some are:-
1. Zenk et al. 1977:-
• Used 2 step culture system, medium for optimal
growth and medium for production of alkaloids.
In the latter growth is arrested by manipulating
the nutrients.
2. By Rideau 1987:-
• Fast growing cultures accumulate alkaloid in low
amount during the exponential phase of growth
and in high amount during the stationary phase.
• During this phase nutrients are exhausted and
primary metabolites is diverted to the synthesis

9. 3. Robins et al 1991:-
• Learn the mechanism of transport and
accumulation , enzymes involved in the
synthesis of alkaloids, identification of genes
and transfer of such genes for expression in
other eukaryotes or prokaryotes.
• These strategies to improve the production of
alkaloids and secondary metabolites in general.

10. Accumulation of Alkaloids
 Site of alkaloid production:-
• Product may be accumulated in some specialized
glands, trichromes, tissues, organ or seeds,
information is not complete regarding the site of
biosynthesis and accumulation. Accumulation of a
compound in a specific site does not necessarily
mean that it has been synthesized in the self same
tissue. e.g.- Lupin synthesized in leaf and
transport to epidermal via the phloem. Ajmalicine,
harmane and nicotine synthesized in roots.
• Availability can be investigated on the bases of

11.  Uptake and storage :-
• When berberine was added to various plant cell
cultures, it showed strong cytotoxicity to
berberine-non-producing plant species like
tobacco.
• C. japonica and T minus cells have an ability to
take up berberine from the medium against the
concentration gradient when added to the culture
medium and the absorbed berberine was
exclusively accumulated in the vacuoles (mainly
of rhizome).
• Two transport events are involved, i.e., uptake of
berberine at the plasma membrane(done by ABC

12. • CjMDR1 receptor was localized to the plasma
membrane of C. japonica, this accumulation is done
in the cells of rhizome.
• Since the rhizome is the sink organ also for starch,
this plant accumulates the alkaloid having strong anti
microbial activity as a chemical defense against the
soil-borne microorganisms.

13.  Permealization:-
• Berberine secrets from cell culture of T. minus
used as a model system for permealization.
• Use of chemical agents (dimethyl sulphoxide,
chloroform, triton etc), ectroporation and
continuous ultrasound waves were used to
release the product is called permealization.
• This method is not good for large scale
cultures or continuous use because use of
chemical and electricity reduce the viability of
cells to a minimum or severely damage the

14. Extraction of alkaloids
Leaf/Root/Bark/Latex/Resin used as raw material
Refluxing , Soxhlet extraction , Agitation with cold solvent
with methanol or ethanol
Filtrate is obtained
Filtrate is evaporated to dryness
Residue is mixed with 1% sulphuric acid

15. Chloroform is used to separate aqueous phase using separating funnel
Non aqueous phase contains :-
Non- alkaloid compounds
Aqueous phase contains:-
Alkaloid compounds
pH change to 9 – 10 using
ammonium solution

16. Chloroform is used to separate aqueous phase using separating funnel
Non aqueous phase contains :-
Alkaloid compounds
Aqueous phase contains:-
Non -alkaloid compounds
Evaporation of organic phase
TLC, GLC, HPLC techniques are
used for purification

17. New Discoveries
 (Figueiredo et al., 2010):-
Alkaloids were isolated from the roots, bark and
leaves of Tabernaemontana salzmannii
(Apocynaceae).

18.  (Chen et al., 2011):-
Isolate alkaloids from Kopsia hainanensis, a
native medicinal plant from Hainan, China.

19.  (Kuo et al., 2011):-
Isolate alkaloids from Lobelia chinensis.

20. Conclusion
• Technology of alkaloid production has been
developed on the basis of experience.
• This technology has industrial potential but still
requires improvement in product yield.
• Identification of genes has just began and much
more work is required before generalization of
methodology can be achieved.
• Improvement in yield and technology of
bioreactors will be helpful in production of

21. Prospects
• Alkaloid production in plant cell ant tissue culture has
come to a point where a breakthrough is required to
apply this technology at an industrial level.
• Industry is still not ready to accept these capital
intensive methods for the production of secondary
metabolites.
• With the increased demand in coming decades for plant
based natural products such as anticancer, anti HIV,
food colorants, flavor, dyes health supplements etc:-
• Interest has revived in the production of natural