This document discusses the production of ergot alkaloids. Ergot alkaloids are produced by fungi in the genus Claviceps that parasitize grasses. There are several methods for producing ergot alkaloids, including obtaining them from sclerotia harvested from infected rye plants, chemical synthesis from fermentatively produced lysergic acid, submerged fermentation using Claviceps species, surface culture fermentation, and semicontinuous fermentation using immobilized mycelium. Regulation of alkaloid production in culture is dependent on phosphate levels in the growth media.

1. MIC 410: Advances in Microbial Technology
Unit 2: Microbial production of therapeutic agents
• Ergot alkaloids
-Dhritiksha M. Baria
(PhD research scholar)
Department of Microbiology and
biotechnology, Gujarat University,
Ahmedabad

2. ▪ Occurrence and Significance
▪ Structure
▪ Biosynthesis
▪ Production of Ergot alkaloids
1. Chemical synthesis
2. Obtaining from sclerotia
3. Production by fermentation
4. Surface culture
5. Semi-continuous transformation
▪ Regulation of alkaloid production in cultures

3. What are alkaloids ?
Alkaloid, any of a class of naturally occurring organic nitrogen-
containing bases. Alkaloids have diverse and important
physiological effects on humans and other animals. Well-known
alkaloids include morphine, strychnine, quinine, ephedrine,
and nicotine.
Alkaloids are found primarily in plants and are especially common
in certain families of flowering plants. More than 3,000 different
types of alkaloids have been identified in a total of more than
4,000 plant species.

4. The chemical structures of alkaloids are
extremely variable. Generally, an
alkaloid contains at least one nitrogen
atom in an amine-type structure—i.e.,
one derived from ammonia by
replacing hydrogen atoms with
hydrogen-carbon groups
called hydrocarbons. This or another
nitrogen atom can be active as
a base in acid-base reactions. The name
alkaloid (“alkali-like”) was originally
applied to the substances because, like
the inorganic alkalis, they react with
acids to form salts.
Structure of alkaloids:

5. • The function of alkaloids in plants is not yet understood. It has
been suggested that they are simply waste products of plants’
metabolic processes, but evidence suggests that they may serve
specific biological functions.
• In some plants, the concentration of alkaloids increases just
prior to seed formation and then drops off when the seed is ripe,
suggesting that alkaloids may play a role in this process.
• Alkaloids may also protect some plants from destruction by
certain insect species.
Functions of alkaloids

6. ➢ Alkaloids, some of which have therapeutic applications, are
generally derived from plants, but the ergot alkaloids are
produced by fungi.
➢ Ergot alkaloids were first obtained from the sclerotium of the
parasitic ascomycete Claviceps purpurea, which develops on rye
and other grasses.
➢ Presently there are over 40 known ergot alkaloids produced by
various Claviceps strains.

7. Production of Ergot alkaloids
1.Chemical Synthesis:
• Lysergic acid which is produced fermentatively can be chemically
transformed into desired alkaloids.
• Ergoline derivatives used medically are produced commercially
using this method, with the exception of peptide alkaloids.
• Chemical synthesis of ergot alkaloids is possible, but currently this
is not cost effective.

8. 2. Obtaining alkaloids from sclerotia: About 200-500 kg of scler-otia
can be harvested from 1 hectare (2.47) of rye.
Rye flowers
Mechanically inoculated with conidia
Fungus grows in the ovary tissue
Forms conidia which serve as the inoculum for an extensive plant
infection

9. Disadvantage of method
Harvest is only possible once a year and the extent of infection is extremely
dependent on the weather. Since 1978, this production method has been
replaced in Switzerland with the fermentation method.
3. Production by fermentation: Three species of Claviceps are currently
used in the production of alkaloids by fermentation:
Claviceps paspali α-Hydroxyethyl lysergamide
C. Fusiformis Clavines
C.Purpurea Ergotamine, ergosine, ergocristine

10. ▪ Good alkaloid formation seems to be linked to the ability to metabolize high
sucrose and citrate concentrations, but at the same time, the medium must not
contain any phosphate.
Disadvantage of method :
▪ Scale-up has been problem since the production strains are sensitive to the
shearing action to the impellar but at the same time exhibit high oxygen
requirement
▪ Maintenance of sterility is also difficult for long fermentation periods
▪ Antifoam agents usually cause lower yields

11. tree
Homogenized mycelium
of a 5-10 day agar slant
culture
Growth in shake
flask
Preculture
Production culture
Conidium
suspension

12. 4. Surface culture:
▪ A process has been described for the large scale production of ergot
alkaloids using surface cultures of C. purpurea grown under sterile
conditions.
▪ The advantages of this process is that a higher proportion of the more
desirable ergot amine and ergotoxine alkaloids are formed.
5. Semicontinuous transformation:
▪ A process has been developed for semicontinuous alkaloid production
using immobilized mycelium of C. purpurea and C. fusiformis

13. ▪ The best results have been obtained by immobilization in 4% calcium
alginate gel. At concentration of alginate up to 8%, the overall yield of
alkaloids was increased by 35%.
▪ To prevent contamination problems during the long incubation period with
the immobilized mycelium (200-400 days), an antibiotic such as
chloramphenicol was added.
❖ Regulation of ergot alkaloid production in cultures
Ergot alkaloid production exhibits a typical phosphate regulation

14. Thank you