Basics of fungi for medical field

1. FUNGAL METABOLITES – AS A STORE HOUSE OF
BIOACTIVE NATURAL PRODUCTS
Dr. V. RAMESH
Assistant Professor
Department of Botany
Vivekananda College
Tiruvedakam West – 625 234
ramesh.vnr09@gmail.com
+91 9842629245

2. What is fungi?
Mycology – The study of fungi
Micheli – Father of mycology

3. KINGDOM FUNGI
To date 400,000 species of fungi have Been
discovered.
400,000 species, most are saprophytic
 (0.5%) pathogens of humans/animals
(8%) plant pathogens
Over 65% of plant diseases are caused
by fungi

4. Kingdom Fungi
Fungi are eukaryotic
They have a nuclei & mitochondria
They are Heterotrophs
They depend on other organisms for food
They may be unicellular or multicellular
Main Characteristics of Fungi

5. Linnaeus – Plants.
Whitaker (1969) separate kingdom.
.
Different thoughts of Fungi

6. Fungi lack chlorophyll
Cannot produce their own food
Most are saprophytes
Some are parasites
Most fungi have cell walls made of chitin…
Reasons Fungi Are Different From Plants

7. Air Water Living leaf/stem surface
Interior of dying plant biomassSoilInterior tissues of living plants
Where Can we see Fungi?

8. Mammals/Birds/Other
animals
Insects
Roots/the rhizosphereHuman beings
Litter of all kindsFungi

9. Plant Body of Fungi
Mycelium – a mass of connected hyphae
Usually hidden in the soil, in wood, or
another food source
Hyphae - network of thin thread-like structures

10. 1. Zygomycota - Bread Molds
Rhizopus – black bread mold
2. Oomycota - Water Molds
Water mold, potato blight,
Mildew
3. Ascomycota - Sac Fungi
Yeast, morels, truffles
4. Basidiomycota - Club Fungi
Mushrooms, puffballs, bracket
fungi, rusts, smuts, toadstools
5. Deuteromycota - the Fungi Imperfecti
Major groups of Fungi

11. REPRODUCTION
• Most fungi reproduce Asexually and
Sexually by spores
• ASEXUAL reproduction is most common
method & produces genetically identical
organisms
• Fungi reproduce SEXUALLY when
conditions are poor & nutrients scarce

12. SEXUAL REPRODUCTION
• Haploid 1n hyphae from 2 mating types (+ and -)
FUSE (Fertilization)
• Forms a hyphae with 2 nuclei that becomes a
ZYGOTE
• The zygote divides to make a SPORE

13. ASEXUAL REPRODUCTION
• Fragmentation – part of the mycelium
becomes separated & begins a life of its
own
• Budding – a small cell forms & gets
pinched off as it grows to full size
– Used by yeasts

14. SPORES
• Spores are an adaptation to life on land
• Ensure that the species will disperse to new
locations
• Each spore contains a reproductive cell that
forms a new organism
• Non motile
• Dispersed by wind

15. Reproduce by spores
• Spores may be Formed:
– Directly on hyphae
– Inside sporangia
– On Fruiting bodies
Amanita fruiting body
Pilobolus sporangia
Penicillium
hyphae

16. ECONOMIC SIGNIFICANCE OF FUNGI
Industry – Proteins, Enzymes, alcohol & Organic
acids
Agriculture –Biocontrol agents & Growth Hormones
Food for Human Beings
Environment – Decomposers
Medicine – Antibiotics & Anticancer agents

17. Natural Products

18. Bioactive metabolites
Primary Metabolites
Substances that are widely
distributed in nature,
occurring in all organisms.
Seeds and vegetable storage
organs.
Needed for general growth
and development.
e.g. Amino acids, starch,
sugars, vegetable oils, etc.
Secondary Metabolites
Biosynthetically derived
from primary metabolites.
Limited in distribution being
found usually in specific
families.
Not necessary for growth
and development.
May serve as pollination
attractants, environmental
adaptations, or protection.
e.g. Alkaloids, Terpenoids,
Steroids, Flavonoids etc.

19. Fungal Secondary Metabolites
Herbicides (Control weeds) - Phytophthora palmivora
C. gloeosporioides
Mycotoxins (Poisonous) - Amanitin
Insecticides (Control insects) – Beauveria bassiana
Enzymes (Proteins) – Amylase, Protease, Invertase
Pigments (Dyes) Hydnellum peckii – Beige colour
Pharmaceutical drugs
Antibiotics drugs and antitumor agents

20. Antibiotics
Chemical substances produced by living organisms which has
the capacity to inhibit the growth of / and or even kill
other microorganisms.
Literal translation
• anti – against
• biotic – living things

21. Action of antibiotics

22. Since then many antibiotics have been found and still
continued to discover more…….
Overuse of antibiotics can damage our normal/good
bacteria.
– Many antibiotics prescribed by
the doctor are broad
spectrum
– These kill the body’s good
bacteria as well as the bad
– With the good bacteria gone
there is more room for bad
microbes to invade!
Antibiotic Resistance

23. Destruction of normal flora
allows pathogenic pathogens to
dominate

24. 25/03/16

25. MECHANISMS FOR
ACQUIRING RESISTANCE

26. How antibiotic resistance
can be prevented
– Antibiotics should be the last line of defence NOT
the first
• Most common infections will get better by
themselves through time, bed rest, liquid intake
and healthy living.
– Only take antibiotics prescribed by a doctor
– If prescribed antibiotics, finish the course.
– Do not use other peoples or leftover antibiotics
• they be specific for some other infection

27. ANTITUMOR AGENTS
Cancer is a group of diseases that are characterized by the loss of
control of the growth, division, and spread of a group of cells leading to
a primary tumor that invades and destroys adjacent tissues
Chemical substance which is kill the cancer cell
Vincristine
Paclitaxel
Doxorubicin
What is Cancer?

28. Types of cancer
Based on function/locations of the cells
Carcinoma a tumor derived from epithelial cells, those cells that line
The surface of our skin and organs (80-90%)
Sarcoma – derived from muscle, bone, cartilage, fat or connective
tissues
Leukemia - derived from WBC
Lymphoma - a cancer of bone marrow derived cells that affects the
lymphatic tissues
Myelomas – a cancer involving the WBC responsible for he production
of antibodies

29. Common Treatments
 Surgery
 Direct removal of tumor
 Radiotherapy
 Using ionizing radiation to control
malignant cells
 Chemotherapy (Antitumor Agents)
 Using chemicals to kill actively
dividing cells

30. Mechanism
of
antitumor
agents

31. Natural products from Micro fungi
1928 - Alexander Fleming discovered
the Penicillin
Effective against
Bacterial infections
Inhibit the cell wall
synthesis

32. Cephalosporin C
Acremonium chrysogenum
first natural product of a
marine-derived fungus (1946)
Sorbicillactone A
Penicillium chrysogenum
from sponges belonging to
the genus Ircinia (2003)
Conti….

33. Paclitaxel (Taxol)
originally isolated from the Pacific yew tree, Taxus brevifolia, but later
also reported from endophytic fungi, including Taxomyces andreanae and
Pestalotiopsis microspora (1993)
Conti…….

34. Conti…….
Fusarium oxysporum
F. oxysporum
P. microspora

35. Conti…….
Aspergillus sp.
Polycladium inflatum
Pencillium brevicompactum

36. Cytotoxic metabolites Alternaria sp. from the Egyptian plant
Polygonum senegalense
Conti…….

37. Bioactive compounds from
macro fungi
Krestin - Trametes versicolor – Anticancer activity
Lentinan - Lentinus edodes Anticancer activity
Schizophyllan - Schizophyllum commune Anticancer activity
Cytochalsins, Xylaramides, Xanthones Griesofulvins – Xylaria sp. -
antifungal and anticancer activity
Ganodermin - Ganoderma lucidum Antifungal Activity (Wang and Ng,
2006)
Agrocybin - Agrocybe chaxingu - Antifungal Activity

38. Cont……

39. Isolation of Bioactive Metabolites From Fungi
Leaf
Sterilization
70 % ethanol For 1 Min
0.5% sodium
hypochlorite for
30 s
Washed water
for 3 min
Dried with
sterilized filter paper
Inoculation
Isolation of endophytic fungi
Growth of Fungi

40. Colony morphology of Hypomyces sp. Conidiospore
Colony morphology of Alternaria sp. Conidiospore
Endophytic fungi isolated from the
Medicinal Plant

41. Colony morphology of Pestalotiopsis sp.
Colony morphology of Colletotrichum sp.
Conidiospore
Conidiospore
Conti…….

42. Macro fungi or Mushrooms
c
Xylaria sp.

43. Fungal culture extraction
The selected fungal isolates will
be grown in specific liquid
Medium
Incubation 21 days
Collection of crude extract
Extraction of culture filtrates
with organic solvents

44. Crude extract
Preliminary Screening

45. Purification of Crude extract
Crude Extract Thin Layer Chromatography
Column Chromatography
Purified Compound

46. Anticancer Activity
Purified Compound
96 Well Microplate

47. Control 8 h Control 24 h
40 µg 40 µg
40 µg40 µg
Treated cells exhibited
condensed
form of nuclear
materials.
That is apoptosis
Morphological Changes

48. Structure Elucidation of Natural Products

49. THANK YOU ALL