2. Fungal Secondary MetabolitesBY
M.S.SELMAN

3. Before I came to China I was always watching phrase (made in China) to almost anything in the markets

5. So within my mind I thought that even women are Made in China

6. So in fact, I expected to see only these scenes
So in fact, I expected to see only these scenes .... factories and industry in just about every place and only urban areas and No Nature

7. But I fortunately found just the opposite where I found stunning nature and especially here in Chongqing That probably indicates the presence of a good biodiversity .

9. Biodiversity probably means that a lot of living organisms share the same environment and compete with each other and also compete with human .

10. So what are the importance of biodiversity ?

11. Biodiversity is very useful in terms of food security and combat climate change and reduce global warming and maintain the stability of environmental cycles and to ensure that there was no cut in food chains .

12. We can use some of the organisms that live in the environment in this competition in the production of some effective and dynamic compounds that fail the industry for providing or which may be manufactured industrially costly .

14. Fungi on top of importance organisms to humans and has widespread in the environment

15. The Fungi
Fungi are one of the largest group of organisms , world wide distributed and play a vital role in ecosystems and as one of the most important tool in Biotechnology

16. Fungi & Biotechnology
BIOTECHNOLOGY means “ Use of Fungi to produce, develop, improve, remove or sustain of a particular thing which can be useful for man and nature “
The most important use of fungi is the production of Secondary Metabolites
Such as Antimicrobial Bioactive Substances that can be beneficial for Medical therapy

17. The economic significance of Fungi
Fermentation technology
Enzymes production technology
Production of acids and chemicals
Cultivation of fungi for protein
Food processing by fungi (Bread, cheese)
Fungi can be used in many applications in (industry, agriculture, medicine , and environment)
Production of Bioactive compounds (Antibiotics)

18. Secondary metabolites
Organic compounds , with low molecular weight ,which are not essential for fungal growth but their natural production have certain significances.

19. Why secondary metabolites are produced?
They are chemical compounds produced by many fungi growing on substrates. They play a role in competition ,antagonism and self-defense mechanisms against other living organisms to allow the fungus to occupy the niche and utilize the food

20. Production of SECONDARY METABOLITES
Compounds produced during the stationary and decline phases of the fungal growth

21. They are vary in structures and biosynthetic pathways
They are vary in structures and biosynthetic pathways . So they differ among the fungal species and isolates

22. Fungal metabolites structures

23. Fungal secondary metabolites can be obtained by growing fungi in cultures under certain conditions (Penicillin is best example)

24. Types of Fungal secondary metabolites 1. Strobilurin (antifungal) 2
Types of Fungal secondary metabolites 1. Strobilurin (antifungal) 2. Gibberellins (growth Hormons) 3. Herbicides (control weeds) 4. Mycotoxins (poisneous) 5. Insecticides ( control insects) 6. Enzymes (proteins) 7. Pigments (dyes) 8. Antibiotics (drugs) 9. Pharmacological drugs gs

25. Antibiotics
Antibiotics: Are chemical substances produced by fungi which has the capacity to inhibit the growth of / and or even kill other microorganisms . The action of an antibiotic is a selective in nature .

26. History of ANTIBIOTICS discovery
(1928) Alexander Flimmings discovered the Penicillin
(1935) Prontosil, the first sulfa drug was discovered by Gerhard Domagk
(1943) Andrew Moyer, industrial production of Penicillin
(1943) Selman Waksman discovered the Streptomycin from soil Bacteria

27. (1955) Tetracycline was patented by Lioyd Conover
(1957) Nystatin was patented
(1981) Smith Kline Beckham patented Amoxillin
Since then many antibiotics have been found and still continued to discover more…….

28. Antibiotic can be divided according their bioactivity into:
Antifungal antibiotics
Antibacterial antibiotics
Antiviral antibiotics
Antitumoral antibiotics

29. Mechanism of antibiotic action

30. Steps to such studies 1. Sample collection and fungal isolation:
Sample are collecting from different sources (soil , water , air , rhizospher , rhizoplane ) for fungal isolation
Isolation, purification and identification of fungi on solid media
Screening of the inhibition bioactivity of each fungal species against the selected pathogenic microorganisms including:
A. Bacteria B. Candida C. Dermatophytes D. Other fungi
by using fungal discs on solid media.
2. Fungal cultures Extraction:
The selected fungal isolates will be grown in specific liquid culture
(using batch large bottles) under shaking condition for two week incubation period at 25 C

31. 3. Antimicrobial bioactivity assay
Filter paper discs (0.6 mm) after being sterilized by autoclave are socked in each fungal crude extract solution for 5 min.,
Filter paper discs with extracts are placed on the surface of agar medium in Petri-dishes streaked with 0.2 ml of bacterial suspension of Escherichia coli and Staphylococcus aureus, Candida albicans, Microsporum gypseum, Fusarium oxysporum.
Plates are incubated at 37 ◦C for 24 hr, an appearance of inhibition zones around the filter paper disc indicating the bioactivity of crude metabolites of the tested fungal isolates (Casals, 1979).

33. 4. Extraction of the fungal filtrates
Five discs (0.5 mm diam.) are cut from the axenic fungal culture of each isolate by using a cork borer and amended into PD liquid medium in 500 ml flasks (with triplicates) and incubation at 25 ◦C for 2 weeks on a rotary shaker.
Fungal cultures are filtered on Whatman No 1 filter paper and the pH adjusted at 3 for each fungal filtrate.
Filtrate is extracted in ethyl acetate (1:1 vol) by using separating funnel. The organic layer is collected by dehydration of water by using Na2SO4. The filtrate is filtered again and place in Petri dishes then leave to be dried at room temperature.
100 ug of the dried extract is dissolved in 1 ml ethanol as stock extract solution to be used for further experiments.

34. This is separating funnel

35. 5. Minimal inhibitory concentration test
The MIC values will be determined by the standard serial dilution assay (McGinnis, 198o).
Different concentrations of the crude fungal extract are used. Filter paper discs (0.5 mm) are soaked in each extract concentration and will be tested against each selected microorganisms on solid medium. The appearance of inhibition zone around the disc indicates an inhibitory action.

36. 8. Purification of fungal extracts
6.Cytotoxicity test
Cytotoxicity of the fungal crud extracts was examined by using human RBC
following a previously described method ( Xian- guo and Ursula, 1994).
7. Chemical analysis of fungal crude extracts
Fungal culture extracts will be chemically analyzed for alkaloids, phenols, amino acids, flavenoides and Tannins according to following the described method
8. Purification of fungal extracts
Crud extract will purified by using TLC and HPLC technique.
9.Identification and characterization of the purified extract
By using GC –Mass and HNMR technique

37. After finding out the optimal conditions for the growth of fungus can be used the continuous cultures or fermenter advice for the purpose of increasing production.

38. Or we can increase the production of fungi secondary metabolits through the introduction of genetic mutations and genetic engineering techniques