The Presentation Deals with the Usage of few Edible Basidiomycetes Mushroom Fungi in Free Radical Scavenging

1. Edible Mushroom Fungi as
Antioxidants
P. Balaji
Head
PG and Research Centre in Biotechnology
MGR College,
Dr. MGR Nagar, Hosur, Tamilnadu, India

2. Nature alone is antique
and
The oldest art, a mushroom
- Carlyle

3. Mushrooms
 Mushrooms are large macroscopic fruiting bodies of either Ascomycetes or
Basidiomycetes fungi.
 Basidiomycetes, one of the four branches of monophyletic group of Eumycota (True
fungi), account for about 35% of the fungal species currently described and is of both
ecological and industrial importance.
 There are approximately 1,50,000 mushroom genera among which only 10% are
identified.
In India, mainly three kinds of mushrooms are cultivated,
1. White button mushroom (Agaricus bisporus), accounts for more than 90% of the total
production.
2. Oyster mushroom (Pleurotus sp.) is grown mostly in central and Southern India.
3. Tropical or Paddy straw mushroom (Volvariella volvacea) is grown in small scale in
Kerala.

4. MyceliumMycelium (gathering of
individual Hyphae)
Stipe
Annulus
Pileus (or Cap)
Gills (or
Lamellae)
Lower veil
Upper Veil
Hymenium
(interior Spore-producing layer)

5. Why Mushrooms?

6. International Journal of Medicinal Mushrooms
Medicinal Properties of Substances Occurring in Higher Basidiomycetes
Mushrooms: Current Perspectives (Review)
Solomon P. Wasser
Department of Evolutionary and Environmental Biology, Faculty of Natural Science,
University of Haifa, Mount Carmel, Haifa, Israel; Institute of Evolution, University of
Haifa, Mount Carmel, Haifa, Israel
Alexander L. Weis
international Myko Biologies, Inc., Texas Research Park, 14785 Omicron Dr., Suite
104, San Antonio, TX, 78245, U.S.A.
pages 31-62

8. Nutritional aspects of mushrooms
Mushrooms are
 Rich in protein
 Contains high molecular weight polysaccharides
 Antioxidants
 Low in fat and simple carbohydrates
 Good source of B complex, vitamins, potassium, copper and selenium
 Lack cholesterol and vitamins A and C
 Best sources of ergosterols, triterpenoids, glycoproteins, natural
antibiotics, enzymes and enzyme inhibitors.
Food as medicine concept
 Mushrooms are the ideal candidate for fitting in the concept of “food as
medicine”.
 Edible mushrooms and different mushroom derived compounds were shown to
have prophylactic or immune enhancing effect.
 Depending on the form in which the mushrooms or their products exert their
medicinal effects, three classes of substances are defined and named as
nutraceuticals, nutriceuticals and pharmaceuticals (Chang and Buswell, 1999).

9. World production of cultivated edible and
medicinal mushrooms
Mushrooms Production
(%)
Agaricus bisporus 31.8
Lentinus edodes 25.4
Pleurotus spp 14.2
Ganoderma spp 8.4
Auricularia spp 7.9
Volvariella volvacea 3.0
Flammulina spp 4.6
Tramella spp 2.1
Hypsizygus marmoreus 1.2
Pholioto nameko 0.9
Grifola frondosa 0.5
Hericium erinaceum 0.5
Others 0.5

10. Pleurotus citrinopileatus Pleurotus eous Pleurotus cystidiosis
Pleurotus ostreatus Pleurotus eryngii Pleuoruts flabellatus
Pleurotus florida Pleurotus pulmonarius Schizophyllum commune

11. Medicinal Effects of Mushroom
Immunomodulatoty Antitumour Antimicrobial Antioxidant
(Free Radical scavenging)
Mushroom
Cell wall components
(intracellular polysaccharides) Cell wall bound (Exo polysaccharides) Extracellular
enzymes
Dietary fibre
Chitin Glucan
α Glucan β Glucan Heterosaccharide chains of
Xylose, Mannose, galactose/
Uronic acid/Proteoglycans
Glycogen
Mono/ disaccharides
Sugar alcohols

12. Bioactive compounds
Nutritional group (key structural and metabolic
compounds) which includes Proteins,
Carbohydrates, Fat, Minerals and Vitamins etc.
Non-nutritional biochemicals (secondary
compounds) which includes Alkaloids,
Flavanoids, Terpenes, Coumarins, Lectins and etc.
Mushrooms have several bioactive compounds which can be
placed under two groups of constituents by Benjamin (1995) viz.,

13. What are Mushroom Nutriceuticals?
Refined / partially refined mushroom preparations
derived from fruit bodies, fungal mycelium or the
spent culture fluid following mycelium growth in
submerged culture that possess nutritional and/or
health-promoting properties and which are consumed
in the form of capsules or tablets as a dietary
supplement

14. Market value of medicinal mushrooms
1991 -
US$ 1.2
billion
1994 -
US$ 3.6
billion
1999 -
US$ 6.0
billion
2014 -
US$ 18
billion
2019 - US$
50.03 billion

15. Map of edible mushroom species that are commonly
grown commercially all over the world

16. China is the major producer and consumer of
medicinal mushrooms
In 2017, >375 research units/institutes were
engaged on R & D of medicinal mushrooms
Some 40-50 varieties of mushroom products for use
as nutriceuticals/herbal medicines were
manufactured in more than 200 factories
About 700 mushroom-based health food products,
including >90 brands of G. lucidum products, were
registered and marketed

17. Why not in
INDIA???

18. Mushrooms in Indian History
Amanita muscaria –
Soma, the intoxicating drink of Vedic period @. 8000 B. C.
Classification of edible, non-edible, medicinal mushrooms
in Ancient medical - Charaka Sumhita, @. 3000 B.C.

19. AYURVEDIC MEDICINE
Phellinus linteus – Bhendiomba - for purifying blood in
skin diseases
Daedaleposis flavida – Snuff fungus - for reducing
bilirubin and biliviridin for jaundice
Phellinus gilvus – Phansomba – Conks used against
kidney disorders

20. Mushrooms in the history of Tamil Nadu
Podaxis pistillaris – edible gasteromycetes – Edible
gasteromycetes in traditional Tamil Medicine –Collection at
Tharangapadi – Tamil Nadu - Identified and described by
Linnaeus in the 18th Century
Lycoperdon sp – medicinal mushroom used in Shiddha
system of medicine in Tamil Nadu

21.  ~77% of all medicinal mushroom products were derived from
fruiting bodies, which have either been cultivated commercially or
collected from the wild
 Only ~21% of all products were derived from fungal mycelia
 Approximately 2% were derived from culture filtrates
Feedstocks for mushroom products

22. Mycelial and culture broth-based
products increasing in importance
• Demands for increased safety and quality control, and for year-
round production
• Submerged culture processes can readily be standardized under
controlled conditions
• Downstream processing of active components released into the
culture medium usually involve relatively simple procedures

23. Production methods
Powdered whole
wild/cultivated fruit
bodies and mycelium
Hot water / EtOH /
MeOH of extracts
of wild/cultivated
fruit bodies or
mycelium
Various
combinations of the
above

24. Application of agro-industrial
residues in bioprocesses on the one
hand provides alternative substrates
and on the other help in abating
pollution.
Vast quantities of renewable
lignocelluloses wastes are generated
every year in developing countries
like India with economies which are
basically agricultural.
Mushroom species have much
diversity due to their adaptation to
the varying climatic conditions as
well as their ability to degrade locally
available lignocellulosic substrates.
Utilization of agricultural residues in
biological process is hindered by the
recalcitrant nature of lignin or by the
presence of toxic chemicals if any in
these resources.
In recent years there has been an increasing trend towards more efficient
utilization of agro-industrial residues for the production of bulk chemicals and
value added fine products such as ethanol, single cell protein, mushrooms,
enzymes, organic acids, aminoacids, biologically active secondary metabolites etc.

25. Free radicals
 molecular species capable of independent existence and which contains
one or more unpaired electrons
 formed as byproducts of normal metabolism include superoxide anion
(O2-), hydrogen peroxide (H2O2), singlet oxygen (O2
), hypochlorus acid
(HOCl) and hydroxy radical (OH).
 All these products are collectively referred to as reactive oxygen species
(ROS), oxy radicals (or) oxygen free radicals (OFR)

26. Antioxidants
 Any substance that when present at low concentration compared to those of
oxidizable substrate, significantly delays or prevents oxidation of that
substrate.
 Variety of antioxidant defense systems are operative, including enzymic and
non-enzymic antioxidants.
 Enzymes directly involved in detoxification of reactive oxygen species (ROS)
are superoxide dismutase (SOD), catalase and glutathione peroxidase (GPx)
and small molecules such as reduced glutathione (GSH).

27. Endogenous and exogenous factors inducing ROS generation

28. Major ROS defense mechanisms in the
organism

29. Synthesis of Silver nanoparticles
 Aqueous solution (1mM) of silver nitrate was prepared using deionized water
for the synthesis of silver nanoparticles.
 5 ml of mushroom broth/ extract was added into 95 ml of aqueous solution of 1
mM silver nitrate for reduction into Ag+ ions and heated on water bath at 75°C
for 60 min.
 Reduction of silver nitrate to silver ions was confirmed by the color change
from colorless to brown.
 The formation of silver nanoparticles was also confirmed by spectrophotometric
determination.
 The fully reduced solution was centrifuged at 5000 rpm for 30 min.
 The supernatant liquid was discarded and the pellet obtained was redispersed in
deionized water.
 The centrifugation process was repeated two to three times to wash off any
absorbed substances on the surface of the silver nanoparticles.

30. Synthesis of Gold nanoparticles
 Aqueous solution (1 mM) of Chloroauric acid (H[AuCl4]) was prepared using
deionized water for the synthesis of gold nanoparticles.
 1.5 ml of mushroom broth/ extract was added into 34 ml of aqueous solution of
1 mM chloroauric acid for reduction into Au3+ ions and incubated at 29°C for
24 hrs.
 Reduction of Chloroauric acid to Au3+ ions was confirmed by the color change.
 The formation of gold nanoparticles was also confirmed by spectrophotometric
determination.
 The fully reduced solution was centrifuged at 6000 rpm for 11 min.
 The supernatant liquid was discarded and the pellet obtained was redispersed in
deionized water.
 The centrifugation process was repeated two to three times to wash off any
absorbed substances on the surface of the gold nanoparticles.

31. Assay of antioxidant activity of synthesized Gold nanoparticles and Silver
nanoparticles
Name of the
extract
Total Flavonoid
Content
Ferric-Reducing/
Antioxidant
Power
ABTS Assay Metal Chelating
Activity
Phospho
molybdenum
Assay
Assay of
Superoxide
Radical
Scavenging effect
RE (mg/ml)
Scavenging effect
Fe ( II) (mg/ml)
Scavenging effect
(mg/ml)
Scavenging effect
EDTA (mg/ml)
Scavenging effect
AAE (mg/ml)
Scavenging effect
(%)
AuNPs AgNPs AuNPs AgNPs AuNPs AgNPs AuNPs AgNPs AuNPs AgNPs AuNPs AgNPs
Pleurotus
citrinopileatus
5166.66 11.67 359.64 317.99 6925.46 14012.92 2.43 3.03 5.55 1.67 3.61 17.82
Pleurotus eous 4462.96 26.12 789.47 293.86 10043.94 13688.92 1.65 2.89 20 3.89 62.93 76.79
Pleurotus
cystidiosis
5000 17.23 574.56 322.37 10590.69 14154.67 2.97 2.73 2.22 2.24 47.52 20.49
Pleurotus
ostreatus
4388.88 16.12 15.35 269.74 7269.70 14114.17 2.65 2.62 3.88 17.78 73.89 56.19
Pleurotus
eryngii
4638.88 45.56 65.78 315.79 7897.45 13688.92 2.84 2.87 25 6.67 73.23 32.90
Pleuoruts
flabellatus
4583.33 19.45 359.64 287.28 9517.44 14053.42 2.78 2.98 28.33 8.34 53.91 52.25
Pleurotus
florida
4750 37.23 789.47 324.57 6358.46 12797.93 2.93 2.91 2.77 7.23 55.46 1.83
Pleurotus
pulmonarius
5416.67 17.78 385.96 293.86 9659.19 13871.12 3.05 2.75 23.33 5.56 76.79 27.39
Schizophyllum
commune
4527.77 2.78 385.96 293.86 11684.18 14114.17 2.38 2.73 13.33 10.56 40.44 66.64

32. Concluding Remarks
There is growing experimentally-based evidence to suggest that dietary supplements
based on bioactive compounds extracted from mushrooms (mushroom
nutriceuticals) increase resistance to disease and, in some cases, causes regression
of a diseased state
They have extraordinary low toxicity, even at high doses, and are apparently lacking in
various side effects that frequently accompany the use of synthetic drugs
Since mushrooms have a long tradition as a food source, many mushroom feedstocks
are categorised as ‘generally considered safe’
Increasing interest in mushroom nutriceuticals is likely to continue worldwide in view
of the challenges and opportunities they represent, and their economic value may
ultimately surpass that of mushrooms currently produced for food

33. Thank you