Applications of fungi

Applications of Fungi for UG and PG students for quick go through

1. APPLICATIONS OF FUNGI
BY
SOURADEEP BANERJEE

2. INTRODUCTION TO FUNGAL USES
Primary production: - concerns with the commercial products of fungi (food) and
the fungal products (soy sauce) that are consumed by the human and animal
Industrial production: - concerns with the processes in fungi which maybe
exploited in the industry (antibiotics) or products (like pest and pathogen control).
In principle, to exploit all the fields which may directly or indirectly benefit human

3. USES OF FUNGI
•As biological control
•Weed control
•Plant pathogen control
•Nematode control
•Insect control
•Mushroom production
•Disease of fungi
•Industrial production
•Fermentation
•Food processing
•Fungal drugs
•Paper industry

4. A) FUNGI AS A BIO-HERBICIDE
Plant pathology is a large field to study and we will talk about it later, but the
basics of the subject is to know about the host-pathogen interaction between
plants (majorly economical plants) and their respective pathogen (majorly fungi).
Generally pathogen destroys the normal life cycle of economical plants, which are
of great interest for us.
Not only economical plants are their target, their hosts maybe any other plants
too
So how about if we use these fungi as pathogen to control weeds (which also
harms the normal life cycle of the crop plants)
So when biological pests are used to control growth of weeds it is known as bio-
herbicide

5. BIO-HERBICIDE IS USED TO CONTROL….
 to control the rate of growth or fecundity of weedy hosts
 to control the vigour of the weeds so that they loose in competition to crop
plants
Also used as poison
In perennial weeds, even longevity is reduced
The growth of weed is reduce to such an extent that they are no longer
harmful economically, but……..
The pathogen will not eliminate the weed completely as that would eliminate
the pathogen too, so there is a chance of flourishment of weeds again

6. THE “CLASSICAL STRATEGY”
Attempts to control the weeds was by isolating the weeds and pathogen both into
a separate environment, and let the pathogen do it’s work. If successful this
causes the weeds to grow less and the pathogen to flourish upon them.
A very few cases are documented which are successful.
Main type of fungi successful in these method is- ‘Rusts’, for the following
reasons:
Air-borne dissemination
Adaptable to wide variety of environment
Can cause epidemics in ideal condition
Very specific for their hosts

7. EXAMPLES OF CLASSICAL CONTROL METHOD
•In Australia, Puccinia chondrillina has been used to control a wheat weed’s
growth to a tolerable number, called Chondrilla juncea (skeleton weed)

8. •In Australia, rusts has been tested successfully to control growth of
blackberries, parthenium, rubber vine (Cryptostegia) and Mimosa pigra
•Lantana camara is a native of C.America and has a huge weedy infestations
worldwide. In Australia it was also a problem, so they released a series of
bio-controlling agents. About 25 insects was released to control their
growth, but failed due to plant’s own protective responses.
•Propodium tuberculatum, a rust fungi, was applied in 2001 for controlling the
growth of red-edged and pink Lantana sp. It grows in the sub-tropical
conditions with life cycle of 3 weeks
•Even Puccinia lantanae has also been used in laboratory but not yet released
as a biological control

9. Puccinia
lantanae
disease on
Lantana
camara
Propodium
disease in
Lantana sp

10. REQUIREMENTS FOR A FUNGI TO WORK
AS BIO-HERBICIDE
Pathogen must be highly specific and host must be highly susceptible
The pathogen must be able to sustain the growth of weeds even if the
conditions favour towards weeds
Pathogen must be able to mutate in order to attack other strains of the weed
Other controlling measures should be available if there is an unexpected
situation
Reproduction in artificial conditions

11. LIMITATIONS OF MYCOHERBICIDE
Biological
Resistivity of host
Genetic variation in hosts
Interactions with other organism on same plant
Hosts range
Genetic stability of fungus
Commercial: small return
Environmental
Germination in time
Temperature
moisture

12. FUNGI AS “FOOD”
The great example of fungi as a “food” is – Mushrooms
Long range of edible fungi are cultured throughout the world
Mushrooms or toadstool (commonly called but not literally, it has nothing to
do with toad nor stool) are spore bearing fruiting body, produced by
intermingling and anastomosed hyphal branches
Found in fungi classes basidiomycetes and ascomycetes
Generally have a stem/stipe, cap/pileus and gills underneath the pileus

13. NUTRITIONAL VALUE OF MUSHROOM
In normal cases colourful fruits provide variety of nutrients and dull coloured food
products provide less nutrient. Mushroom is quite contrary
Increased consumption of whole unprocessed mushrooms helps to control
diabetes, heart disease and obesity
They also promote a healthy complexion and hair and increased energy and lower
weight
One cup of chopped raw white mushroom contains:
15 cal
0 fat
2.2 gm proteins
2.3 gm carbs (0.7 gm fiber+1.4 gm sugar)
Rich in Vit B (B1, B2, B3, B5, B9) which helps the body to get energy from food
and help form the RBC

14. •For vegans the dietary source of Vit D is only mushrooms, as they do not
consume milk
•Pregnant mothers are often advised to take folic acid or folate or Vit B9 as
dietary supplement
•Minerals like- Se, K, Cu, Fe, P all available in fungi
•B-glucans are present in cell wall of many fungi which helps in decreasing blood
cholesterol and might influence insulin resistance boosting immunity and
decreasing obesity
•Mushrooms also have high amount of Choline in them, which helps in nerve
stimulation transfer, structure of membrane, chronic inflammation

15. USE OF MUSHROOMS FOR MEDICAL
PATIENTS
•For Cancer patients:
•high antioxidants, will help in get free of oxidative radicles formed in cells
•Se present helps in liver detoxification, tumor suppression, inflammation
•Vit D helps in cell cycle regulation
•Folate or vit B9 helps in DNA damage and repair
•For diabetes patients:
•High fiber diets will help to lower blood glucose level in type 1 patients
•Type 2 patients may have improved blood sugar, lipids and insulin level
•For heart patients:
•Fiber, K and Vit C work for better cardiovascular health. Na-K ion channel helps in
regulating blood pressure. B-glucans can reduce the blood cholesterol by 5 %.
•For immune sensitive people:
•Se is thought to increase immunity in individuals, as they help in production of cytotoxic
T cells

16. CULTIVATED MUSHROOMS
•Cultivated mushrooms such as –
•Agaricus bisporus
•Shii-take (Lentinus edodus)
•Straw fungi (Flammulina velutipes)
•Oyster mushroom (Pleurotus ostreatus)
•Chinese black mushroom (Auricularia polytricha)
•These are cultivated throughout the western world and most Asian countries
•Many are eaten fresh but maximum are dried and preserved and marketed
as canned truffles

17. COMMON OR BUTTON MUSHROOM
•Agaricus bisporous:
•Grown commercially in straw is mixed
with poultry litter and then composted
•Spawn is then added in the compost
when gets cooled
•Compost is then laid out beneath shade
at 24 deg C
•Alkaline material is added as a casing
over the surface for hyphae to grow
•Once mycelia reach surface of the
beds temperature is reduced for
fruiting, which is seen within the next 3
weeks

18. SHII-TAKE LOG CULTURE
•Lentinus edodus
•Highly prized mushroom in Japan for it’s diverse flavours and odours
•Range depends on the genotype of spawn, the selection of log incubated for
growth
•Grown mainly on log of deciduous plants which are inoculated with spore
suspension
•After extensive colonisation growth takes place when logs are kept standing
and temp drops
•Fruiting takes place in spring and continues for some years
•Cropping takes place in open and in traditional way in Japan.
•They are dried, packaged and sold throughout the year
•Auricularia and Pleurotus sp also grow on timber logs and are used for high
range of culture.

19. Shii-take fungi
Auricularia Pleurotus

20. STRAW MUSROOMS
•Flammulina velutipes
•Traditionally grown on rice straw
•Bundles are drenched and stood upright
•Then spawn is added in the straw
•Each bundles can produce fresh fundi for
several weeks
•Bundles are often mixed with cotton
(largely used) or organic wastes
•This often leads to more production
probably due to high N:C content

21. ECTOMYCORRHIZAL FUNGI
•Black truffle (Tuber melanosporum) most highly priced among these due to it’s
high flavour
•Other truffle such as Tirmania, Terfezia are also eaten
•Morchella and Cantharellus sp are also used in diet
•Truffles are harvested in Europe for many centuries specifically in France,
gradually spread to Spain, Italy and even China
•Culturing them is indirect
•Trees that support these fungi growth like oak and hezel tree, are used to produce
the maximum production
•Soil must be alkaline, calcareous, well drained
•Sapling are planted and pruned to encourage lateral growth.
•Growth seen after 5-10 years and continues for about 30 years
•Inoculated plants re mostly imported from France and gron in New Zealand and
Australia

22. Cantherellus
Black truffle
Canned truffle
Morchella

23. HEALTH RISKS FOR CONSUMING
MUSHROOMS
•Wild mushrooms have been part of the human diet for several centuries.
•Uncultivated wild mushrooms may possess threat to normal human life and even
death, as they will be rich in high amount of heavy metals and toxic substances
•Since b-glucans boost the immunity it may possess threat for the autoimmune
patients like- rheumatoid arthritis, lupus, asthma and multiple sclerosis
•Amanita phalloidis (death cap) produces -amnitin, which inhibits the normal
α
function of RNA-polymerase II

24. B) FUNGI USED TO CONTROL PLANT PATHOGEN
So first of all answer this question:- maximum of the plant diseases are caused by
the fungi, then how can fungi act as plant pathogen controller?
By interaction between fungi and fungi, that is by competition
 Interactions between fungi can be manipulated to reduce the damage caused by
one fungus on the plant. Essentially, these interactions include:
Direct hyphal parasitism
Anatagonistic reactions
Competition for resources
We will talk about each of these in details in further slides

25. DIRECT HYPHAL PARASITISM
•Direct hyphal parasitism or Mycoparasitism- one fungi derives its nutrition from
another without any benefit
•Parasite interactions maybe – Biotropic and Necrotropic
•Biotropic are those who are very specific to their hosts and extremely difficult to
manipulate
•Necrotropic on the other hand tends to kill those cells from which they derive
nutrients. Have broad host range and extremely useful for antibiotics extraction
•So which of these fungi should be used as biocontrol agents?
•Biotropic parasites right !! Because they are specific for their hosts

26. EXAMPLES OF NECROTROPIC FUNGI
•Trichoderma can act as a parasite for so many soil fungi:
•Rhizoctonia, Scerotinia, Fusarium, Verticillium
•Straminipiles like Phytophthora, Pythium
•Trichoderma grows from the host where it is an endophyte, releases some
hydrolytic enzymes that digest the wall of host prior penetration. It also
releases some toxins to reduce any responses from the host to invasion

27. C) FUNGI FOR DRUG PRODUCTION
•Extraordinary contribution of fungi on drug development for human
•Before the discovery of antibiotics or specifically penicillin, Egyptians used to
apply moldy bread on cut and infected wounds.
•It was 1928 when the first true antibiotic was developed by Alexander
Fleming, professor of bacteriology in St. Mary’s hospital in London.
•Still US played a huge role in marketing and large scale development of this
antibiotics by which huge number of WWII army and patients were treated.
•Penicillium juice can inhibit the growth of a large number of harmful bacteria
like- Streptococcus, Meningococcus, Diptheria bacillus etc.
•Fleming published his study in the british journal of experimental pathology in
June 1929
•It was the contribution of Howard florey, Ernst chain and their colleagues first
made the unstable penicillin into a stable life saving drug around 10 years
later it’s discovery
•They turned the oxford laboratory into penicillin factory

28. •For about 10-20 years fungi are involved in medicinal industries
•Medicines like anti-cholesterol, cyclosporin A (immunosuppressant), new
generation Penicillin are among the top 10-20 medicines being produced
•Each of these medicines have a turnover of more than $1 billion annually
•Recent medicines approved for human use-
•Micafungin- as antifungal agent
•Mycophenolate- prevent tissue rejection
•Rosuvastatin- reduce cholesterol
•Cefditoren- antibiotic
•Want to read more about penicillin and it’s drug discovery go to
https://www.acs.org/content/acs/en/education/whatischemistry/landmarks/flemingpenicillin.html

29. ANTI-FUNGAL DRUG DISCOVERY

30. POSITIVES AND NEGATIVES OF
FUNGI USED IN DRUG INDUSTRY
•These diversified molecules is due to the diversified metabolic pathways fungi
uses within themselves to produce such diverse final products
•Each taxa of fungi produce a different final product from the same
•On the negative, a single isolates of fungi may lose their capacity to form or
release target molecules. As the target molecules maybe expressed only under
certain specific conditions
•In more recent approaches application of hydrophobins from fungi to surfaces
helps in drug delivery in much more targeted way and also helps in
biocompatibility with implants

31. ANTIBIOTICS FROM FUNGI

32. PENICILLIN
•Penicillin- Penicillium chrysogenum, first isolated to treat bacteria
•During the early days about 1ϻg/ml was the concentration of active ingredient. But
today about 700
ϻ
g/ml are isolated as active ingredients from development of strains
•Penicillin both synthetic and natural, are
among the ß-lactam group of anitibiotics
•They attack the cell-wall formation in
bacteria by binding to the PBP’s
(penicillin binding proteins) helping in
formation of the cell wall
•PBP’s- transpeptidase, caroboxypeptidase
•It discontinues the formation of the peptide
bond b/w the L-Lys and D-Ala of tetrapeptide
chain

33. •ß-lactam group of antibiotics are very safe to use in eukaryotes as they don’t
have cell walls in them
•Still bacteria secreting ß-lactamase are resistant to this antibiotics like:
•gram positive bacteria (Staphylococcus aureus) are resistant to this antibiotic, as
they have constitutive plasmid and chromosomal production of this enzyme
•gram negative bacteria like- Pseudomonas aeruginosa, have inducible chromosomal
ß-lactamase gene in them
•Disadvantages of naturally occurring penicillin:
•Can’t be ingested orally, as it gets degraded in acidic stomach
•Sensitive to ß-lactamase
•Only work on gram positive bacteria
•Though development in medicinal sciences has discovered oral ingestions of this
antibiotics but rise of MDR’s is a serious concern in having antibiotics

34. Penicillium notatum is a
mold that frequently
grows on fruit and is the
source of the
antibiotic penicillin.

36. •Cephalosporin- another ß-lactam antibiotic molecule. Developed from
Cephalosporium sp.
•Also sensitive to ß-lactamase enzyme, thus not applicable for all those secreting
this enzyme
cephalosporin

41. GreseoFUlVin
•A broad spectrum antibiotics (antifungal agent) is Griseofulvin produced first from
Penicillium griseofulvin.
•It is a fungistatic not fungicidal
•Used to treat dermatophytes in hair skin etc.

42. sTroBilUrins
•Strobilurins excreted from fungi as soon they are produced
•It targets the ubihydroquinone oxidation centre in mammals.
•Specifically basidiomycetes of tropical region produce this diverse compounds
in huge amount
•Produced from Strobilus tenacellus

43. soDarins
•Sodarins are narrow range of antifungal agents against yeast and yeast like fungi
•It targets the protein biosynthesis process, thus becomes a good antifungal agents
against many harmful fungi attacking human

44. echinocanDins/aniDUlaFUnGin
•Echinocandins target cell wall formation are cyclic peptide molecules with long
chain fatty acid
•Semisynthetic compound used in humans is pneumocandins

46. cYclosporin a
•Primary metabolite of Trichoderma polysporum and Cylindrocarpon lucidum
•Strong immunosuppressant
•Widely used after bone marrow and organ transplantation
•Cyclic peptide consisting 11 main hydrophobic a.a
•It inhibit lymphocytes in a specific way:

47. •Calcineurin is highly conserved in phylogenetically diverse organisms
•In human pathogenic fungi Cryptococcus neoformans, calcineurin is necessary
for recovery from cell cycle arrest
•Thus application of cyclosporin A will cause death of the fungi
•The only disadvantage of application of Cylcosporin A in human is that it
supresses the immune system of human too and the side effect is a huge risk
•In human, this drug is not used as fungicide

48. GlioToXins
•Gliotoxins (Fig 1) have both immunological and antibiotic effect
•Belongs to a class of molecule called- Epipolythiodioxopiperazines (A,B,C)
Fig1

49. erGoT alKaloiDs
•Claviceps purpurea causes the St. Anthonies fire, a scourge of middle east when
the flour got infected by ergot
•Ergot contain various alkaloids, with diverse functions
•Dilates blood vessel by inhibiting noradrenaline and
sclerotin of sympathetic nervous system
•They act directly on smooth muscles of uterus causing
premature abortion
•Strongest use in intoxication due to the lysergic acid
amides
•Causes ergotism: long term effect of ergot alkaloids

50. MeDicinal Uses oF erGoT
•Due to presence of wide varieties of alkaloids they are used in medicinal purposes
a lot
•Treatment of migraine
•Vasodilation causes relief from tension and pressure
•Alkaloids are now produced by C. fusiformis and C. paspalii

51. sTaTins
•Aspergillus terreus produces a secondary metabolite called Lovostatin

52. •Phoma sp produces a type of statins called- Squalestatin

53. USES OF STATINS
•Reduce or remove low density lipoprotein from blood vessels
•Lovostatin inhibits HMG CoA redustase whereas squalistatin inhibits
squalene synthase
•Blocking these enzymes actually prevents the cholesterol in bloods
•Thus indirectly helping in controlling diabetes, pressure, fat deposition in
heart
•Statins also attracts stem cells towards damaged tissues
•Only one negative impact; one of its derived compounds causes muscle lose

54. ORGANIC ACIDS PRODUCED FROM FUNGI
Introduction
•Filamentous fungi has an exceptional record in producing organic acids
•They have quite a ability to degrade lignocellulosic biomass, and can turn certain
organic products into organic acids
•For an example, Aspergillus niger can produce hundreds of grams of citric acid/ ltr
from glucose with >80% efficiency
•But production of organic acids from fungi is not even considered in the
bioprocessing aspects of fungi (but you should highly consider it)

55. •Perhaps some of you who may find this aspect of science quite interesting, may
exploit these organisms for the better production of organic acids in this new
genomics based era
•Though till now the fungi like yeasts, Saccharomyces, Rhodotorula, Pichia etc were
of great importance biotechnologically for beverages and drugs. There has not
been much report about them in organic acids.
•Exception of all these yeasts species- Yarrowia lipolytica and it’s related yeasts has
reports for producing citric acids
•Furthermore in the near future yeasts maybe used in production of lactic acids.

56. EVOLUTIONARY SIGNIFICANCE
•But why is it that only some fungi can produce huge amount of organic acid from a
provided organic compounds but other can’t? Is it a kind of adaptation?
•Ans: Well some fungi like the filamentous one’s can produce more amount of
organic acids than any other type of fungi. The reason behind that maybe the
habitat they grow like, rocks, fruits, breads etc. the organic acid production gives
them a competitive advantage over the other fungi by decreasing the ambient pH.
•Even due to the production of organic acids by them the are of great help
maintaining ecological balance by metal detoxification and weathering of rocks

58. •From a study by Liaud et al. 2014, about 26 strains of Basidiomycota and 40
strains of Ascomycota are capable of producing organic acids and ethanol from
glucose solution and turn it into an acidic one.
•From that study after 6 days of inoculation one group mainly consisting of
Aspergillus and a few ascomycota produced acids in 4g/ltr, where the other group
contained basidiomycota and ascomycota members producing mainly ethanol.
•Acids like oxalic, gluconic and formic acids were made specifically by the
basidiomycetes members whereas Aspergillus mainly produced the oxalic, citric,
gluconic and ethanol

59. according to the final pH of growth medium after 6 days
of incubation. (orange square) Ascomycota, (sky blue
square) Basidiomycota [Liaud et al. 2014]

60. Figure 2 Hierarchical
clustering of organic acids
and/or ethanol producing
strains. Concentration were
determined by HPLC-UV or RI
analysis
and expressed as a
percentage of the maximum
concentration observed for
each metabolite and
represented by a color scale
with different intensity of
blue. Concentration of
butyric, tartaric, oxalic,
malic, citric, gluconic,
succinic acids and ethanol
were used to build distance
tree. The figure was edited
using
the Multiexperiment Viewer
software [20].

61. Highest concentrations of organic acids and
ethanol obtained at day 6 of incubation, for
each compound and the corresponding
producing strains

62. Concentrations and conversion yields for the 6
best organic acid producers

63. Repartition of the strains selected for the screening in
the Ascomycota and Basidiomycotaphyla. 40 strains
of Ascomycota representing 6 families and 26 strains of Basidiomycota representing
16 families were screened.

64. Acid Production organism( s) Application
Citric acid Aspergillus niger
Yarrowia lipolytica
Food, beverage
Pharmaceuticals, chemical
Gluconic acid Aspergillus niger Food additive
Cleaning metal surfaces
and glassware
Therapeutic metal salts
Itaconic acid Aspergillus terreus
Aspergillus itaconicus
Co-polymer
Detergents
Kojic acid Aspergillus oryzae
Aspergillus flavus
Skin whitening
Precursor for food additives
Malic acid Aspergillus spp.
Saccharomyces cerevisiae
Food additive
Synthetic polymers
Fumaric acid Rhizopus oryzae Food additive
Synthetic polymers
Lactic acid Rhizopus spp. Food additive
Synthetic polymers
Gallic acid Aspergillus spp. Dyeing
Epoxysuccinic acid Aspergillus fumigatus Precursor for tartaric acid
Adapted from Ruijter et al.

66. REFERENCE
•Magnuson, J. K., & Lasure, L. L. (2004). Organic acid production by filamentous fungi. In Advances
in fungal biotechnology for industry, agriculture, and medicine (pp. 307-340). Springer, Boston, MA.
•Ruijter, G. J. G., Kubicek, C. P., & Visser, J. (2002). Production of organic acids by fungi.
In Industrial Applications (pp. 213-230). Springer Berlin Heidelberg.
•Liaud, N., Giniés, C., Navarro, D., Fabre, N., Crapart, S., Herpoël-Gimbert, I., ... & Sigoillot, J. C.
(2014). Exploring fungal biodiversity: organic acid production by 66 strains of filamentous
fungi. Fungal Biology and Biotechnology, 1(1), 1.
•Sazanova, K. V., Shchiparev, S. M., & Vlasov, D. Y. (2014). Formation of organic acids by fungi
isolated from the surface of stone monuments. Microbiology, 83(5), 516-522.
•https://www.acs.org/content/acs/en/education/whatischemistry/landmarks/flemingpenicillin.html