Int.J.Curr.Microbiol.App.

Sci (2014) 3(4): 1073-1085

ISSN: 2319-7706 Volume 3 Number 4 (2014) pp. 1073-1085

http://www.ijcmas.com

Original Research Article

Mycelial Growth and Bioactive Substance Production of
Pleurotus ostreatus in Submerged Culture

Olfat S. Barakat and M.W.Sadik*

Department of Microbiology, Faculty of Agriculture, Cairo University, Giza 12613, Egypt

*Corresponding author

ABSTRACT

The aim of the current work is to study the potential of edible mushrooms as a
mycelial biomass source use as food additives and bioactive source
(Exopolysaccharide) for antimicrobial and antioxidant activity. Also, it aims to
Keywords study the effect of various carbon sources such as glucose, maltose, sucrose, starch,
lactose and fructose on mycelial biomass and exopolysaccharide production by
Pleurotus, Pleurotus ostereatus at different fermentation periods. The optimal period for
Myelial, mycelial biomass and exopolysacchride production was 15 days. The optimal sugar
for mycelial biomass production was starch. The optimal sugar was glucose for
Exopolysacch
high efficiency of exoploysaccharide production; antimicrobial and antioxidant
aride, activities. The concentrated exopolysaccharide filtrate of Ploroteus ostreatus
Antimicrobial, showed wide antibacterial inhibition (63.15%, 59.25%, 55.31%,36.84%,17.24% for
Antioxidant. B.subtilis, B.cereus, E.coli, Staph aureus and Pseudomonas aeruginosa,
respectivelly) when glucose used as the only carbon source.. Pleurotus ostreatus
cultivated in a medium containing glucose as the only carbon source showed the
highest antioxidant activity (59.90%).The bioactive contents of the mushrooms are
promising natural antimicrobial agents that can be harnessed as potent antibacterial
and fungi toxicants.

Introduction
Pleurotus spp. (Oyster mushrooms) 2002). In developing countries like Egypt
comprises the group of edible white-rot mushroom progress is a boon in the field
fungi with important medicinal properties of food, medicine, and in generating
and biotechnological and environmental employment.
applications. It represents a major and
untapped source of potent new In recent years Basidiomycetes and other
pharmaceutical products. A wide range of higher fungi including some recognized
activities including antitumour, medicinal mushrooms have been
cardiovascular and antimicrobial are recognized. Medicinal mushrooms have
reported in mushrooms. (Cohen et al., been re-investigated as sources of novel

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antibiotics mainly as a result of increasing and P. aureovillosus were tested against

difficulty and the cost of isolating novel four species of Gram-positive bacteria,
bioactive compounds from the five species of Gram-negative bacteria and
Actinomycetes and Streptomycetes. The one species of yeast. Pleurotus species had
research possesses an idea about the a narrow antibacterial spectrum against
antibiotic activity of some of the important Gram-negative bacteria and strongly
wild mushrooms (Karwa and Rai, 2009). inhibited the growth of the Gram-positive
bacteria tested, including Bacillus subtilis,
and M. luteus (Loganathan et al., 2008).
The petroleum ether, chloroform, acetone
and water extracts of polysaccharides from
mushroom Osmoporus odoratus has been Mushrooms are rich sources of
observed that the antibacterial activity antioxidants (Barros et al.,2007). In the
against Staphylococcus aureus, last years several protocols have reported
Streptococcus pyogenes, Bacillus subtilis, to determine their antioxidant activity
E. coli and Pseudomonas aeruginosa; the based on spectrophotometric techniques
water extract alone showed antibacterial progressively, electrochemical techniques
activity against the tested organisms and have been tested and developed as
the results were comparable with that of alternative tools for the evaluation of
amphicillin rather than chloramphenicol different food extracts, expressed in terms
(Sivakumar et al., 2006). of antioxidant power due to their
quickness , simplicity and low cost
Determination of antimicrobial activity (Blasco et al 2004). On the other hand,
profile of Lycoperdon perlatum, Ascorbic acid and phenols compounds are
Cantharellus cibarius, Clavaria common antioxidants in mushrooms.
vermiculris, Ramaria formosa, Maramius Electrochemical measurement at positive
oreades and P. pulmunarius tested against potentials will then correspond to the
a panel standard pathogenic bacteria and oxidation of total phenolic and ascorbic
fungi indicated that the concentration of acid plus all the compounds with natural
bioactive components directly influence antioxidant properties and electrochemical
the antimicrobial capability of the isolates activity which are present in foods.
(Ramesh and Pattar, 2010). (Blasco et al 2004).

Quershi et al. (2010) have studied that the This work aims to discuss: Firstly
antimicrobial activity of various solvent optimizing the conditions for maximum
extracts (40 g/ml) of Ganoderma lucidum mycelial and bioactive polysaccharides
was tested against six pathogenic species production from several types of sugars
of bacteria. Acetone extract exhibited with inoculation by Pleurotus ostreatus.
maximum antibacterial activity Secondly: Studying the antimicrobial and
(31.60±0.10), while the most susceptible antioxidant properties of Pleurotus
bacterium observed was Klebsiella ostreatus on some gram positive, gram
pneumoniae. negative bacteria, some fungi and some
yeasts.
The antimicrobial effect of ethanol
extracts of Pleurotus sajorcaju, P. florida

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Materials and Methods bioactive substance using different carbon

sources i.e. glucose, lactose, fructose,
Source of Basidiomycetes maltose, sucrose, starch with concentration
20 g/l . The fermentation medium was
Throughout the current investigation, a inoculated with 5 % (v/v) of the seed
fungal strain was tested for their potential culture and then cultivated in a 250 ml
to produce bioactive substances. Pleurotus flask containing 50 ml of MCM medium.
ostreatus was obtained from the Culture The culture was then incubated at 25°C
Collection of the Laboratory of with shaking at 200 rpm for 10 and 15
Mushroom, Food Technology Institute, days (Iwan, 2009).
Agriculture Researches Center, Cairo,
Estimation of mycelium dry weight
Egypt.
exopolysaccharide
Microorganisms
Samples collected from shake flasks were
Microorganisms used for the antimicrobial centrifuged at 5000 rpm for 20 min. The
test were gram positive bacteria: such as dry weight of mycelium was measured
Bacillus subtilis, Bacillus cereus and after drying at 70°C for overnight to a
Staphylococcus aureus .Gram negative constant weight (Iwan, 2009). Yield
bacteria like: Salmonella typhi, percentage was calculated in relation to 20
Escherichia coli and pseudomonas g/l initial sugar concentration.
aeruginosa .Pathogenic fungi like Candida
albicans and and Aspergillus niger were Estimation of extracted
used for antimicrobial tests. exopolysaccharide
Microorganisms were collected from the Certain amount of supernatant was mixed
Culture Collection of the Department of with three volumes of absolute ethanol and
Agriculture Microbiology, Faculty of left for 24 hrs at 4 C . The resulting
Agriculture, Cairo University. precipitate was then separated by
Mycelial cultivation through spore centrifugation at 5000 rpm for 10 minutes
germination (Bae et al., 2000). The dry weight
exopolysaccharide was measured after
Sabouraud dextrose agar slant was heavily drying at 70 C for overnight to a constant
inoculated with spores collected from the weight (Iwan,2009). The rest amount of
gilled mushroom, Pleurotus ostreatus. crude exopolysaccharide supernatant used
Incubation was carried out at ambient in antimicrobial assay. Yield percentage
temperature for 7 days. Several sub was calculated in relation to 20 g/l initial
culturing exercises were carried out until a sugar concentration.
pure culture was obtained. The mycelium
culture thus obtained was used as Mycelial biomass and
inoculum in subsequent experiments. exopolysaccharide production from
molasses
Fermentation medium
The clarification of molasses was done by
The polysaccharide production was carried adding 3 ml concentrated sulfuric acid to 1
out on submerged fermentation culture. Kg molasses mixed with 1000ml tap
Mushroom complete medium (MCM, water. The mixture was heated in a water
Oxoid) was used for the production of the bath to boiling for 30 minutes, and then it
was stand in refrigerator overnight and

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sterilized at 121 C for 15 minutes. days and 15 days incubation period. For
Different concentrations of molasses (2%, control experiment, 50ml of fresh PDB
4%, 8%) media were inoculated with 5% (M-lab) was added to 50ml sterile distilled
(v/v) of seed culture and then cultivated in water (without the mushrooms filtrates)
a 500 ml flask containing 100ml of MCM and inoculated with each bacterium. The
medium. The culture was then incubated at OD was measured with the growth
25 C with shaking at 200 rpm for 15 days. inhibition of the microorganism as
Samples collected from shake flasks were expressed in percentage with the equations
centrifuged at 5000 rpm for 20 minutes. of:
The dry weight of mycelium was Growth inhibitions (%)= x 100
measured after drying at 70 C overnight to
a constant weight.(Iwan, 2009). Certain
amounts of supernatant was mixed with OD1= OD of microorganisms tested in
three volumes of absolute ethanol and left medium without culture broth of P.
for 24h at 4 C. The resulting precipitate ostreatus.OD2= OD of microorganisms
was then separated by centrifugation at tested in medium with mushroom filterate
5000 rpm for 10 minutes (Bae et al., (Sterilized crude exopolysaccharide
2000). The dry weight of filterate of P. ostreatus.
exopolysaccharide was measured after
drying at 70 C for overnight to a constant Antioxidant activity
weight (Iwan, 2009). The rest amount of
the supernatant used in the antimicrobial The DPPH[2,2-Diphenyl-Picrylhdrazyl]
assay. scavenging activity was measured using
spectrophotometry (Lee et al., 2002).
Antimicrobial activities DPPH scavenging effect calculated
according to the equation. DPPH
Microbial growth by optical density scavenging effect (%)= A0- AP/A0 X
method was used to evaluate the 100.Where A0 is the absorbance of the
antibacterial activities of the culture control and AP is the absorbance of
filtrates. Bacteria for testing the sample. Vitamin C (Ascorbic acid) was
antibacterial activities were grown in used as control.(Adebayo et al., 2012).This
nutrient agar (NA) (M- Lab). The experiment was done to evaluate the
antibacterial activity of the mushroom antioxidant activity of extracted
culture filtrate (Crude exopolysaccharide) exopolysaccharide (EEPS) produced in
were evaluated by adding 50ml of media containing glucose or maltose or
mushroom filtrate to 50ml of fresh potato 2% molasses as only carbon sources. Also,
dextrose broth (v/v) and then autoclaved to evaluate the antioxidant activity of
at121 C for 15 min (Imtiaj and Lee, 2007) crude exopolysaccharide (CEPS) in
. Cooled liquid medium containing medium containing glucose as only carbon
mushroom filtrate was inoculated with source.
each tested bacterium separately in 250ml
conical flasks and incubated at 35 C for Statistical analysis
10 and 15 days incubation periods. The
bacterial growth was determined by The data in triplicate for the parameters in
measuring the optical density (OD) at various experiments were subjected to
600nm (Imtiaj and Lee, 2007) after 10 ANOVA (Analysis of variance) (Silva et
al., 2009).

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Results and Discussion (5.35%)(Table.3).

Effect of carbon source on mycelia Exopolysaccharide production by
biomass production P.ostreatus using Molasses as cheap
carbon source material
Pleurotus ostreatus grow in Mushrooms
complete medium containing starch as Different concentrations of molasses as a
carbon source; produce the highest raw material (2%, 4%, and 8%) were
mycelia biomass with high significant after tested for exopolysaccharide production
10 and 15 days of incubation period (6.29 with low cost. It was observed that
and 6.82 g/l, respectively as shown in medium supplemented with 8% molasses
(Table.1) with the highest percentage of give the maximum exopolysaccharide
biomass yield (31.45 and 34.10, production (14.05%) with the highest
respectively). significant and with the highest percentage
of exopolysaccharide yield (70.25%) after
As general results showed decline in pH 15 days incubation period (Table.4).
with incubation time and with all carbon
sources applied. It was observed that Antimicrobial activity of
decreasing in pH does not affect the exopolysaccharide produced by
mycelia biomass production as Pleurotus P.ostreatus
ostreatus produce the highest mycelia
biomass production when it cultivated in a
medium containing starch as only carbon The antimicrobial activity of
source(Table.2). exopolysaccharide in the crude
exopolysaccharide of P.ostreatus is
Mycelium production by P.ostreatus expressed by the growth inhibition of the
using molasses as cheap carbon source tested microorganisms as shown in (Table.
material. 5). Growth of P.ostreatus in medium
containing glucose as carbon source
Different concentrations of molasses as a exhibited the highest growth inhibition of
raw material (2%,4%,and 8%) were tested the most tested microorganisms using
for mycelium production with low cost. It crude and diluted exopolysaccharide. The
was observed that molasses give the crude exopolysaccharide extract showed
maximum mycelium production (18.62 varying degree of inhibition on the tested
g/l) with the highest significant after 15 microorganisms. The results of optical
days incubation period with the highest density showed that the concentrated crude
percentage of biomass yield (93.40%) with exopolysaccharide filtrate of Pleurotus
reduction in pH till 4.7 (Table.2, Fig.2). ostreatus was highly significantly
effective against gram positive bacteria
Effect of carbon sources on than gram negative bacteria after 15 days
exopolysaccharide production. incubation time .Also, results of optical
density showed that concentrated crude
The highest polysaccharides production exopolysaccharide culture filtrate was
was obtained by the utilization of glucose highly significantly effective against E.
as carbon source after 15 days of coli and S. aureus after 15 days incubation
incubation period (1.07g/l) with the time.
highest significant and with the highest
percentage of exopolysaccharide yield

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The concentrated culture filtrate of for their submerged culture. In order to

Pleurotus ostreatus showed slight investigate the effects of different carbon
inhibitory effect on Pseudomonas sources on growth and extracellular
aeruginosa (Table 5). Candida albicans is polysaccharide of Ploroteus ostreatus, the
more susceptible to the concentrated fungus was cultivated in the medium
culture filtrate of the mushroom when containing various carbon sources for 10
compared to A.niger. Crude and 15 days. Initial pH was adjusted to 5.5
exopolysaccharide produced by (Table 1 and Table 2) showe the cell
P.ostreatus in a medium containing concentration (dry weight, w/v) and the
maltose shows the highest growth extracellular polysaccharides production.
inhibition against Aspergillus niger after The maximum exopolysaccharide
10 days incubation i.e.23.80 %. production was obtained from medium
containing glucose with a final mycelial
The highest percentage inhibitory effect on biomass concentration of 1.07g/l. This
bacterial growth was recorded in B.subtilis result is in agreement with the results
(63.15%) while the least was recorded reported by other investigators, who
(3.44%) on Pseudomonas aeruginosa after demonstrated that glucose is clearly a
15 days incubation time. Results showed good carbon source for exopolysaccharide
that the highest percentage inhibitory production in submerged cultures of
effect on bacterial and fungal growth was mushrooms (Xu et al. 2003). Meanwhile,
achieved when glucose used as the only this result is in disagreement with (Iwan ,
carbon source with all microorganisms 2009) who found that the highest
under study (Table.5). exopolysaccharide production of Pleurotus
Effect of antioxidant activity of crude ostreatus is obtained in a medium
exopolysaccharide of P.ostreatus containing maltose as only carbon source.
cultivated in media containing different Meanwhile 16.6% inhibition of spore
carbon sources. germination of Aspergillus niger was
observed from the medium containing
It could be noticed that crude glucose as carbon source. The maximum
exopolysaccharide produced in a medium mycelial biomass level was found in
containing glucose shows the highest medium containing Starch. The fungus
antioxidant activity 59.9% at 150 µl with grew very well in the medium containing
the highest significant (Table.6). Starch with a total biomass concentration
Although, the antioxidant activity of of 6.82 g/l, being significantly higher than
exopolysaccharide of P.ostreatus the medium containing maltose, lactose
cultivated in medium containing 2% and other sugars. Sistorm and Michilis
molasses is lower than the antioxidant (1955) explained that fructose is next best
activity of exopolysaccharide of utilized carbon by many fungi. Often
P.ostreatus cultivated in medium adaptation is required before growth
containing pure sugars by about 10%. The begins, but then growth is often as rapid as
huge amount of exopolysaccharide with glucose. It was unexpected, therefore,
produced by P.ostreatus cultivated in a the inhibition of spore germination was
medium containing 2% molasses offsets low when fructose was used (Table 1). It
that decrease in antioxidant activity. seems that glucose as a carbon source is
necessary for production of such bioactive
Many kinds of mushrooms frequently compounds. This was explained that a
require starch, glucose, sucrose and etc.,

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Table.1 Myelium production by P ostreatus using different sugar types

Type of Mycelium production (g/l) at Biomass Yield %

Sugar different incubation period
0 days 10 days 15 days 0 days 10 days 15 days
Glucose 0.70 2.68 3.15 3.5 13.4 15.75
Maltose 0.70 5.46 6.04 3.5 27.3 30.2
Lactose 0.70 3.08 3.02 3.5 15.4 15.10
Sucrose 0.70 0.98 1.07 3.5 4.9 5.35
Starch 0.70 6.29 6.82 3.5 31.45 34.10
Fructose 0.70 1.47 1.28 3.5 7.35 6.40
LSD(0.01) 0.2805
LSD carbon sources x incubation time _ (0.01)
Figure.1 Changes in pH during mycelium production using different types of sugars. pH at
zero time was 5.5 in all treatments
pH

o
days
days

Glucose Maltose Lactose Sucrose Starch Fructose

Sugar type

Table.2 Mycelium production by P.ostreatus using Molasses as carbon source.

Molasses % Mycelium production (g/l) Yield %

at different incubation
period.
0 days 15 days 0 days 15 days
2% 0.90 7.37 4.50 36.85
4% 0.90 13.00 4.50 65.00
6% 0.90 18.62 4.50 93.40
LSD(0.01) 0.4233
LSD concentrations x incubation time _ (0.01)

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Figure.2 Changes in pH during mycelium production using different

concentrations of molasses

pH

.
Table.3 Exopolysaccharide production by P. ostreatus using different sugar types.

Types of sugar Exopolysaccharide production Yield %

g/l) at different incubation
period.
10 days 15 days 10 days 15 days
Glucose 1.04 1.07 5.2 5.35
Maltose 1.02 1.04 5.1 5.20
Lactose 0.80 1.00 4.0 5.00
Sucrose 1.00 1.01 5.0 5.05
Starch 0.80 0.90 4.0 4.50
Fructose 0.50 0.70 2.50 3.50
LSD (0.05) 0.26518
LSD carbon sources x incubation time _ (0.05)
EEPS: Extracted exopolysaccharide

Table.4 Exopolysaccharide production by P.ostreatus using

different molasses concentrations
Molasses Exopolysaccharide Yield%.
production (g/l) after 15
days incubation period.
2% 4.01 20.05
4% 9.00 45.00
8% 14.05 70.25
LSD (0.01) 0.4618
LSD concentration x incubation time _ (0.01)
The amount of exopolysaccharide at the beginning of experiment= 0.01 g/L

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Table.5 Antimicrobial activity of exopolysaccharide of P.ostreatus in mushrooms complete medium containing different types of
sugar types after 10 days and 15 days of cultivation using crude and diluted exopolysaccharide.
Carbon Incubation Inhibition of microbial strains%
Source Time (Days) Dilutions E.coli B.subtilis B.cereus Pseudomonas Salmonella Staph Candida A.niger
aeruginosa aureus albicans
10 Conc 54.16 61.64 58.22 16.12 25.58 33.33 35.51 16.66
Glucose
*
Dilut 35.41 43.83 37.97 9.67 20.93 22.22 17.22 7.14
15 Conc 55.31 63.15 59.25 17.24 26.82 36.84 36.27 17.02
*
Dilut 38.29 44.73 40.74 13.79 24.39 33.33 34.31 14.89
Maltose 10 Conc 43.75 61.64 53.16 9.67 19.76 35.18 29.90 23.80
*
Dilut 25 26.02 31.64 3.22 19.76 12.96 13.08 7.14
15 Conc 44.68 61.84 58.02 10.34 20.73 36.84 30.39 25.59
*
Dilut 25.53 27.63 32.09 10.34 20.73 28.07 25.49 14.89
Lactose 10 Conc 14.58 9.58 32.91 0 2.32 14.81 10.28 4.76
*Dilut 0 6.84 18.98 0 0 9.25 7.47 0
15 Conc 14.89 10.52 33.33 3.44 2.43 15.78 10.78 6.38
*
Dilut 6.38 7.89 19.75 3.44 2.43 10.52 7.84 4.25
Sucrose 10 Conc 33.33 36.98 45.56 9.67 15.11 22.22 23.36 14.28
*
Dilut 12.50 19.17 22.78 0 8.13 12.96 9.34 7.14
15 Conc 36.17 38.15 45.67 13.79 15.85 22.80 24.50 17.02
*
Dilut 17.02 19.73 23.45 6.89 13.41 15.78 16.66 12.76
Starch 10 Conc 20.83 20.54 26.58 12.90 9.30 12.96 17.75 7.14
*
Dilut 14.58 13.69 17.72 3.22 6.97 7.40 9.34 2.38
15 Conc 21.27 25 29.62 13.79 10.97 14.03 18.62 8.51
*
Dilut 14.89 14.47 18.51 10.34 7.31 10.52 12.74 6.38
Fructose 10 Conc 6.25 9.58 12.65 6.45 5.81 7.40 6.54 7.14
*
Dilut 6.25 8.21 12.65 0 0 3.70 6.54 0
15 Conc 8.51 10.52 13.58 6.89 6.09 10.52 6.80 6.38
*
Dilut 6.38 9.21 13.58 3.44 2.43 3.50 5.84 0
*
Dilut: Diluted 1:1 .
LSD carbon sources x incubation time x Concentration x strains _ (0.05) = 0.7693

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Table.6 Antioxidant activity of crude exopolysaccharide of P.ostreatus cultivated in media

containing different carbon sources

Exopolysaccharide Antioxidant activity of Exopolysaccharide [DPPH assay scavenging

obtained from %]
different types of 25 µl 50 µl 100 µl 150 µl
sugars
Glucose EEPS 27.34% 31.51% 41.41% 53.65%
Maltose EEPS 24.22% 29.69% 38.28% 46.61%
2% Molasses EPS 21.09% 27.60% 35.94% 41.93%
Glucose CEPS 31.25% 35.68% 46.09% 59.90%
LSD (0.01) 0.9057
LSD carbon sources x concentrations _ (0.01)
EEPS: Extracted exopolysaccharide obtained from different types of sugars
CEPS: Crud exopolysaccharide obtained from medium supplemented with glucose as carbon
source.

fungal species may have the ability to Polorteus oysterous, compared to fructose
utilize a particular carbon source for and glucose (Moore,1969).
vegetative growth but may be unable to
use it for production of specialized The crude concentrated exopolysaccharide
structure (Garraway MO and Evans, 1984) filtrate of mushrooms under study showed
a wide range of antibacterial and
It is interesting to note that little difference antifungal activity (Table.5). They
was seen in the inhibition activity of the exhibited moderate to good antibacterial
extracts from the media containing activity against the bacteria pathogens
fructose and lactose. The biomass in the tested. The filtrate of Pleurotus ostreatus
growth medium containing maltose was is very effective against E. coli and
lower than that of fructose; however the S.aureus. This report is similar to the
bioactivity was found to be similar. findings of Ishikawa et al (2001), who
Bioactivity per gram was therefore showed that the mycelial culture filtrate of
increased when maltose was utilized as the Lentinula edodes inhibited the growth of
carbon source. It is possible that fructose B. subtilis. The observed inhibitory effect
as a simple sugar interferes with product of Pleurotus ostreatus on both gram
formation and maltose as a slow releasing negative and gram positive bacteria is in
carbon sources supported production of line with the report of Komemushi et al
bioactive compounds rather than growth of (1995, 1996) who studied the
the fungus. Also, with the exception of the antimicrobial substances in L. edodes.
medium containing glucose, none of the Imtiaj and Lee (2007) also worked on the
other cultures totally inhibited spore antibacterial and antifungal activities of
germination of test organism. Sucrose did Korean wild mushrooms and found that
not support growth of the fungus and several filtrates of wild mushrooms
consequently had no effect on production inhibited the growth of many pathogenic
of bioactive compounds. The same result bacteria such as P. aeruginosa and S.
has also been reported that sucrose is a aureus. The metabolite showed
very poor carbon sources for growth of

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antagonistic effect against all tested

pathogens and slightly affected P. Finally, this study has revealed the
aeruginosa, with in parallel with the antimicrobial activity of the edible
results of Adebayo et al ( 2012) .The mushrooms under study and can be
ability of the metabolite to inhibit all suggested that the bioactive contents of the
tested organisms with slightly inhibition to mushrooms are promising natural
P. aeruginosa, suggests that this product antimicrobial agents that can be harnessed
contained potential antibacterial agents as potential antibacterial and fungi
against infections from these pathogens. toxicants. Further extensive studies are
recommended for this mushroom and
The obtained results are in agreement with other mushrooms types to actually identify
the results reported by Ramseur et al the bioactive components responsible for
(2003) who investigated that crude extract their antimicrobial activities.
of African mushroom, P.tuberregium had
antibacterial activity against several food References
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P. tuber-regium extract had powerful antimicrobial assay of mushroom
medicinal importance by inhibiting the metabolite from Pleurotus
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