Evaluation of Antimicrobial, Antioxidant, Cytotoxic and DNA Protective Effects of Oyster Mushroom: Pleurotus pulmonarius (Fr.) Quel.

Abstract

Due to climate change and economic difficulties, food production and supply in the world is getting harder and harder. The importance of edible mushrooms is increasing day by day due to their consumption as food, nutritional value and health benefits. Many types of mushrooms are cultivated to overcome the difficulties of obtaining them naturally. Among the mushrooms cultivated in the world, the Pleurotus species ranks second after Agaricus spp. The aim of the current research was to determine the antimicrobial, antioxidant, cytotoxic and deoxyribonucleic acid protective effects of Pleurotus pulmonarius (Fr.) Quel. grown on some local cellulosic wastes. The antimicrobial activity and the minimum inhibitory concentration (MIC) tests carried out by the disk diffusion method and macro-broth dilution techniques. Antioxidant activity was ascertained by the radical scavenging capacity method of 2.2-diphenyl-1-picrylhydrazil. Total antioxidant status and total oxidant status of P. pulmonarius extracts were determined with rel assay kits. The DNA protective effect was evaluated using plasmid pBR322 DNA treated with ultraviolet (UV) and H2O2, and also the cytotoxic effects on the human lung cancer cell (A549) and human breast cancer cell (MDA-MB-231) lines were evaluated using the 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay method. It was observed to be highly active against all pathogen bacteria (except S. mutans) and dermatophytes (Epidermophyton sp. and Trichophyton sp.) (14.0–29.3 mm) compared to the control group (10.0–32.7 mm), but low against Candida tropicalis. It also showed significant antioxidant potential, the highest TAS (1.33 mmol/L) and TOS (6.74 µmol/L) values were obtained in the mixture of Medicago sativa + Populus nigra (MS-PS (1:1)), on Medicago sativa (MS) medium, and also DPPH radical scavenging effect was more effective at 25 mg (68.09%) on MS-PS (1:1) medium. The methanol extract of P. pulmonarius cultured on MS-PS (1:1) at a concentration of 400 μg/ml significantly reduced the percent viability in MDA-MB-231 cell lines (1.41%). However, it was determined that P. pulmonarius did not have a DNA protective effect. As a result, it was observed that P. pulmonarius had strong antimicrobial, antioxidant and cytotoxic activities, but did not have a DNA protective effect.

Full text

Arabian Journal for Science and Engineering (2023) 48:7273–7283
https://doi.org/10.1007/s13369-022-07418-9
RESEARCH ARTICLE-BIOLOGICAL SCIENCES
Evaluation of Antimicrobial, Antioxidant, Cytotoxic and DNA Protective
Effects of Oyster Mushroom: Pleurotus pulmonarius (Fr.) Quel.
Mehmet Akyüz1·¸Sule ˙
Inci2·Sevda Kırba ˘g2
Received: 9 March 2022 / Accepted: 23 October 2022 / Published online: 4 November 2022
© King Fahd University of Petroleum & Minerals 2022
Abstract
Due to climate change and economic difficulties, food production and supply in the world is getting harder and harder.
The importance of edible mushrooms is increasing day by day due to their consumption as food, nutritional value and
health benefits. Many types of mushrooms are cultivated to overcome the difficulties of obtaining them naturally. Among
the mushrooms cultivated in the world, the Pleurotus species ranks second after Agaricus spp. The aim of the current
research was to determine the antimicrobial, antioxidant, cytotoxic and deoxyribonucleic acid protective effects of Pleurotus
pulmonarius (Fr.) Quel. grown on some local cellulosic wastes. The antimicrobial activity and the minimum inhibitory
concentration (MIC) tests carried out by the disk diffusion method and macro-broth dilution techniques. Antioxidant activity
was ascertained by the radical scavenging capacity method of 2.2-diphenyl-1-picrylhydrazil. Total antioxidant status and
total oxidant status of P. pulmonarius extracts were determined with rel assay kits. The DNA protective effect was evaluated
using plasmid pBR322 DNA treated with ultraviolet (UV) and H2O2, and also the cytotoxic effects on the human lung
cancer cell (A549) and human breast cancer cell (MDA-MB-231) lines were evaluated using the 3-(4,5-Dimethylthiazol-2-
yl)-2,5-diphenyltetrazolium bromide (MTT) assay method. It was observed to be highly active against all pathogen bacteria
(except S. mutans) and dermatophytes (Epidermophyton sp. and Trichophyton sp.) (14.0–29.3 mm) compared to the control
group (10.0–32.7 mm), but low against Candida tropicalis. It also showed significant antioxidant potential, the highest TAS
(1.33 mmol/L) and TOS (6.74 μmol/L) values were obtained in the mixture of Medicago sativa +Populus nigra (MS-PS
(1:1)), on Medicago sativa (MS) medium, and also DPPH radical scavenging effect was more effective at 25 mg (68.09%)
on MS-PS (1:1) medium. The methanol extract of P. pulmonarius cultured on MS-PS (1:1) at a concentration of 400 μg/ml
significantly reduced the percent viability in MDA-MB-231 cell lines (1.41%). However, it was determined that P. pulmonarius
did not have a DNA protective effect. As a result, it was observed that P. pulmonarius had strong antimicrobial, antioxidant
and cytotoxic activities, but did not have a DNA protective effect.
Keyword Pleurotus pulmonarius ·Medicinal effects ·Edible mushroom ·Cultivation
1 Introduction
For thousands of years, mushrooms have been recognized
as suitable for human consumption and traditional treat-
ment [1]. They have been consumed since ancient times for
their nutritional benefits as well as their medicinal values
[2]. Historically, awareness of the medicinal properties of
BMehmet Akyüz
makyuz@beu.edu.tr
1Department of Biology, Faculty of Arts and Science, Bitlis
Eren University, 13000 Bitlis, Turkey
2Department of Biology, Faculty of Science, Fırat University,
23119 Elazı˘g, Turkey
mushrooms comes from the folklore of different continents
such as Asia, Central Europe, South America and Africa [3].
However, awareness of mushrooms as an important source
of biologically active substances with medicinal value has
only recently emerged and they are used as herbal medicines
[4].
The genus Pleurotus species, which are commonly
regarded as oyster mushrooms, are edible mushroom from
the well-known genus of Basidiomycota. They are the second
most widely cultivated genus in the world after Agari-
cus bisporus (J.E. Lange) Imbach due to their adaptability.
This genus includes many white rot fungi with significant
culinary interest, medicinal properties, potential biotech-
nological and environmental applications [5–7]. Different
123

7274 Arabian Journal for Science and Engineering (2023) 48:7273–7283
species of Pleurotus are widely cultivated in Southeast Asia,
India, Europe, American continent and Africa, with approx-
imately 200 species found worldwide in both temperate and
tropical regions [8,9]. In recent years, interest in Pleuro-
tus spp., which is among the most popular mushroom types
on the world’s surface, has been increasing. The main rea-
son for this is that they have different species, easy culture
conditions, rich nutritional content and many bioactive com-
ponents they contain have different medicinal effects. They
contain components such as polysaccharides (α-glucans and
β-glucans), proteins (such as eryngin and pleurostrin), glyco-
proteins, lectins, high molecular weight primary metabolites,
low molecular weight secondary metabolites (alkaloids, beta-
lains, fatty acids and esters, flavonoids (such as chrysin,
myricetin, naringenin, quercetin and rutin), polyphenols
(caffeic, cinnamic, chlorogenic, p-coumaric, ferulic, gallic
and protocatechuic acids) and triglyceridesols which have
important medicinal effects [5,10–21]. These bioactive
components of Pleurotus spp. show antiaging, antialler-
gic, anticancer, antigenotoxic, anti-inflammatory, antihyper-
glycemic, antihypertensive, antimicrobial, antimutagenic,
antioxidant, antitumor, antiviral, cytotoxic, hematologi-
cal, hepatoprotective, hypocholesterolemic, hypolipidemic,
DNA protective effects and immunomodulatory effects [1,
13,15,22–28].
Globally, we encounter three different types of disease
on the earth’s surface, often caused by microbial pathogens,
free radicals and cancer, among other diseases. Infectious
diseases remain one of the major threats to human health
throughout the world. In the developed world, infectious
diseases including bacterial food poisoning, gonorrhea, coli-
tis, chlamydia, acute gastroenteritis, urinary tract infections,
bloodstream infection, joint infection, bacterial cellulitis and
vaginosis, meningitis, syphilis, strep throat, respiratory tract
infection, etc., represent the largest proportion of diseases
today [29]. In addition to all these, oxidative stress caused
by free radicals can be associated with serious nerve prob-
lems, aging, cirrhosis, diabetes, atherosclerosis and cancer
[30–32]. Free radicals form in cell membranes and nuclei
and damage lipids, proteins, nucleic acids and carbohydrates,
and they also have the capacity to attack vital macro-
molecules, damage cells and tissues, disrupt homeostasis and
metabolic processes [16]. They are produced not only within
the organism but also by various external sources includ-
ing ultraviolet light, ionizing radiation, environmental toxins,
chemotherapeutics and inflammatory cytokines [17]. Due to
the increasing diseases in the world, the need for medicine is
increasing day by day. In order to minimize the side effects
of drugs used in the treatment of diseases, the demand for
natural products is increasing. The combination of these nat-
ural products with synthetic drugs, their use as a supplement
or their use alone are preferred. For this reason, it is one of
the natural agents that can be recommended in the treatment
of diseases in fungi. In this sense, considering its nutritional
and medicinal effects, the medicinal effects of P. pulmonarius
need to be further clarified. Although this species has been
studied by researchers, it is a species that is little known
by consumers. Therefore, before the use of this species, its
medicinal effects should be well known.
Pleurotus pulmonarius (Fr.) Quel. belongs to the class
Agaricomycetes and is also known as “Indian Oyster, Italian
Oyster, Phoenix Mushroom or Lung Oyster.” Its fruiting bod-
ies are characterized by a mild taste and a slight anise aroma.
P. pulmonarius is valued as a source of nutrients and sub-
stances with a healing effect [33,34]. It has been determined
that P. pulmonarius contains monounsaturated fatty acids,
oleic acid [35] and β-glucan, which has anti-inflammatory
and analgesic properties [36], sugar (xylose, glucose, ara-
binose), amino acid (aspartic acid, glycine, arginine, lysine,
threonine, glutamic acid, tyrosine, alanine and valine), and
polar lipid (diphosphatidylglycerol, phosphatidylglycero,
phosphatidyl-N-methylethanolamine, phosphatidylcholine,
phosphatidylserine, phosphatidylethanolamine, unidentified
phospholipid, unidentified aminolipid, unidentified glycol-
ipid, unidentified lipid) [37]. It is also a good source of
protein, energy and carbohydrates [38]. Thanks to the bio-
chemical ingredients, P. pulmonarius is confirmed to possess
several therapeutic properties such as anti-diabetic, antiox-
idative, anti-inflammatory and hypolipidaemic [39]. In nat-
ural conditions, P. pulmonarius is most commonly found in
North America [40]. Researchers stated that this species can
be grown on wastes such as cocoa shells, cotton waste, corn
straw, palm oil waste, tobacco straw, tea leaves, rice straw,
sugarcane pomace, waste paper, sawdust and other substrates
[38,40–43].
Today, it has become increasingly important to identify
new fungal-derived metabolites that can be used as multi-
spectral therapeutics. The various features observed continue
to make edible mushrooms promising candidates for biolog-
ical research activities. Therefore, prospective studies that
will reveal its potential medicinal properties could be an asset
for the global health industry [21]. Considering all these, the
aim of the current research is to determine the possible antiox-
idant, antimicrobial, cytotoxic and DNA protective effect of
P. pulmonarius.
2 Material and Methods
2.1 Obtained of Mushroom Samples
The main culture of Pleurotus pulmonarius (Fr.) Quel. was
obtained from the Department of Biology, Science Faculty,
Fırat University, Elazı˘g-Turkey and maintained on potato
dextrose agar medium at 4 °C. For inocula multiplication,
propagation of spawn, cultivation process such as substrate
123

Arabian Journal for Science and Engineering (2023) 48:7273–7283 7275
preparation, inoculation of substrates, maintenance of beds
and for harvest, the methods proposed by Zadrazil [44]were
followed. For the formation of basidiocarp, Medicago sativa
L. (MS), Prangos pabularia Lindl. (PP) and Populus nigra L.
(Poplar sawdust) (PS) substrates were used as culture media.
Three types of compost were prepared, consisting of a mix-
ture of MS-PP (1:1), MS-PS (1:1) and MS with 70–75%
humidity. In addition, MS was used as the control treatment.
The samples used in the current research were taken from
the cultural studies conducted by Akyüz et al. [38]. Fresh
mushroom samples obtained from the culture medium were
dried in the shade for 10 days, pulverized and immediately
analyzed for use in other steps.
2.2 Extraction of Mushroom Samples
Mushroom was dried in a dry air sterilizer at 40 °C for
10–12 h. They were pulverized in a blender. 50 g of pow-
dered samples was left in sterile flasks and 100 ml of
96% methyl alcohol (MetOH) was added separately. These
extracts, which were prepared separately for each of the
mushroom samples, were processed in a shaking oven for
48 h, the solvents of the extracts were removed in the rotava-
por and dried in the oven at 40 °C to obtain dry extracts. All
extracts prepared in this way were stored in the refrigerator at
4 °C until the analysis and the conclusion of the experiment
[45].
2.3 Antimicrobial Activity of Mushroom Samples
2.3.1 Disk Diffusion Method
A total of 6 bacteria (Pseudomonas aeruginosa DMS 50,071
SCOTTA, Escherichia coli ATCC 25,922, Staphylococcus
aureus COWAN 1, Streptococcus mutans,Proteus vulgaris
FMC1, Klebsiella pneumoniae ATCC 700,603), 1 yeast
(Candida tropicalis ATCC 13,803) and 2 dermatophyte (Epi-
dermophyton sp. and Trichophyton sp.) species were used in
this study. Microorganisms were provided by the Microbiol-
ogy Research Laboratory, Department of Biology, Faculty of
Science, Firat University, Elazig-Turkey. The antimicrobial
tests were carried out by the disk diffusion method [46]using
100 μL of suspension containing 106per/mL of bacteria,
104per/mL yeast and dermatophyte inoculated into Mueller
Hinton Agar (Difco), Malt Extract Agar (Difco) and Glukoz
Sabouroud Agar (Difco), respectively. The disks (6 mm)
were then impregnated with 40 μL (200 μg) of mushroom
extract and then placed on the inoculated agar. Petri dishes
were prepared at 4 °C for 2 h. Then, the inoculated plates
were incubated at 37 ±0.1 °C for 24 h for bacterial strains
and also 25 ±0.1 °C for 72 h for yeast and dermatophyte.
Streptomycin sulfate (10 μg/disk) for bacteria and nystatin
(30 μg/disk) for yeast and dermatophyte were used as the
positive control and methanol as the negative control. At the
end of the incubation period, the inhibition zones were mea-
sured.
2.3.2 Minimum Inhibitory Concentration
The minimum inhibitory concentration (MIC) of MetOH
extracts of mushrooms was detected using macro-broth
dilution techniques [47]. A twofold serial dilution of the
reconstituted extract was prepared in Mueller Hinton Broth.
Each dilution was seeded with 100 μl of the standardized
suspension of the test organisms and incubated for 24 h at
37 °C 24 h for bacterial strains and also 25 ±0.1 °C for
72 h for yeast and dermatophyte. All extracts were tested
at 500–15.625 μg/mL concentrations. MIC was determined
as the highest dilution (that is, lowest concentration) of the
extract that showed no visible growth [47].
2.4 Antioxidant Activity of Mushroom Samples
2.4.1 Total Antioxidant Activity and Total Oxidant Activity
Total antioxidant status (TAS) and total oxidant status (TOS)
of mushroom extracts were determined with Rel Assay
kits (Rel Assay Kit Diagnostics, Turkey). TAS value was
expressed as mmol Trolox equiv./L and Trolox was used as
the calibrator. The TOS value was expressed as μmol H2O2
equiv./L and hydrogen peroxide was used as the calibrator
[48,49].
2.4.2 DPPH Assay
The antioxidant activity of the different concentrations of the
MetOH extracts of mushroom were determined according to
the 2.2-diphenyl-1-picrilhydrazyl (DPPH) radical scaveng-
ing capacity method [50,51]. The solution was prepared
in methanol at a concentration of 25 mg/ml of the extract
obtained. The prepared solution was diluted four times and
the calibration curve of DPPH was obtained. By taking 40 μl
of the prepared solution, 160 μl of DPPH solution was added.
After thorough mixing, the mouth was closed and kept in
the dark for 30 min. The same procedures were repeated
for all concentrations and methanol was used as a control.
At the end of this period, the absorbances of each mix-
ture were read at 570 nm in the elisa reader. % inhibition
values were calculated by the following equation: I%=
[(AControl—ASample)/AControl]×100.
2.5 Cytotoxic Activity
The human lung cancer cell (A549) and human breast cancer
cell (MDA-MB-231) lines used in this study were obtained
123

7276 Arabian Journal for Science and Engineering (2023) 48:7273–7283
Fig. 1 Fructification of P.
pulmonarius (a—grown on
culture medium, b—basidiocarp)
from Inonu University, Turkey. The 3-(4,5-Dimethylthiazol-
2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay method
was used to detect the cytotoxic effect of methanol extracts
of mushrooms [52]. The incubated MDA-MB-231 and
A549 cell lines were removed from culture plates by using
trypsin–EDTA solution and inoculated into 96-well (15 ×
103cells per well) culture plates. These cell culture plates
were incubated at 37 °C for 24 h. After incubation, 100 μl
from different concentrations (200 μg/ml and 400 μg/ml) of
methanol extracts of mushroom were added and incubated
for24hat37°Cin5%CO
2incubator. At the end of the incu-
bation, 20 μl of MTT solution was added and at the end of
4 h incubation, absorbance measurements at 570 nm wave-
lengths were made. Doxorubicin was used as positive control
and DEMEM as a negative control.
2.6 DNA Protective Activity
DNA damage protection activities of the extracts were eval-
uated on pBR322 plasmid DNA (vivantis). Plasmid DNA
was oxidized with H2O2+UV treatment in the presence
of extracts and checked on 1.25% agarose gels according
to literature [53,54] after some modifications. In brief,
the experiments were performed in a volume of 10 μlin
a microfuge tube containing 3 μl pBR322 plasmid DNA
(172 ng/μl), 1 μl of 30% H2O2and 5 μl of extract in
the concentration of 25 mg/ml. The reactions were initiated
by UV irradiation and continued for 5 min on the surface
of a UV transilluminator (DNR-IS) with an intensity of
8000 μW/cm2at 302 nm at room temperature. After irradia-
tion, the reaction mixture (10 μl) along with gel loading dye
(6 ×) was loaded on a 1.25% agarose gel for electrophoresis.
Untreated pBR322 plasmid DNA was used as a control in
each run of gel electrophoresis along with partially treated
plasmid, i.e., only UV or only H2O2treatment. Gels were
stained with EtBr and photographed with the Gel documen-
tation system (DNR-IS, MiniBIS Pro) [53,54].
2.7 Statistical Analysis
Data were represented as mean ±standard deviation (SD)
based on at least three replicates, and the significant dif-
ference (p< 0.05) was determined by one-way analysis of
variance with Duncan’s test using SPSS v25.0.
3 Result and Discussion
In this study, the methanol extract of P. pulmonarius grown
on a consisting of a mixture of MS-PP (1:1), MS-PS (1:1)
and MS (Fig. 1) were investigated for biological activities at
various concentrations (see Tables 1,2,3,Fig.2).
Fruiting bodies and mycelium of mushroom have com-
ponents with broad antimicrobial effects, so they are a good
source of active biological compounds. Antimicrobial activ-
ity of methanol extracts of P. pulmonarius grownonsome
cellulosic wastes were tested in detail against several bacte-
ria (P. aeruginosa,E. coli,S. aureus,S. mutans,P. vulgaris
and K. pneumoniae), yeast (C. tropicalis) and dermatophytes
(Epidermophyton sp. and Trichophyton sp.) using disc diffu-
sion method (see Table 1), and the zones of inhibition were
observed and the results are showed in Table 1. Methanol
extract of P. pulmonarius was observed to be changeable
statistically against bacteria, yeast and dermatophyte when
compared with the control (see Table 1,p< 0.05). The results
are presented in Table 1and indicate that mushroom extracts
revealed to be highly active against all bacteria (except S.
mutans) and dermatophytes (Epidermophyton sp. and Tri -
chophyton sp.) (14.0–29.3 mm) compared to the control
group (10.0–32.7 mm), but lower against yeast (C. tropi-
calis). Also, the antimicrobial activities of P. pulmonarius
123

Arabian Journal for Science and Engineering (2023) 48:7273–7283 7277
Table 1 Antimicrobial effects (mm) and MIC test (μg/mL) of P. pulmonarius cultured on various local cellulosic wastes
Compost
Medium
(1:1)
Bacteria Yeast Dermatophytes
Gram (+) Gram (−)
S. mutans S. aureus P. vulgaris P. aeroginosa K. pneumoniae E. coli C. tropicalis Epidermophyton sp. Trichophyton sp.
MIC MIC MIC MIC MIC MIC MIC MIC MIC
MS 23.3±0.6a– 25.7±0.6c31.25 20.0±0.0c– 20.3±0.6b31.25 20.3±0.6b31.25 24.0±1.0b62.50 21.7±0.6c– 25.7±0.6c31.25 20.0±0.0c31.25
MS-PS 22.7±1.5a– 19.7±0.6b62.50 18.3±0.6b31.25 20.0±0.0b62.50 17.7±0.6a62.50 29.3±1.2c62.50 17.7±0.6b31.25 26.7±1.2c62.50 19.3±1.2c62.50
MS-PP 21.7±1.5a– 19.7±0.6b31.25 18.0±0.0b31.25 20.6±0.6b62.50 18.3±0.6a62.50 23.3±0.6b62.50 14.0±0.0a–20.0±0.0b62.50 14.3±0.6b62.50
Control 32.7±0.6b14.7±0.6a11.3±0.6a10.3±0.6a18.0±0.6a10.0±0.0a29.3±1.2d14.7±0.6a11.7±0.6a
Fvalue 58.562 182.250 263.167 300.889 12.000 304.667 259.111 185.500 96.444
pvalue 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000
Values are means of three replicates ±SD, Means in the same column with the different superscript are significantly different (p< 0.05), a.b.c: Comparison in different culture medium, MIC—Minimal
inhibitor concentration, Control groups for bacteria (Streptomycin sulfate (10 μg/disk)), yeast and dermatophyta (Nystatin (30 μg/disk)), MS—Medicago sativa L., PP—Prangos pabularia Lindl.,
P—Poplar sawdust
123

7278 Arabian Journal for Science and Engineering (2023) 48:7273–7283
Table 2 TAS, TOS values and
DPPH of P. pulmonarius cultured
on some local lignocellulosic
wastes
Materials
(1:1)
TAS
(mmol/L)
TOS
(μmol/L)
DPPH (%)
25 mg/mL 12.5 mg/mL 6.25 mg/mL 3.125 mg/mL
MS 0.83 6.74 63.21 17.89 6.10 1.10
MS-PS 1.33 3.67 68.09 32.11 10.46 3.49
MS-PP 0.58 2.58 42.48 13.82 10.47 4.37
PS—Poplar sawdust, MS—Medicago sativa L., PP—Prangos pabularia Lindl
TAS—the total antioxidant status, TOS—total oxidant status, DPPH—2.2-diphenyl-1-picrilhydrazyl
against S. mutans,S. aureus,P. vulgaris,P. aeroginosa were
not statistically different in all compost medium, whereas
against P. vulgaris,K. pneumonia,E. coli,Epidermophyton
sp. and Trichopyton sp. were found to differ (Table 1). In
addition, minimal inhibitor concentration (MIC) may vary
for the studied species (31.25–62.5 μg/mL) shown Table 1.
Fungi can form excellent sources of substitutes for
bioactive compounds with antimicrobial properties, pre-
dominantly secondary metabolites such as phenolic com-
pounds, flavonoids, vitamins, carotenoids, alkaloids, glyco-
sides, steroids, terpenoids, enzymes, organic acids, as well
as several primary sources of metabolites, including polysac-
charides, peptides and oxalic acid [1,5,10–29,55–58]. Alves
et al. [57] suggested that the edible mushrooms have antimi-
crobial activity against certain Gr (−) and Gr (+) bacterial
pathogens. Parihar et al. [58] stated that mushroom com-
pounds showed higher antibacterial effect against Gr (+)
bacteria than Gr (−) bacteria. Chang and Wasser [55] and
Patel and Goyal [56] indicated that polysaccharides display
important medical features, and that β-glucan is the most
popular and flexible metabolite with a wide variety of natu-
ral interactions. In the previous reports [1,5,10–29,55–59],
there were several data for the antimicrobial activities of edi-
ble wild, cultured and commercially mushroom. The results
reported by different authors above are difficult to com-
pare, due to the variety of methodologies used to evaluate
the antimicrobial activity of mushroom extracts or isolated
compounds. Contrary to our results, multiple investigators
found antimicrobial activity values for different mushroom
species. In the current research, the methyl alcohol extracts
of P. pulmonarius exhibited different levels of antimicrobial
activity than previously reported for other extracts. Similar
studies mentioned above also showed a significant relation-
ship between fungal bioactive compounds and antimicrobial
effect. The intensity of the antimicrobial effect will vary
depending on the type of fungus, suitable solvents and their
concentration, the methods used and organism tested. The
differences in the antimicrobial effect of the tested fungus are
probably the result of the presence of different components
with antimicrobial as indicated in the mentioned studies.
Table 3 Cytotoxic effect of P. pulmonarius extracts on A-549 and
MDA-MB-231 cell lines (%)
Concentration Materials (1:1) Cell line
A-549 MDA-MB-231
200 μg/ml MS 93.69 38.11
MS-PS 89.50 32.93
MS-PP 87.34 35.57
400 μg/ml MS 13.00 2.35
MS-PS 9.38 1.41
MS-PP 14.07 5.29
Positive control (Doxorubicin) 49.20 10.69
Negative control (DEMEM) 100 100
PS—Poplar sawdust, MS—Medicago sativa L., PP—Prangos pabu-
laria Lindl
A-549—Human lung cancer cell line, MDA-MB-231—human breast
cancer cell line
The antioxidant effects have been attributed to variety
reactions and mechanisms, including binding of transi-
tion metal ion catalysts, β-carotene-linoleic acid, reductive
capacity, xanthine oxidase, radical scavenging, ferrous ions
chelating abilities and prevention of chain initiation, etc. [16].
Hydroxyl radicals are the main reactive oxygen species caus-
ing biological damage and lipid peroxidation [60]. Other
methods including superoxide anion radical and hydroxyl
radical scavenging activity are used to know and under-
stand the antioxidant action mechanism [61]. However, the
DPPH test is used as a simple, quick, reproducible and main
test in antioxidant effects researches, so it is widely used
to predict the antioxidant activities of compounds [62,63].
In the current study, the in vitro antioxidative capacities of
P. pulmonarius were evaluated using different biochemical
methods such as total antioxidant status (TAS), total oxidant
status (TOS), as well as DPPH scavenging assay, as seen in
Table 2. The results of TAS, TOS and DPPH scavenging assay
are described in Table 2. Table 2showed that the TAS and
TOS values of P. pulmonarius vary depending on the culture
medium. However, it was determined that the total antiox-
idant levels of P. pulmonarius grown on MS, MS-PS (1:1)
123

Arabian Journal for Science and Engineering (2023) 48:7273–7283 7279
and MS-PP (1:1) were low, but the total oxidant levels were
very good. DPPH radical scavenging effects were more effec-
tive on MS-PS (1:1) at 25 mg/ml (68.09%), respectively (see
Table 2). DPPH of P. pulmonarius varied from 1.10 to 68.09%
and increased with increase in concentration (show Table 2).
Antioxidant has the ability to either destroy or reduce free
radicals. They scavenge free radicals leading to inhibition of
oxidative mechanisms that cause various degenerative dis-
eases. Dulay et al. [64] noted that DPPH is a simple method
developed to determine the antioxidant activity of any poten-
tial substance or compound. Zhang et al. [65] worked that the
natural selenium-enriched polysaccharide fraction produced
by Pleurotus ostreatus (Jacq.) P. Kumm. showed strong scav-
enging capacity against DPPH and hydroxyl radical. Sudha
et al. [66] noted that Pleurotus djamor (Rumph. ex Fr.)
Boedijn extracts scavenge the stable DPPH radical to vary-
ing degrees (7.40–85.19%). Menaga et al. [1] reported that
the methyl alcohol extract of P. ostreatus seen the highest
DPPH scavenging activity (76%). Song et al. [67]showed
that polysaccharides can cause a cycle of antioxidant activ-
ity in cells, inhibit ROS production, regulate the cell cycle,
prevent DNA damage and protecting cell membrane. Liu
et al. [63] indicated that the intracellular polysaccharide from
Pleurotus eryngii (DC. ex Fr.) Quel., Pleurotus cornucopiae
(Paulet) Quél. and Pleurotus nebrodensis (Inzenga) Quél. can
effectively protect cell from damage and lipid peroxidation.
The in vitro DPPH radical scavenging assay of polysaccaride
of Pleurotus spp. were 17.9%, 16.8% and 20.5%. Kim et al.
[68] suggested that naringin, myricetin and phenolic acids
in Pleurotus spp. have antioxidant, anti-inflammatory and
antitumor properties. Finimundy et al. [69] stated that phe-
nolic compounds known to have strong antioxidant activity
were reported in mushrooms. Sudha et al. [66] and G˛asecka
et al. [70] observed a positive correlation between phenolic
content in macrofungi and the antioxidant activity by DPPH
radical scavenging. However, Viera et al. [71] found no such
relationship between phenolic compounds and antioxidant
activity. Therefore, it was stated that the differences in the
bioavailability of the enriched elements may be related to
the antioxidant activity differences observed with the DPPH
method. These data of Table 2showed that the P. pulmonarius
could effectively protect the cell from damage and lipid per-
oxidation. Compared with those studies, mushroom strain
showed remarkable DPPH free radical scavenging activi-
ties under the selected concentration, and many phenols,
flavonoids and related compounds found in fungal structures
were reported to have potent antioxidative properties in the
above-mentioned studies. The intensity of antioxidant activ-
ity may vary depending on the fungal species tested and the
solvent used for extraction using different methods. Differ-
ences in antioxidant effects of various solvents may be the
result of different ability to extract bioactive substances.
Fig. 2 Electrophoretic pattern of pBR322 plasmid DNA after treatment
with dH2O, UV and H2O2in the presence of water extracts [K1: Plas-
mid DNA (3 μl) +dH2O(6μl), K2: Plasmid DNA (3 μl) +dH2O
(6 μl) +H2O2(1 μl) +UV, 1: Plasmid DNA (3 μl) +25 mg/ml of
P. pulmonarius (MS) methyl alcohol extracts +UV +H2O2(1 μl),
2: Plasmid DNA (3 μl) +25 mg/ml of P. pulmonarius (MS-PS (1:1))
methyl alcohol extracts +UV +H2O2(1 μl), 3: Plasmid DNA (3 μl)
+25 mg/ml of P. pulmonarius (MS-PP (1:1)) methyl alcohol extracts
+UV +H2O2(1 μl), OX: Plasmid DNA (3 μl) +Oxibenzon (5 μl)
+UV +H2O2(1 μl), respectively]
A variable cytotoxic effects on human lung cancer cell
lines (A549) and human breast cancer cell lines (MDA-MB-
231) were observed at concentrations of 200–400 μg/ml of
the methanol extract of P. pulmonarius cultured on vari-
ous local cellulosic residues (see Table 3). Methyl alcohol
extracts of P. pulmonarius were shown to be variable against
cell lines A-549 and MDA-MB-231 when compared to neg-
ative (DEMEM) and positive (doxorubicin) control groups
(show Table 3). The methanol extracts of P. pulmonar-
ius showed low cytotoxic activity on A-549 cell line at
200 μg/mL, but showed high cytotoxic activity on MDA-
MB-231 cell line at 400 μg/mL (see Table 3). Consequently,
P. pulmonarius cultured on MS-PS (1:1) at a concentra-
tion of 400 μg/ml significantly reduced the percent viability
in MDA-MB-231 cell lines (1.41%) as seen in Table 3.
Reactive oxygen species play important roles in patholog-
ical processes such as coronary heart disease, Alzheimer’s
disease, inflammation, severe neurodegenerative disorders,
cataract formation, cancer and aging [30–32,72]. How-
ever, over production of reactive oxygen species (ROS) can
cause oxidative damage to the function of macromolecules,
DNA, genomic instability and proteins. Excessive accumu-
lation of ROS promotes the acquisition of mutations and
may eventually cause cellular functional changes in cancer
cells [16]. Mariga et al. [3] stated that a novel bioactive pro-
tein extracted from P. eryngii significantly inhibited human
cell lines (HGC-27), human gastric cancer cell line (BGC-
823), human lung cancer cell lines (A549) and human
hepatoma cells (HepG2) tumor cells. Ma et al. [73] noted
that polysaccharides extracted from P. eryngii residue exhib-
ited high cytotoxicities toward HepG-2 cells. Khan et al. [74]
revealed that P. ostreatus showed the highest cell prolifera-
tion inhibition against human colon cancer cells (Colo-205)
123

7280 Arabian Journal for Science and Engineering (2023) 48:7273–7283
and human breast cancer (MCF-7) cell. Sun et al. [75]
reported that selenium-enriched Drechmeria gunnii (Berk.)
Spatafora, Kepler & C.A. Quandt could influence the cell
viability of HepG2 cells, human ovarian cancer (SKOV3)
cells and human lung cancer cell lines (H1299 cells) in a
dose-dependent and time-dependent manner. Maity et al. [76]
observed a novel substance β-glucan from P. djamor showed
that high cytotoxic activity against ovarian teratocarcinoma
(PA-1) cell lines. EL-Deep et al. [20] indicated that P. ostrea-
tus polysaccharides induce NK-cell cytotoxic effects against
HepG2, MCF7 and A549 cells, with the greatest effect
against MCF7 cells (81.2%). Zhang et al. [65] showed that the
selenium-enriched polysaccharide fraction produced by P.
ostreatus could reduce the viability of A549, SKOV3, HepG2
and MCF-7 cells, induce apoptosis and inhibit metastasis of
A549 cells, but no significant effect on normal cells. Menaga
et al. [1] found that 3-methoxy-4-hydroxy cinnamic acid
from the methanolic extract of P. ostreatus has a remarkable
effect against A549 cells. Elhusseiny et al. [77], Agaricus bis-
porus (J.E. Lange) Imbach, P. ostreatus and P. pulmonarius
extracts showed low cytotoxicity against PBMCs and moder-
ate cytotoxicity against hepatocellular carcinoma (HepG2),
colorectal carcinoma (Colo-205), human lymphoma (U937),
human prostate cancer (PC3 and DU-145), breast adeno-
carcinoma (MCF-7 and MDA-MB-231), human leukemia
(CCRF-CEM), cecum carcinoma (LS-513), acute monocytic
leukemia (THP1), acute promyelocytic leukemia (NB4) and
cervical cancer (HeLa) cell lines. Considering all these, it
has been proven that many bioactive components found in
fungal structures have cytotoxic effects [18–21]. The data we
obtained in Table 3may differ from the data in the mentioned
studies [1,3,65,73–77]. The main reason for the difference
in data obtained may be due to different analytical methods,
using chemicals, mushroom species, cancer cell line tested
and heterogeneity of samples.
As seen in Fig. 2, DNA preserved its stable structure in
the lanes with K1 and Oxybenzone. The superhelical struc-
ture of DNA can be seen in these lanes. In the K2 lane, DNA
could not maintain its stable structure under the influence
of UV and H2O2and a smear image was formed. Although
the DNA stable structure is preserved very little in the 1st
and 2nd lanes, it has been determined that it does not have a
DNA protective effect since the DNA cannot protect the sta-
ble structure in the 3rd lane. According to our results, methyl
alcohol extract of P. pulmonarius culturedonMS,MS-PS
(1:1) and MS-PP (1:1) substrates did not demonstrated a
protective effect to plasmid pBR322 DNA form the dam-
age induced by UV radiation and H2O2at a concentration
of 25 mg/ml (Fig. 2, Lane 1–3). It has been reported that
polysaccharide extracted from Ganoderma lucidum (Cur-
tis) P. Karst. fruiting bodies protects against DNA strand
breaks caused by hydroxyl radical [78]. Sa-Ard et al. [79]
reported that mycelium protein extract from G. lucidum had
higher potential DNA protective activities than fruiting bod-
ies protein extract. Akyüz et al. [80] investigated that Picoa
juniperi Vittad., only at a 40 mg/mL concentration demon-
strated a DNA protective effect, whereas Terfezia boudieri
Chatin, extract did not show any DNA protective effect at
all concentrations tested, and also higher concentrations of
Terfezia olbiensis Tul. & C. Tul. and Picoa lefebvrei (Pat.)
Maire extracts showed a DNA protective effect. However,
it was determined that P. pulmonarius did not have a DNA
protective effect (see Fig. 1). Edible wild, cultured and com-
mercial mushroom species have different components and
concentrations, which explains their different DNA protec-
tive activities as suggested in the studies mentioned above
[78–80]. In this respect, it may vary when compared to sim-
ilar studies.
4 Conclusion
Considering the results of the study, it was observed that P.
pulmonarius showed different levels of antimicrobial, antiox-
idant and cytotoxic effects, but did not have DNA protective
activity. Methy alcohol extract of P. pulmonarius grown on
MS medium revealed to be generally highly active against all
bacteria (except S. mutans) and dermatophytes (Epidermo-
phyton sp. and Trichophyton sp.) (14.0–29.3 mm) compared
to the control group (10.0–32.7 mm), but lower against
yeast (C. tropicalis). It was observed that the best DPPH
(25 mg/ml and 12.5 mg/ml), cytotoxic effect and TAS values
of P. pulmonarius grown on various local cellulosic wastes
were obtained from MS-PS (1:1) medium, but antimicro-
bial activities and TOS values were more effective in MS
medium, respectively. In addition, considering the richness
of the medicinal contents of P. pulmonarius, the use of MS
and MS-PS (1:1) wastes in the culture medium will be ben-
eficial.
Funding The authors declared that this study has received no financial
support.
Declarations
Conflict of interest The authors declare no competing interests.
Ethics Approval No ethical approval required.
References
1. Menaga, D.; Rahman, P.K.; Rajakumar, S.; Ayyasamy, P.M.:
Antioxidant and cytotoxic activities of a novel isomeric molecule
(PF5) obtained from methanolic extract of Pleurotus florida mush-
room. J. Bioresour. Bioprod. 6, 338–349 (2021)
123

Arabian Journal for Science and Engineering (2023) 48:7273–7283 7281
2. Smolskait˙e, L.; Venskutonis, P.R.; Talou, T.: Comprehensive eval-
uation of antioxidant and antimicrobial properties of different
mushroom species. LWT 60, 462–471 (2015)
3. Mariga, A.M.; Pei, F.; Yang, W.J.; Zhao, L.Y.; Shao, Y.N.;
Mugambi, D.K.; Hu, Q.H.: Immunopotentiation of Pleurotus eryn-
gii (DC. ex Fr.) Quel. J. Ethnopharmacol. 153, 604–614 (2014)
4. Rexhepi, B.; Reka, A.: Ethnomycological knowledge of some wild
medicinal and food mushrooms from Osogovo Mountains (North
Macedonia). J. Nat. Sci. Math. 5(9–10), 10–19 (2020)
5. Calabretti, A.; Mang, S.M.; Becce, A.; Castronuovo, D.; Cardone,
L.; Candido, V.; Camele, I.: Comparison of bioactive substances
content between commercial and wild-type isolates of Pleurotus
eryngii. Sustainability 13, 3777 (2021)
6. Karaman, M.; ˇ
Capelja, E.; Rašeta, M.; Raki´c, M.: Diversity, chem-
istry, and environmental contamination of wild growing medicinal
mushroom species as sources of biologically active substances
(Antioxidants, Anti-Diabetics, and AChE Inhibitors). In: Arya,
A.; Rusevska, K. (Eds.) Biology, Cultivation and Applications of
Mushrooms, pp. 203–257. Springer, Singapore (2022)
7. Melanouri, E.; Dedousi, M.; Diamantopoulou, P.: Cultivating
Pleurotus ostreatus and Pleurotus eryngii mushroom strains on
agro-industrial residues in solid-state fermentation. Part II: effect
on productivity and quality of carposomes. Carbon Resour. Con-
vers. 5(1), 52–60 (2022)
8. Bellettini, M.B.; Fiorda, F.A.; Maieves, H.A.; Teixeira, G.L.; Ávila,
S.; Hornung, P.S.; Ribani, R.H.: Factors affecting mushroom Pleu-
rotus spp. Saudi J. Biol. Sci. 26, 633–646 (2019)
9. Caldas, L.A.; Zied, D.C.; Sartorelli, P.: Dereplication of extracts
from nutraceutical mushrooms Pleurotus using molecular network
approach. Food Chem. 370, 131019 (2022)
10. Mattila, P.; Konko, K.; Eurola, M.; Pihlava, J.; Astola, J.; Vahter-
isto, L.; Hietaniemi, V.; Kumpulainen, J.; Valtonen, M.; Piironen,
V.: Contents of vitamins, mineral elements, and some phenolic
compounds in cultivated mushrooms. J. Agric. Food Chem. 49,
2343–2348 (2001)
11. Wang, H.; Ng, T.B.: Eryngin, a novel antifungal peptide from fruit-
ing bodies of the edible mushroom Pleurotus eryngii. Peptides
25(1), 1–5 (2004)
12. Valverde, M.E.; Hernández-Pérez, T.; Paredes-López, O.: Edible
mushrooms: improving human health and promoting quality life.
Int. J. Microbiol. 376387, 1–14 (2015)
13. Sharma, A.; Sharma, A.; Tripathi, A.: Biological activities of Pleu-
rotus spp. polysaccharides: a review. J. Food Biochem. 45(6),
e13748 (2021)
14. Torres-Martínez, B.M.; Vargas-Sánchez,R.D.; Torrescano-Urrutia,
G.R.; Esqueda, M.; Rodríguez-Carpena, J.G.; Fernández-López, J.;
Perez-Alvarez, J.A.; Sánchez-Escalante, A.: Pleurotus genus as a
potential ingredient for meat products. Foods 11(6), 779 (2022)
15. López-Hortas, L.; Flórez-Fernández, N.; Torres, M.D.;
Domínguez, H.: Update on potential of edible mushrooms:
high-value compounds, extraction strategies and bioactive
properties. Int. J. Food Sci. Tech. 57(3), 1378–1385 (2022)
16. Kehrer, J.P.; Klotz, L.O.: Free radicals and related reactive species
as mediators of tissue injury and disease: implications for Health.
Crit.Rev.Toxicol.45, 765–798 (2015)
17. Widel, M.; Krzywon, A.; Gajda, K.; Skonieczna, M.; Rzeszowska-
Wolny, J.: Induction of bystander effects by UVA, UVB, and UVC
radiation in human fibroblasts and the implication of reactive oxy-
gen species. Free Radic. Biol. Med. 68, 278–287 (2014)
18. Jayakumar, T.; Thomas, P.A.; Sheu, J.R.; Geraldine, P.: In-vitro
and in-vivo antioxidant effects of the oyster mushroom Pleurotus
ostreatus. Food Res. Int. 44, 851–861 (2011)
19. Figueiredo, L.; Régis, W.C.B.: Medicinal mushrooms in adjuvant
cancer therapies: an approach to anticancer effects and presumed
mechanisms of action. Nutrire 42, 1–10 (2017)
20. El-Deeb, N.M.; El-Adawi, H.I.; EL-wahab, A.E.A.; Haddad, A.M.;
El Enshasy, H.A.; He, Y.W.; Davis, K.R.: Modulation of NKG2D,
KIR2DL and cytokine production by Pleurotus ostreatus glucan
enhances natural killer cell cytotoxicity toward cancer cells. Front.
Cell Dev. Biol. 7, 165 (2019)
21. Pandey, A.T.; Pandey, I.; Hachenberger, Y.; Krause, B.C.; Haidar,
R.; Laux, P.; Singh, A.V.: Emerging paradigm against global
antimicrobial resistance via bioprospecting of mushroom into novel
nanotherapeutics development. Trends Food Sci. Technol. 106,
333–344 (2020)
22. Khan, M.A.; Tania, M.: Nutritional and medicinal importance of
Pleurotus mushrooms: an overview. Food Rev. Int. 28, 313–329
(2012)
23. Carrasco-González, J.A.; Serna-Saldívar, S.O.; Gutiérrez-Uribe,
J.A.: Nutritional composition and nutraceutical properties of the
Pleurotus fruiting bodies: potential use as food ingredient. J. Food
Compost. Anal. 58, 69–81 (2017)
24. Venturella, G.; Ferraro, V.; Cirlincione, F.; Gargano, M.L.: Medic-
inal mushrooms: bioactive compounds, use, and clinical trials. Int.
J. Mol. Sci. 22, 634 (2021)
25. Koutrotsios, G.; Tagkouli, D.; Bekiaris, G.; Kaliora, A.; Tsiaka, T.;
Tsiantas, K.; Chatzipavlidis, I.; Zoumpoulakis, P.; Kalogeropou-
los, N.; Zervakis, G.I.: Enhancing the nutritional and functional
properties of Pleurotus citrinopileatus mushrooms through the
exploitation of winery and olive mill wastes. Food Chem. 370,
131022 (2022)
26. Dkhil, M.A.; Diab, M.S.M.; Lokman, M.S.; El-Sayed, H.; Aljaw-
dah, H.M.A.; Al-Quraishy, S.; Bauomy, A.A.: Hepato-protective
effect of Pleurotus ostreatus extracts in cadmium-intoxicated rats.
J. King Saud Univ. Sci. 32, 3432–3436 (2020)
27. Goswami, B.; Majumdar, S.; Das, A.; Barui, A.; Bhowal, J.: Eval-
uation of bioactive properties of Pleurotus ostreatus mushroom
protein hydrolysate of different degree of hydrolysis. LWT 149(5),
111768 (2021)
28. Cateni, F.; Gargano, M.L.; Procida, G.; Venturella, G.; Cirlincione,
F.; Ferraro, V.: Mycochemicals in wild and cultivated mushrooms:
nutrition and health. Phytochem. Rev. 21, 339–383 (2022)
29. Pandey, A.T.; Pandey, I.; Kerkar, P.; Singh, M.P.: Antimicrobial
activity and mycochemical profile of methanol extract from Pleu-
rotus flabellatus.Vegetos34, 619–629 (2021)
30. Sánchez, C.: Bioactives from mushroom and their application. In:
Puri, M. (Ed.) Food Bioactives, pp. 23–57. Springer International
Publishing AG, Cham (2017)
31. Askarova, S.; Umbayev, B.; Masoud, A.; Kaiyrlykyzy, A.: The
links between the gut microbiome, aging, modern lifestyle and
Alzheimer’s disease. Front. Cell Infect. Microbiol. 10, 1–12 (2020)
32. Mwangi, R.W.; Macharia, J.M.; Wagara, I.N.; Bence, R.L.: The
antioxidant potential of different edible and medicinal mushrooms.
Biomed. Pharmacother. 147, 112621 (2022)
33. Croan, S.: Conversion of conifer waste into edible and medicinal
mushrooms. For. Prod. J. 54(2), 68–76 (2004)
34. Jonathan, S.; Okorie, A.; Garuba, E.; Babayemi, O.: Bioconversion
of sorghum stalk and rice straw into value added ruminant feed
using Pleurotus pulmonarius. Nat. Sci. 10(4), 10–16 (2012)
35. Maftoun, P.; Johari, H.; Soltani, M.; Malik, R.; Othman, N.Z.; El
Enshasy, H.A.: The edible mushroom Pleurotus spp.: I. Biodiver-
sity and nutritional values. Int. J. Biotechnol. Wellness Ind. 4(2),
67–83 (2015)
36. Smiderle, F.; Olsen, L.; Carbonero, E.; Baggio, C.; Freitas, C.; Mar-
con, R.; Santos, A.; Gorin, P.; Iacomini, M.: Anti-inflammatory and
analgesic properties in a rodent model of a (1→3), (1→6)-linked
β-glucan isolated fromPleurotus pulmonarius. Eur. J. Pharmacol.
597(1–3), 86–91 (2008)
37. Lin, P.; Yan, Z.F.; Kook, M.; Li, C.T.; Yi, T.H.: Genetic and chemi-
cal diversity of edible mushroom Pleurotus species. BioMed. Res.
Int. 2022, 1–13 (2022)
123

7282 Arabian Journal for Science and Engineering (2023) 48:7273–7283
38. Akyüz, M.; ¸Sule, ˙
I; Kırba˘g, S.: Nutrient content of Pleurotus pul-
monarius (Fr.) Quel. grown on some local lignocellulosic wastes.
KSUJ.Agric.Nat.25, 25–30 (2022)
39. Amirullah, N.A.; Abidin, N.Z.; Abdullah, N.; Manickam, S.: The
ultrasound extract of Pleurotus pulmonarius (Fr.) Quél alleviates
metabolic syndromes in hyperlipidaemic Wistar-Kyoto rats fed
with a high-fat diet. Biocatal. Agric. Biotechnol. 34, 102019 (2021)
40. Dawidowicz, L.: Cultivation of oyster mushrooms (Pleurotus spp.)
using organic waste: an example with Pleurotus pulmonarius (Fr.)
Quel. Folia. Biol. Oecol. 17, 104–110 (2021)
41. Banjo, N.; Abikoye, E.; Kukoye, A.: Comparison of three nutrient
supplements used as additive to sawdust during the cultivation of
oyster mushroom (Pleurotus pulmonarius). Niger. J. Microbiol. 18,
335–336 (2004)
42. Adebayo, G.; Omolara, B.; Toyin, A.: Evaluation of yield of oyster
mushroom (P. pulmonarius) grown on cotton waste and cassava
peel. Afr. J. Biotechnol. 8(2), 215–218 (2009)
43. Akinfemi, A.; Ogunwale, O.: Chemical composition and in vitro
digestibility of rice straw treated with Pleurotus ostreatus,Pleuro-
tus pulmonarius and Pleurotus tuber-regium.SlovakJ.Anim.Sci.
45(1), 14–20 (2012)
44. Zadrazil, F.: Cultivation of Pleurotus. In: Chang, S.T.; Hayes,
W.A. (Eds.) The Biology and Cultivation of Edible Mushrooms,
pp. 521–558. Academic Press, New York (1978)
45. Babu, D.R.; Pandey, M.; Rao, G.N.: Antioxidant and electrochem-
ical properties of cultivated Pleurotus spp. and their sporeless/low
sporing mutants. J. Food Sci. Technol. 51(11), 3317–3324 (2014)
46. Collins, C.M.; Lyne, P.M.: Microbiological methods. In: Collins,
C.H.,; Lyne, P.M.; Grange, J.M.; Flkinham, J.O. (Eds.) Mycologi-
cal Methods, p. 140, 8 ed. London (2004)
47. European Committee for Antimicrobial Susceptibility Testing
(EUCAST): Determination of minimum inhibitory concentrations
(MICs) of antibacterial agents by broth dilution. Clin. Microbiol.
Infect. Dis. 9, 1–7 (2003)
48. Erel, O.: A novel automated direct measurement method for total
antioxidant capacity using a new generation. More stable ABTS
radical cation. Clin. Biochem. 37, 277–285 (2004)
49. Erel, O.: A new automated colorimetric method for measuring total
oxidant status. Clin. Biochem. 38, 1103–1111 (2005)
50. Cuendet, M.; Hostettmann, K.; Potterat, O.; Dyatmiko, W.: Iridoid
glucosides with free radical scavenging properties from Fagraea
blumei. Helv. Chim. Acta. 80, 1144–1152 (1997)
51. Kirby, A.J.; Schmidt, R.J.: The antioxidant activity of Chinese
herbs for eczema and of placebo herbs. J. Ethnopharmacol. 56,
103–108 (1997)
52. Mossman, T.: Rapid colorimetric assay for cellular growth and
survival: application to proliferation and cytotoxicity assays. J.
Immunol. Methods. 65, 55–63 (1983)
53. Russo, A.; Acquaviva, R.; Campisi, A.; Sorrenti, V.; Di-Giacomo,
C.; Virgata, G.: Bioflavonoids as antiradicals. Antioxidants and
DNA cleavage protectors. Cell Biol. Toxicol. 16, 91–98 (2000)
54. Tepe, B.; Degerli, S.; Arslan, S.; Malatyali, E.; Sarikurkcu, C.:
Determination of chemical profile, antioxidant, DNA damage pro-
tection and antiamobic activities of Teucrium polium and Stachy
siberica. Fitoterapia 82, 237–246 (2011)
55. Chang, S.T.; Wasser, S.P.: The role of culinary-medicinal mush-
rooms on human welfare with a pyramid model for human health.
Int. J. Med. Mushrooms 14, 95–134 (2012)
56. Patel, S.; Goyal, A.: Recent developments in mushrooms as anti-
cancer therapeutics: a review. 3 Biotech. 2, 1–15 (2012)
57. Alves, M.; Ferreira, I.F.R.; Dias, J.; Teixeira, V.; Martins, A.;
Pintado, M.: A review on antimicrobial activity of mushroom
(basidiomycetes) extracts and isolated compounds. Planta Med.
78, 1707–1718 (2012)
58. Parihar, S.; Virani, K.D.; Pithawala, E.A.; Shukla, M.D.; Lahiri,
S.K.; Jain, N.K., et al.: Phytochemical screening, total phenolic
content, antibacterial and antioxidant activity of wild edible mush-
room Pleurotus ostreatus.Int.Res.J.Pharm.6, 65–69 (2015)
59. El-Batal, A.I.; Al-Hazmi, N.E.; Mosallam, F.M.; El-Sayyad, G.S.:
Biogenic synthesis of copper nanoparticles by natural polysaccha-
rides and Pleurotus ostreatus fermented fenugreek using gamma
rays with antioxidant and antimicrobial potential towards some
wound pathogens. Microb. Pathog. 118, 159–169 (2018)
60. Athukorala, Y.;Kim, K.N.; Jeon, Y.J.: Antiproliferative and antiox-
idant properties of an enzymatic hydrolysate from brown alg,
Ecklonia cava. Food Chem. Toxicol. 44, 1065–1074 (2006)
61. Kosani´c, M.; Rankovi´c, B.; Ranˇci´c, A.; Stanojkovi´c, T.: Evalu-
ation of metal concentration and antioxidant, antimicrobial, and
anticancer potentials of two edible mushrooms Lactarius delicio-
sus and Macrolepiota procera. J. Food Drug. Anal. 24, 477–484
(2016)
62. Shimada, K.; Fujikawa, K.; Yahara, K.; Nakamura, T.: Antioxida-
tive properties of xanthan on the autoxidation of soybean oil in
cyclodextrin emulsion. J. Agric. Food Chem. 40, 945–948 (1992)
63. Liu, X.; Zhou, B.; Lin, R.; Jia, L.; Deng, P.; Fan, K.; Zhang, J.:
Extraction and antioxidant activities of intracellular polysaccharide
from Pleurotus sp. mycelium. Int. J. Biol. Macromol. 47, 116–119
(2010)
64. Dulay, R.M.R.; Miranda, L.A.; Malasaga, J.S.; Kalaw, S.P.; Reyes,
R.G.; Hou, C.T.: Antioxidant and antibacterial activities of ace-
tonitrile and hexane extracts of Lentinus tigrinus and Pleurotus
djamour. Biocatal. Agric. Biotechnol. 9, 141–144 (2012)
65. Zhang, Z.; Zhang, Y.;Liu, H.; Wang, J.; Wang,D.; Deng, Z.; Zhong,
S.: A water-soluble selenium-enriched polysaccharide produced by
Pleurotus ostreatus: Purification, characterization, antioxidant and
antitumor activities in vitro. Int. J. Biol. Macromol. 168, 356–370
(2021)
66. Sudha, G.; Vadivukkarasi, S.; Shree, R.B.I.; Lakshmanan, P.:
Antioxidant activity of various extracts from an edible mushroom
Pleurotus eous. Food Sci. Biotechnol. 21, 661–668 (2012)
67. Song, X.; Ren, Z.; Zhang, C.: Antioxidant, anti-inflammatory
and renoprotective effects of acidic-hydrolytic polysaccharides by
spent mushroom compost (Lentinula edodes) on LPS-induced kid-
ney injury. Int. J. Biol. Macromol. 151, 1267–1276 (2019)
68. Kim, M.Y.; Seguin, P.; Ahn, J.K.; Kim, J.J.; Chun, S.C.; Kim, E.H.;
Seo, S.H.; Kang, E.Y.; Kim, S.L.; Park, Y.J.; Ro, H.M.; Chung,
I.M.: Phenolic compound concentration and antioxidant activities
of edible and medicinal mushrooms from Korea. J. Agric. Food
Chem. 56, 7265–7270 (2008)
69. Finimundy, T.C.; Gambato, G.; Fontana, R.; Camassola, M.; Sal-
vador, M.; Moura, S.; Hess, J.; Henriques, J.A.P.; Dillon, A.J.P.;
Roesch-Ely, M.: Aqueous extracts of Lentinula edodes and Pleu-
rotus sajor-caju exhibit high antioxidant capability and promising
in vitro antitumor activity. Nutr. Res. 33, 76–84 (2013)
70. G˛asecka, M.; Mleczek, M.; Siwulski, M.; Niedzielski, P.: Phenolic
composition and antioxidant properties of Pleurotus ostreatus and
Pleurotus eryngii enriched with selenium and zinc. Eur. Food Res.
Technol. 242, 723–732 (2016)
71. Vieira, P.A.F.; Gontijo, D.C.; Vieira, B.C.; Fontes, E.A.; de
Assunção, L.S.; Leite, J.P.V.; Kasuya, M.C.M.: Antioxidant activ-
ities, total phenolics and metal contents in Pleurotus ostreatus
mushrooms enriched with iron, zinc or lithium. LWT 54, 421–425
(2013)
72. Song, F.Q.; Liu, Y.; Kong, X.S.; Chang, W.; Song, G.: Progress
on understanding the anticancer mechanisms of medicinal mush-
room: Inonotus obliquus. Asian Pac. J. Cancer Prev. 14, 1571–1578
(2013)
73. Ma, G.; Yang, W.; Fang, Y.; Ma, N.; Pei, F.; Zhao, L.; Hu, Q.:
Antioxidant and cytotoxicites of Pleurotus eryngii residue polysac-
charides obtained by ultrafiltration. LWT 73, 108–116 (2016)
74. Khan, A.A.; Gani, A.; Ahmad, M.; Masoodi, F.A.; Amin, F.;
Kousar, S.: Mushroom varieties found in the Himalayan regions
123

Arabian Journal for Science and Engineering (2023) 48:7273–7283 7283
of India: antioxidant, antimicrobial, and antiproliferative activities.
Food Sci. Biotechnol. 25, 1095–1100 (2016)
75. Sun, H.; Zhu, Z.; Tang, Y.; Ren, Y.; Song, Q.; Tang, Y.; Zhang,
Y.: Structural characterization and antitumor activity of a novel
Se-polysaccharide from seleniumenriched Cordyceps gunnii. Food
Funct. 9, 2744–2754 (2018)
76. Maity, G.N.; Maity, P.; Choudhuri, I.; Bhattacharyya, N.; Acharya,
K.; Dalai, S.; Mondal, S.: Structural studies of a water insoluble β-
glucan from Pleurotus djamor and its cytotoxic effect against PA1,
ovarian carcinoma cells. Carbohydr. Polym. 222, 114990 (2019)
77. Elhusseiny, S.M.; El-Mahdy, T.S.; Awad, M.F.; Elleboudy, N.S.;
Farag, M.; Aboshanab, K.M.; Yassien, M.A.: Antiviral, cytotoxic,
and antioxidant activities of three edible agaricomycetes mush-
rooms: Pleurotus columbinus,Pleurotus sajor-caju,andAgaricus
bisporus. J. Fungi 7, 645 (2021)
78. Kim, K.C.; Kim, I.G.: Ganoderma lucidum extract protects DNA
from strand breakage caused by hydroxyl radical and UV irradia-
tion. Int. J. Mol. Med. 4, 273–280 (1999)
79. Sa-Ard, P.; Sarnthima, R.; Khammuang, S.; Kanchanarach, W.:
Antioxidant, antibacterial and DNA protective activities of pro-
tein extracts from Ganoderma lucidum. J. Food Sci. Technol. 52,
2966–2973 (2015)
80. Akyüz, M.; Karagöz, I.D.; ˙
Inci, ¸S; Kılıç, H.˙
I; Kırba˘g, S.; Sim-
itçio˘glu, B.: Cytotoxicity and DNA protective effects of Terf a z ia
and Picoa species from eastern region of Turkey. Istanbul J. Pharm.
51, 198–203 (2021)
Springer Nature or its licensor (e.g. a society or other partner) holds
exclusive rights to this article under a publishing agreement with the
author(s) or other rightsholder(s); author self-archiving of the accepted
manuscript version of this article is solely governed by the terms of such
publishing agreement and applicable law.
123