Antioxidant and Radical Scavenging Activity of Nine Edible Mushrooms Extract
This study revealed that antioxidant properties, 1, 1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging activity and reducing power were observed in different mushroom species namely Agaricus bisporus, Pleurotus sajor-caju, Pleurotus euos, Hybsizus ulmarius, Pleurotus florida, Volveriela volvaciea, Pleurotus platypus, Pleurotus djamor and Calocybe indica. Methonolic extract of P. euos was recorded maximum levels of DPPH free radical scavenging (6 mg mL-1), reducing power (6 mg mL-1) and enzymatic antioxidants viz., catalase, superoxide dismutase and peroxidise, 68.7±1.1, 0.39±0.01, 42.21±0.06, 37.12±0.04 and 7.21±0.05, respectively. The maximum amount of enzymatic antioxidants were recorded viz., CAT, SOD and POX; 42.21±0.06, 37.12±0.04 and 7.21±0.05 μmol, respectively.
Free radicals are responsible for aging and causing various human diseases.
A study shows that antioxidant substances which scavenge free radicals play
an important role in the prevention of free radical-induced diseases. By donating
hydrogen radicals, the primary radicals are reduced to non radical chemical
compounds and are then converted to oxidize antioxidant radicals (Jadhav
et al., 1995; El-Enshasy et al., 2010;
Yamagushi et al., 1998). This action helps in
protecting the body from degenerative diseases. Epidemiological studies have
shown the beneficial effects of diets rich in vegetables, fruits and grain products
in reducing the risk of cardiovascular disease and certain cancers (Beecher,
1999). The degenerative diseases associated with aging include cancer, cardiovascular
disease, immune-system decline, brain dysfunction and cataracts (Ames
et al., 1993). Weisburger (1999) and Ames
et al. (1993) reported that, the consumption of plant foods, such
as fruits, vegetables, red wines and juices provides protection against various
diseases, including cancer, cardio and cerebrovascular diseases. Exogenous chemical
and endogenous metabolic processes in the food system might produce highly reactive
free radicals, especially oxygen derived radicals, which are capable of resulting
in cell death and tissue damage. However, antioxidant supplements containing
foods might be used to help human body cells to reduce oxidative scratch (Halliwell
and Gutteridge, 1999). Mushrooms are considered to be a good source of antioxidants,
such as variegatic acid and diboviquinone, which have been found in mushrooms
(Kasuga et al., 1995). Cheung
et al. (2003) reported that the methanol and water extract mushrooms
were found to rich antioxidative activities. Mushroom species had been shown
to possess antioxidant capacity in in vitro systems (Ribeiro
et al., 2006). Seline and Johein (2007) reported
that the l-carnitine concentration in mushroom ranged from 133 to 530 mg kg-1
DM (mean 320 mg kg-1 DM). The aim of this study was to determine
the antioxidant property, scavenging activity and reducing power in the fruiting
bodies of 9 edible mushroom species were collected from Namakkal district, Tamil
Nadu and Southeast India.
MATERIALS AND METHODS
Mushroom collection: The species were collected from Namakkal district,
Tamil Nadu and Southeast India, during April to June in 2009. The study was
carried out of Centre of Advanced study in Marine Biology, Annamalai University,
Parngipettai, Tamil Nadu, India.
Estimation of antioxidant activity: A fine dried mycelial mat (biomass
filtrate paper) and harvested mushroom powder (20 meshes) samples (5 g) were
continuously extracted with methanol in a Soxhlet apparatus for 24 h. The methanolic
extract was evaporated to dryness at 40°C and redissolved in methanol at
a concentration of 5 mg mL-1 and stored at 4°C prior to further
use (Barros et al., 2007).
CAT: The mean CAT activity (μmol of H2O2
consumed/min/mg protein) was assayed following the method described by Sinha
SOD: The mean activity of SOD (units/min/mg protein) was determined
by the method of Marklund and Marklund (1974), in which
one unit was considered to be the amount of enzyme that inhibited pyrogallol
autooxidation by 50%.
POX: Mean POX activity (μmol of pyrogallol oxidized/min/mg protein)
of the mushroom extracts was measured according to the method described by Sadasivam
and Manickam (2004).
DPPH radical scavenging activity: The scavenging activity of the free
and bound extracts on 1, 1-diphenyl 2-picrylhydrazyl (DPPH) radical was measured
according to the method of Cheung et al. (2003)
with some modifications. Aliquots (various concentration of methanol extract)
of 0.8 mL of 0.2 mM DPPH methanolic solution was mixed with 0.2 mL of the extracts.
The mixture was vigorously shaken and left to stand for 10 min under subdued
light. The absorbance was measured at 520 nm. The DPPH radical scavenging activity
(%) was calculated by the following equation:
where, Asample is the absorbance in the presence of sample and Acontrol is the absorbance in the absence of sample, respectively. All extracts were analyzed in triplicate.
Reducing power: The reducing power was determined according to the method
of Oyaizu (1986) with some modifications. Various concentrations
of methanolic extracts from mushrooms (2.5 mL) were mixed with 2.5 mL of 200
mM sodium phosphate buffer (pH 6.6, Wako Pure Chemical Co., Osaka, Japan) and
2.5 mL of 1% potassium ferricyanide (Sigma) and the mixture was incubated at
50°C for 20 min. After 2.5 mL of 10% trichloroacetic acid (w/v, Wako) were
added, the mixture was centrifuged at 650 rpm for 10 min. The upper layer (5
mL) was mixed with 5 mL of deionised water and 1 mL of 0.1% ferric chloride
(Wako) and the absorbance was measured at 700 nm in a 2020 double beem spectrophotometer.
A higher absorbance indicates a higher reducing power.
Statistical analysis: All experiments were done in three replicates and mean values are presented. Statistical analysis was performed on the data by Duncans multiple range test with means followed by a common letter are not significantly different at the 5% level by DMRT. Values are expressed as Mean±SD.
RESULTS AND DISCUSSION
Antioxidant, DPPH and reducing power results were clearly indicated that among
sp., P. eous was significantly increased the enzymatic antioxidative
substances, compared to other sp. The maximum amount of enzymatic antioxidants
were recorded viz., CAT, SOD and POX; 42.21±0.06, 37.12±0.04 and
7.21±0.05 μmol, respectively (Table 1). Yang
et al. (2002) was reported methanol extraction with; two mushrooms
strains had the highest antioxidant activity. This report was similar values
to compare our study. In the past few years, the suspected toxicity of some
synthetic compounds used in food has raised interest in natural products (Stone
et al., 2003). Some food industries and pharmaceuticals have increased
their efforts in preparing bioactive compounds from natural sources by extraction
and purification. Antioxidant compounds can scavenge free radicals and increase
shelf life by retarding the process of lipid peroxidation, which is one of the
major reasons for deterioration of food products during processing and storage
(Halliwell and Gutteridge, 1999). So we need for identifying
alternative natural and secure sources of food antioxidants has been created
and the search for natural antioxidants, particularly of plant origin, has remarkably
increased in recent years (Skerget et al., 2005).
Jose and Janardhanan (2000) have reported that the
methanol extracts of Pleurotus sp., exhibited effective radical scavenging.
The concentrations of cysteine, methionine and aspartic acid are reported to
be higher in P. ostreatus than those in other edible mushrooms, such
as Agaricus sp. and Lentinula sp. (Mattila
et al., 2002). In addition, P. ostreatus has also been reported
to possess excellent reducing power of ferric ions (Lin, 1999).
In this study, methanolic extracts were prepared in different concentration;
these are 2, 4 and 6 mg mL-1. DPPH scavenging activities and reducing
power in various concentrations of methanol extracts were also higher in P.
euos; this was followed by A. bisporus (Fig. 1,
||Antioxidant activities of catalase, superoxide dismutase and
glutathione peroxidase in dried mushroom
|Values are expressed as Mean±SD of nine mushroom sp.
in each group. Mean of three replications. Means followed by a common letter
are not significantly different at the 5% level by DMRT. Units: CAT: 1 μmole
of H2O2 utilized /min /mg protein, SOD: μmole
of Inhibition of 50% nitrite formation/min/mg protein, POX: 1 μmole
of pyrogallol oxidized/min/mg protein
||Scavenging effect of methanolic extracts from 9 edible mushrooms
on 1, 1-diphenyl-2-picrylhydrazyl radical. Each value is expressed as Mean±SD.
spp1: Agaricus bisporus, spp2: Pleurotus sajor-caju, spp3:
Pleurotus euos, spp4: Hybsizus ulmarius, spp5: Pleurotus
florida, spp6: Volveriela volvaciea, spp7: Pleurotus platypus,
spp8: Pleurotus djamor and spp9: Calocybe indica
||Reducing power of different concentrations of methanol extract
of 9 edible mushrooms. Values are expressed as Mean±SD. spp1: Agaricus
bisporus, spp2: Pleurotus sajor-caju, spp3: Pleurotus euos,
spp4: Hybsizus ulmarius, spp5: Pleurotus florida, spp6: Volveriela
volvaciea, spp7: Pleurotus platypus, spp8: Pleurotus djamor
and spp9: Calocybe indica
Different concentrations of DPPH scavenging activities of methanolic extracts
from P. eous were 34.7 to 68.5%. The methanolic extract of mushroom species
has presented in significant antioxidant activity. Antioxidant activity of the
mushroom extracts include scavenging of free radicals also find good accountable.
In addition, the mushroom is utilize, the source of natural antioxidants in
We would like to thank Dean and the authorities of the Annamalai University, Parangipettai for the facilities provided. We also thank the anonymous reviewers and editor for helpful comments that improved the manuscript.
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