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OPEN ACCESS EC NUTRITIONEC NUTRITION
Research Article
Antioxidant and Ergothioneine Assessment of Nutritionally
Enriched Agaricus bisporus Powders
Supriya Yadav* and Jude Wilson
MBio, Monaghan Mushrooms Ireland Unlimited Company, Tyholland, Ireland
Citation: Supriya Yadav and Jude Wilson. “Antioxidant and Ergothioneine Assessment of Nutritionally Enriched Agaricus bisporus
Powders”. EC Nutrition 17.2 (2022).
*Corresponding Author: Supriya Yadav, MBio, Monaghan Mushrooms Ireland Unlimited Company, Tyholland, Ireland.
Received: November 22, 2021
Abstract
MBio produces a range of nutritionally enriched Agaricus bisporus- derived powder products, such as vitamin D enriched mush-
room powders, selenium enriched mushroom powders, and vitamin B12 enriched mushroom powders that may have numerous
mushroom powders and a whole mushroom powder. This study reports comparative antioxidant assessment of MBio’s proprietary
Agaricus bisporus
was evaluated through in vitro free radical scavenging activity (DPPH, nitric oxide, ABTS and hydroxyl radical), total polyphenols
(5.18 - 9.23 mg/g) and polyphenol content (5.66 - 5.94 mg/g) followed by the vitamin D enriched mushroom powder (DPPH 3.33
- 5.86 mg/g, polyphenols 4.86 - 5.73 mg/g), the selenium enriched mushroom powder (DPPH 2.57 - 5.05 mg/g, polyphenols 4.32 -
5.58 mg/g) and the whole mushroom powder (DPPH 3.36 - 6.72 mg/g, polyphenols 4.56 - 5.52 mg/g) compared to the commercial
mushroom powders. Furthermore, the selenium enriched mushroom powder exhibited the highest nitric oxide scavenging activity
(NOSA 23.18 - 25.73 mg/g) and the lowest IC50 (1.35 - 2.89 mg) for scavenging the ABTS radical indicating better antioxidant activity
-
power (1303 mg/kg) and the selenium enriched mushroom powder (1290.2 mg/Kg) exhibited higher ergothioneine compared to the
-
Agaricus bisporus powder products can
Keywords: Antioxidant; Agaricus bisporus; Mushroom Powders; Nutritionally Enriched; Selenium; Ergothioneine
Abbreviations
OS: Oxidative Stress; ROS: Reactive Oxygen Species; WMP: Whole Mushroom Powder; DPPH: 2,2-Diphenyl-1-Picrylhydrazyl Reagent;
NOSA: Nitric Oxide Radical Scavenging Activity; OHSA: Hydroxyl Radical Scavenging Activity; HPLC: High-Performance Liquid Chromatog-
Citation: Supriya Yadav and Jude Wilson. “Antioxidant and Ergothioneine Assessment of Nutritionally Enriched Agaricus bisporus
Powders”. EC Nutrition 17.2 (2022).
Antioxidant and Ergothioneine Assessment of Nutritionally Enriched Agaricus bisporus Powders
Introduction
The ROS have adverse reactions with polyunsaturated fatty acids, proteins and nucleotides that could lead to lipid peroxidation, inacti-
vated proteins and impaired DNA and RNA. If there is no adequate defence against ROS by enzymatic and non-enzymatic antioxidants, it
damage leading to a range of disorders such as aging, Alzheimer’s disease, Parkinson’s disease, immunological and many other neural
Among all free radical species, hydroxyl and superoxide radicals are found to be the main culprits in the damage that free radicals in-
the brain, vascular endothelial cells and phagocytes converts L-arginine to nitric oxide radical. Nitric oxide (NO) radical has an unpaired
electron, which reacts with certain types of proteins and other free radicals. The toxicity of NO becomes adverse when it reacts with
superoxide radical, forming a highly reactive peroxynitrite anion (ONOO
In normal body conditions, the balance between free radicals is maintained by the body’s antioxidant system. This is encompassed by
enzymes, such as superoxide dismutase and catalase, and/or by compounds, such as ascorbic acid (vitamin C), tocopherols and glutathi-
one. However, in certain situations, this mechanism of antioxidant protection becomes unbalanced and, therefore, antioxidants sourced
through the diet play important role in reducing oxidative damage.
-
rooms are an excellent source of nutrients, including B vitamins and minerals such as selenium, copper and potassium. Mushrooms are
-
-
been found to contain high levels of antioxidants, including phenolic compounds and, in particular, the sulphur-containing amino acid
A. bisporus, such as terpenes, diterpenes, indoles and
Ergothioneine is mainly biosynthesized in fungi, some cyanobacteria and mycobacteria using histidine with cysteine and methionine,
exhibited an antioxidant role and was proposed as a possible therapeutic for prevention of erythrocytes disorders due to oxidative dam-
MBio produces a range of nutritionally enriched Agaricus bisporus
Citation: Supriya Yadav and Jude Wilson. “Antioxidant and Ergothioneine Assessment of Nutritionally Enriched Agaricus bisporus
Powders”. EC Nutrition 17.2 (2022).
Antioxidant and Ergothioneine Assessment of Nutritionally Enriched Agaricus bisporus Powders
Aim of the Study
The current study was aimed to assess the mushroom powders using various important radical scavenging activities including DPPH,
nitric oxide, hydroxyl and ABTS radical, total polyphenol and ergothioneine content.
Materials and Methods
Chemicals
DPPH (2,2-diphenyl-1-picrylhydrazyl) reagent, methanol, trolox (6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid) as a stan-
dard antioxidant, for polyphenols: Folin Ciocalteau reagent (2N), sodium bicarbonate (NaHCO3), for NOSA: sodium nitroprusside (SNP),
phosphate buffered saline (PBS pH 7.4), sulphanilamide, glacial acetic acid, naphthylethylene diamine hydrochloride (NED), for ABTS:
ABTS reagent (Sigma), manganese dioxide (MnO2), for OHSA: ascorbic acid, ferrous chloride (FeCl3), H2O2, EDTA, 2deoxy ribose, thio-
barbituric acid (TBA), trichloroacetic acid (TCA). Trolox and gallic acid were used as a standard antioxidant. Ergothioneine analysis was
conducted at CampdenBRI group UK on contractual basis. All the materials and chemicals were obtained from Lennox Ireland and Sigma.
Preparation of mushroom extracts for antioxidant analysis
A number of MBio’s proprietary mushroom powder products (Table 1) were obtained from MBio, Monaghan Mushrooms Ireland
Unlimited Company. The random commercial market mushroom powder samples were used as commercial powders for antioxidant
comparison. The details of the production of MBio’s mushroom powder products, source and nutritional composition is described below.
Each sample (0.1g) was extracted in 10 mL of the extracting solvent (Table 2) for 14h at room temperature in a shaking incubator. The
Mushroom Powders Codes
Whole mushroom powder (WMP) Batch 1
Batch 2
Batch 3
Batch 1
Batch 2
Batch 3
Batch 1
Batch 2
Batch 3
Selenium enriched mushroom powder (5 - 10 mg/kg) Batch 1
Batch 2
Batch 3
Commercial Powder of Shitake mushrooms CP1
Commercial Powder of Chaga mushrooms CP2
Commercial Powder of Reishi mushrooms CP3
Commercial Powder of Lion’s Mane mushrooms CP4
Commercial Powder of a mixture of different mushrooms CP5
Table 1:
Citation: Supriya Yadav and Jude Wilson. “Antioxidant and Ergothioneine Assessment of Nutritionally Enriched Agaricus bisporus
Powders”. EC Nutrition 17.2 (2022).
Antioxidant and Ergothioneine Assessment of Nutritionally Enriched Agaricus bisporus Powders
Vitamin D enriched mushroom powder
The source of vitamin D2 mushroom powder is commercially cultivated, freshly harvested fruiting bodies and stalk component of
Agaricus bisporus obtained from single batch, which is sourced from Monaghan Mushrooms Ireland Unlimited Company. The vitamin D2
-
ferences in the nutritional information for fresh Agaricus bisporus and novel vitamin D enriched mushrooms bar a concentration effect
intake for adults for copper, potassium, selenium and vitamin D. In addition, the vitamin D2 mushroom powder is also high in proteins,
Vitamin B12 enriched mushroom powder
The vitamin B12 enriched mushroom powder is derived from freshly harvested Agaricus bisporus, common button mushrooms natu-
rally enriched with B12 during the cropping cycle, obtained from single batch, dried and milled under mild conditions to preserve its
sustainably sourced, fully traceable and derived from a vegan source.
Selenium enriched mushroom powder
The selenium enriched mushroom powder is derived from freshly harvested Agaricus bisporus, common button mushrooms naturally
enriched with selenium during the cropping cycle, obtained from single batch, dried and milled under mild conditions to preserve its
-
tainably sourced, fully traceable and derived from a vegan source.
Whole mushroom powders
The whole mushroom powder is derived from freshly harvested fruiting bodies and stalk component of Agaricus bisporus including
white closed cup mushrooms obtained from single batch, dried and milled under mild conditions to preserve its nutrient rich content.
derived from a vegan source.
The powders were analysed for nutritional composition, which revealed that the mild drying process does not affect the nutritional
status of the powders, bar a concentration effect from the removal of water. Furthermore, whole mushroom powder nutritional analysis
and also high in minerals, which further contributes to the nutritional value of the Agaricus bisporus.
Assays Extraction Solvents Standards Wavelengths (nm)
DPPH Methanol Trolox 515
Total poly-
phenols
Water Gallic acid 700
NOSA Water Gallic acid 540
ABTS Methanol Trolox 734
OHSA Water Gallic acid 520
Table 2: Antioxidant assay extraction solvents and standards.
Citation: Supriya Yadav and Jude Wilson. “Antioxidant and Ergothioneine Assessment of Nutritionally Enriched Agaricus bisporus
Powders”. EC Nutrition 17.2 (2022).
Antioxidant and Ergothioneine Assessment of Nutritionally Enriched Agaricus bisporus Powders
Commercial mushroom powders
Randomly available marketed mushroom powders were obtained from online shops. The samples include different strains of mush-
room powder such as: Shitakeii, Chaga, Reishi, Lion’s Mane and commercial mixture of different mushroom powders. During the experi-
ments, the marketed mushroom powders were coded as commercial powders starting from CP1, CP2, CP3, CP4 and CP5, respectively.
DPPH radical scavenging activity
DPPH is a well-known radical that scavenges other radicals, which is used as an indicator of the rate reduction of a chemical reaction.
DPPH radical shows deep violet colour in solution, which upon neutralization, it changes to pale yellow or colourless. Therefore, the reac-
of the absorbance at 515 nm was monitored and expressed as mg of trolox equivalent DPPH radical scavenging activity. The samples were
analysed in triplicate.
Total polyphenol content
A polyphenol contains polyphenolic substructure, which acts as an antioxidant by scavenging the free radicals and up-regulates certain
-
distilled water followed by 0.2 mL of Folin Ciocalteau reagent (1:1 diluted reagent with distilled water (D/W)) except sample blanks. After
3 in D/W was added and the volumes were made up to 3 mL with D/W. The samples were
results were expressed as mg of gallic acid equivalents (GAE)/g of the sample. Analysis was completed in triplicate.
Nitric oxide scavenging activity (NOSA)
The procedure was based on the principle that sodium nitroprusside (SNP) in aqueous solution at physiological pH spontaneously
coupling with NED was measured spectrophotometrically at 540 nm against a blank sample. Gallic acid (0.1 mg/mL) was used as a stan-
dard. All tests were performed in triplicate.
ABTS radical scavenging activity
Free radicals are an outcome of various metabolic activities and their excess production leads to many diseases. According to Scopus
citation, the 2,2 -azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS•+) radical cation-based assays and the 2,2-diphenyl-1- picryl-
hydrazyl (DPPH) radical-based assays are most widely used to predict the antioxidant activity. The trolox equivalent antioxidant activity
(TEAC) was estimated using the ABTS•+et al
was evaluated against ABTS•+ radical, which was generated by oxidizing a 5 mM of ABTS (2, 2’-azinobis 3-ethylbenzothiazoline-6-sulfonic
Citation: Supriya Yadav and Jude Wilson. “Antioxidant and Ergothioneine Assessment of Nutritionally Enriched Agaricus bisporus
Powders”. EC Nutrition 17.2 (2022).
Antioxidant and Ergothioneine Assessment of Nutritionally Enriched Agaricus bisporus Powders
acid) diammonium salt, with manganese dioxide in PBS (pH 7.4) at ambient temperature for 14h in dark. The reaction mixture contained
2.0 mL of ABTS•+
the standard trolox (0.1 mg/mL in methanol) as a positive control. The results were expressed as IC50 as mg of the powder required for
•+ radical. Each sample was analysed in triplicate for IC50. Lower the IC50 is considered as better antioxidant activity
equivalent to Trolox.
Hydroxyl radical scavenging activity (OHSA)
The hydroxyl radical is one of the potent reactive oxygen species that reacts with polyunsaturated fatty acid moieties of cell membrane
-
cluding atherosclerosis, cancer and neurological disorders, and can be prevented by the action of non-reducing substances. OHS assay was
conducted using ascorbic acid-iron-EDTA model of hydroxyl radical generating system. The total volume of the reaction mixture was 1 mL,
which included 0.4 mL sodium phosphate buffer (20 mM, pH7.4), 0.1 mL of the sample extract, 0.1 mL of 2-deoxy ribose (60 nM/L), 0.1
mL ferric chloride (1mM/L), 0.1 mL EDTA (1.04 mM/L), 0.1 mL ascorbic acid (2 mM/L) and the reaction was initiated by adding 0.1 mL of
H2O2 (10 mM/L). The standard gallic acid (0.1 mg/mL) was used ranging from 0.02 - 0.1 mL for the standard curve. The reaction mixture
measured at 532 nm. The sample blanks were prepared by replacing TBA with 1 mL of water. The results were expressed as mg of gallic
acid equivalent/g of sample. Analysis was conducted in triplicate.
Ergothioneine analysis
-
used for detection. The mass spectrophotometry was run in positive ionisation mode optimized for the detection of ergothioneine. One
Statistical analysis
The data was analysed using MS-EXCEL. One way Anova was used for statistical analysis for comparison across batches, different type
of mushroom powders.
Results
DPPH radical scavenging activity
Whole mushroom powder: The DPPH scavenging activity was reported as equivalent to standard antioxidant Trolox. The DPPH scaveng-
Citation: Supriya Yadav and Jude Wilson. “Antioxidant and Ergothioneine Assessment of Nutritionally Enriched Agaricus bisporus
Powders”. EC Nutrition 17.2 (2022).
Antioxidant and Ergothioneine Assessment of Nutritionally Enriched Agaricus bisporus Powders
Average ± SD
Mushroom
powders
DPPH (mg
equivalent
to trolox/g
powder)
Total Polyphenols
(mg equivalent
to gallic acid/g
powder)
NOSA (mg
equivalent to
gallic acid/g
powder)
ABTS (IC50:mg of
powder equivalent
to trolox for 50%
inhibition of ABTS)
OHSA (mg
equivalent to
gallic acid/g
powder)
Ergothioneine
(mg/Kg
powder)
WMP 1 4.72 ± 0.15 5.52 ± 0.31 15.35 ± 3.82 2.81 ± 0.41 26.23 ± 2.81 547
2 6.72 ± 1.70 4.82 ± 0.37 14.00 ± 1.32 1.87 ± 0.37 20.78 ± 1.28 1009
3 3.36 ± 1.26 4.56 ± 0.18 14.54 ± 1.03 2.67 ± 0.42 29.96 ± 1.63 1009
1 5.86 ± 1.53 4.86 ± 0.27 14.22 ± 2.17 1.93 ± 0.37 24.38 ± 2.08 978.5
2 3.33 ± 1.28 5.05 ± 0.25 15.71 ± 1.41 2.99 ± 0.42 24.98 ± 5.24 1087
3 5.03 ± 0.40 5.73 ± 1.07 11.63 ± 0.33 2.46 ± 0.17 10.87 ± 1.16 1843.5
1 9.23 ± 2.35 5.91 ± 0.49 14.32 ± 1.39 1.62 ± 0.32 38.37 ± 4.38 541.5
2 5.18 ± 1.30 5.94 ± 0.19 21.88 ± 0.09 1.74 ± 0.19 17.39 ± 2.37 189.5
3 5.31 ± 0.17 5.66 ± 0.18 14.75 ± 3.21 2.89 ± 0.62 24.98 ± 1.36 827
Selenium 1 5.05 ± 0.43 5.58 ± 0.08 25.73 ± 1.93 1.13 ± 0.16 25.59 ± 0.13 1266
2 3.07 ± 1.08 4.59 ± 0.03 23.35 ± 0.66 1.35 ± 0.21 26.98 ± 0.99 1160.5
3 2.57 ± 0.88 4.31 ± 0.06 23.18 ± 0.83 2.09 ± 0.18 23.98 ± 0.44 1444
Com-
mercial
powders
CP1 1.19 ± 0.21 3.43 ± 0.58 11.35 ± 2.85 6.82 ± 1.83 9.498 ± 0.78 95.5
CP2 0.96 ± 0.62 5.43 ± 1.14 14.91 ± 1.77 8.62 ± 0.82 12.34 ± 1.27 1408
CP3 3.05 ± 0.90 3.24 ± 0.45 13.05 ± 1.27 4.48 ± 0.17 17.09 ± 0.87 166
CP4 0.40 ± 0.30 0.62 ± 0.15 9.70 ± 2.60 11.71 ± 0.97 12.18 ± 2.06 20
CP5 0.76 ± 0.16 1.04 ± 0.19 11.17 ± 1.77 9.93 ± 1.44 11.06 ± 0.93 35
Table 3: Comparison of antioxidant activity different mushroom powders.
Figure 1: DPPH scavenging activities of mushroom powders.
Citation: Supriya Yadav and Jude Wilson. “Antioxidant and Ergothioneine Assessment of Nutritionally Enriched Agaricus bisporus
Powders”. EC Nutrition 17.2 (2022).
Antioxidant and Ergothioneine Assessment of Nutritionally Enriched Agaricus bisporus Powders
Vitamin D enriched mushroom powder
-
Vitamin B12 enriched mushroom powder: The DPPH scavenging activity ranged from 5.18-9.23 mg/g. Batch 1 (9.23 mg/g) exhibited
and table 3), which might be dependent on the initial concentration of the fresh B12- enriched mushrooms used for the production of the
vitamin B12 enriched mushroom powder.
Selenium enriched mushroom powder: Three independent batches were assessed for the DPPH activity. Batch 2 (3.07 mg/g) and Batch
mushroom powder (2.57 - 5.05 mg/g) was observed, which might be dependent on the variability of the original selenium concentration
of the fresh selenium- enriched mushrooms.
Among nutritionally enriched mushroom powders, the vitamin B12 enriched mushroom powder exhibited higher DPPH activity (Fig-
Agaricus bisporus derived mushroom pow-
Comparison of nutritionally enriched mushroom powders vs marketed available mushroom powders: The results from com-
marketed powders (CP1 (1.19mg/g), CP2 (0.96 mg/g), CP4 (0.40 mg/g) and CP5 (0.76mg/g)) (Figure 1). CP3 (3.05 mg/g) exhibited no
powders exhibited strong antioxidant activity compared to the commercial powders.
Total polyphenolic content
Whole mushroom powder
Vitamin D enriched mushroom powder
Overall, the vitamin D enriched mushroom powders exhibited strong retention of polyphenol content across the batches (Figure 2 and
table 3).
Vitamin B12 enriched mushroom powder
-
mushroom powder indicated consistent retention of polyphenol content (Figure 2 and table 3).
Selenium enriched mushroom powder
Citation: Supriya Yadav and Jude Wilson. “Antioxidant and Ergothioneine Assessment of Nutritionally Enriched Agaricus bisporus
Powders”. EC Nutrition 17.2 (2022).
Antioxidant and Ergothioneine Assessment of Nutritionally Enriched Agaricus bisporus Powders
content. Although, the three different batches exhibited variation for polyphenol content, this might be due to the initial concentration of
selenium in the fresh selenium enriched mushrooms used for development of the selenium enriched mushroom powders (Figure 2 and
table 3).
-
of WMP and the selenium enriched mushroom powder (Figure 2).
Figure 2: Total polyphenol content of mushroom powders.
Comparison of nutritionally enriched mushroom powders vs marketed available mushroom powders
-
room powders such as CP1, CP3, CP4 and CP5 mushroom powders (3.43, 3.24, 0.62 and 1.04 mg/g, respectively). Among commercial
polyphenol content compared to the marketed mushroom powders.
Nitric oxide radical scavenging activity (NOSA)
Whole mushroom powder
-
oxidant activity among all batches of WMP (Figure 3 and table 3).
Vitamin D enriched mushroom powder
-
retention of the NOSA (Figure 3 and table 3) indicating strong antioxidant activity.
Citation: Supriya Yadav and Jude Wilson. “Antioxidant and Ergothioneine Assessment of Nutritionally Enriched Agaricus bisporus
Powders”. EC Nutrition 17.2 (2022).
Antioxidant and Ergothioneine Assessment of Nutritionally Enriched Agaricus bisporus Powders
Vitamin B12 enriched mushroom powder
-
min B12 concentration in the vitamin B12 enriched mushrooms used for development of the vitamin B12 enriched mushroom powders.
Selenium enriched mushroom powder: All batches of the selenium enriched mushroom powder (25.73, 23.35, 23.18 mg/g, respec-
of nitric oxide scavenging activity among batches of the selenium enriched mushroom powders (Figure 3 and table 3).
Comparison of nutritionally enriched mushroom powders vs marketed available mushroom powders: Finally, in line with above
-
room powders (CP1 (11.35mg/g), CP2 (14.91 mg/g), CP3 (13.05 mg/g), CP4 (9.70 mg/g) and CP5 (11.17 mg/g) powders). Although, CP2
and CP3 powders exhibited NOSA equivalent to the vitamin B12 enriched mushroom powder, vitamin D enriched mushroom powder and
-
der exhibited higher NOSA compared to commercially available mushroom powders.
Figure 3: Nitric oxide scavenging activity of mushroom powders.
ABTS radical scavenging activity (ABTS)
ABTS radical scavenging activity was measured as trolox equivalent and inhibitory concentration of the powder (IC50) required for
50 indicates better antioxidant activity.
Citation: Supriya Yadav and Jude Wilson. “Antioxidant and Ergothioneine Assessment of Nutritionally Enriched Agaricus bisporus
Powders”. EC Nutrition 17.2 (2022).
Antioxidant and Ergothioneine Assessment of Nutritionally Enriched Agaricus bisporus Powders
Whole mushroom powder50 compared to Batch 1 (2.81 mg) and Batch 3
50. These results indicated consistent retention of antioxi-
Vitamin D enriched mushroom powder50 compared to Batch 2 (2.99 mg)
50 compared to Batch 2. Batch 1 and Batch 3 exhibited
50. Overall, Batch 1 exhibited better antioxidant activity compared to other batches of vitamin D en-
riched mushroom powder (Figure 4 and table 3).
Vitamin B12 enriched mushroom powder50 compared to Batch 2. Batch 2
5050
strong ABTS scavenging activity indicating strong antioxidant activity.
Selenium enriched mushroom powder50
0.02) IC50 compared to Batch 3. Although, variability across the three batches was observed for ABTS radical scavenging activity, Batch
3 of the selenium enriched mushroom powder exhibited the best ABTS scavenging activity among all mushroom powders (Figure 4 and
table 3).
the lowest IC50 indicating the highest ABTS radical scavenging activity followed by the vitamin B12 enriched mushroom powder. However,
(Figure 4).
Comparison of nutritionally enriched mushroom powders vs marketed available mushroom powders: Finally, nutritionally en-
riched Agaricus bisporus50 compared to commercial powders (CP1 (6.82
better antioxidant activity compared to the commercial mushroom powders (Figure 4).
Figure 4: ABTS radical scavenging activity of mushroom powders (Lower the IC50 value better is the antioxidant activity).
Citation: Supriya Yadav and Jude Wilson. “Antioxidant and Ergothioneine Assessment of Nutritionally Enriched Agaricus bisporus
Powders”. EC Nutrition 17.2 (2022).
Antioxidant and Ergothioneine Assessment of Nutritionally Enriched Agaricus bisporus Powders
Hydroxyl radical scavenging activity (OHSA)
Whole mushroom powder-
of strong antioxidant activity was observed among WMP batches (Figure 5).
Vitamin D enriched mushroom powder
vitamin D enriched mushroom powder.
Vitamin B12 enriched mushroom powder
2 (Figure 5). This indicated a strong retention of OHS activity in MBio’s vitamin B12 enriched mushroom powder.
Selenium enriched mushroom powder
the selenium enriched mushroom powders exhibited strong retention of OHS activity.
Overall, selenium enriched mushroom powder (25.52 mg/g) and the vitamin B12 enriched mushroom powder (25.68 mg/g) followed
(18.28 mg/g).
Comparison of nutritionally enriched mushroom powders vs marketed available mushroom powders: The selenium enriched mush-
Chaga (CP2 12.34 mg/g), Reishi (CP3 17.09 mg/g), Lion’s Mane (CP4 12.18 mg/g) and a mixture of mushroom powders (CP5 11.06
mg/g)) (Figure 5). Finally, Agaricus bisporus
radical scavenging activity compared to marketed mushroom powders.
Figure 5: Hydroxyl radical scavenging activity of mushroom powders.
Citation: Supriya Yadav and Jude Wilson. “Antioxidant and Ergothioneine Assessment of Nutritionally Enriched Agaricus bisporus
Powders”. EC Nutrition 17.2 (2022).
Antioxidant and Ergothioneine Assessment of Nutritionally Enriched Agaricus bisporus Powders
Ergothioneine analysis
Ergothioneine analysis was conducted at CampdenBRI using LC/MS/MS. The vitamin D enriched mushroom powders (average 1303
content compared to the vitamin B12 enriched mushroom powder and whole mushroom powders (WMP). Ergothioneine levels for WMP
ranged from 547 - 1009 mg/Kg (average 855 mg/Kg) and the vitamin B12 enriched mushroom powders ranged from 189.5 - 827 mg/Kg
(average 519.3 mg/Kg) (Figure 6 and table 3). The marketed mushroom powders exhibited ergothioneine content as; shitake (CP1 95.5
mg/Kg), Chaga (CP2 1408 mg/Kg), Reishi (CP3 166 mg/Kg), Lion’s Mane (CP4 20 mg/Kg) and a mixture of different mushroom powders
(CP5 35 mg/Kg) (Figure 6).
Figure 6: Ergothioneine content of mushroom powders.
Discussion
Agaricus bisporus mushroom derived powder products and randomly selected
marketed mushroom powders were compared for their in vitro antioxidant activity. The antioxidant activity was represented as equiva-
lent to standard antioxidants, such as Trolox and gallic acid.
by the vitamin B12 enriched mushroom powder, compared to vitamin D enriched mushroom powder and whole mushroom powder.
suggest that the selenium enriched mushroom powder may contain higher antioxidant polyphenolic molecules responsible for free radi-
known antioxidant molecule. Therefore, these results suggest that selenium enrichment of the Agaricus bisporus mushrooms might have
resulted into higher antioxidant content of the product. This result is in agreement with the literature that reported increased antioxidant
activity in mushrooms as a result of increased selenium uptake by mushrooms, which are cultivated on selenium hyper-accumulated agri-
Citation: Supriya Yadav and Jude Wilson. “Antioxidant and Ergothioneine Assessment of Nutritionally Enriched Agaricus bisporus
Powders”. EC Nutrition 17.2 (2022).
Antioxidant and Ergothioneine Assessment of Nutritionally Enriched Agaricus bisporus Powders
content suggesting that the vitamin D and ergothioneine may play a role as an antioxidant in the vitamin D enriched mushroom powders.
Agaricus bisporus mushroom powder to higher levels (8000 IU/g)
-
ders derived from freshly harvested whole mushrooms processed under mild conditions to preserve its nutrient rich content. Mushrooms
contain proteins, polysaccharides, terpenes, terpenoids, and important antioxidants including polyphenols (phenolic acids) glutathione
-
Therefore, MBio’s mushroom powders remain a source for important nutrients and bioactive ingredients, such as polyphenols and ergo-
thioneine, which are known antioxidants that might explain the higher antioxidant activity of MBio’s all mushroom powders. However,
other marketed mushroom powders contains different mushroom strains including Shitake, Chaga, Reishi, Lion’s Mane and a mixture of
different mycelial cultures of various strains of the mushrooms. This might explain the different antioxidant activity observed in markedly
available mushroom powders compared to MBio’s Agaricus bisporus derived and nutritionally enriched mushroom powders. Literature
-
ever, antioxidant activities of fresh or dried A. bisporus have been reported. Total polyphenols from white A. bisporus mushrooms after
-
ders ranged from 5 - 6 mg/g dry weight, which are in agreement with the above literature.
compared to other nutritionally enriched (vitamin D and vitamin B12) mushroom powders and marketed mushroom powders such as
shitake, reishi, lion’s mane and a mixture of different mushroom powders (CP1, CP3, CP4, and CP5), respectively. Only commercial Chaga
powder (CP2 1408 mg/Kg) exhibited higher ergothioneine content compared to other commercial mushroom powders and nutrition-
ally enriched mushroom powders such as: vitamin D enriched mushroom powder (1303 mg/Kg), selenium enriched mushroom powder
-
between polyphenols and ergothioneine. These results are in agreement with Dubost., et al
antioxidant activity might be independent of other radical scavenging antioxidant activities. However, this does not affect the biological
activity of ergothioneine and overall antioxidant activity of MBio’s nutritionally enriched mushroom powders.
Ey., et alet alA. bisporus (white) and A. bisporus
(brown) as 4.6 and 9.3 mg/Kg dry weight, respectively. Additionally, Shiitake mushroom fruiting body indicated higher ergothioneine con-
tent of 2840 mg/Kg dry weight. Some of the mushroom mycelial samples including Ganoderma lucidum (Reishi), Grifola frondosa (Mai-
take) and Hericium erinaceus
Similarly, Inonotus obliquus (Chaaga) mushroom mycelia also reported higher ergothioneine content of 252.1 mg/Kg dry weight. How-
Citation: Supriya Yadav and Jude Wilson. “Antioxidant and Ergothioneine Assessment of Nutritionally Enriched Agaricus bisporus
Powders”. EC Nutrition 17.2 (2022).
Antioxidant and Ergothioneine Assessment of Nutritionally Enriched Agaricus bisporus Powders
ever, Dubost., et alA. bisporus white button and portabellas as 210 mg/Kg and 450 mg/
Kg, respectively. Furthermore, the authors reported the ergothioneine content in specialty mushrooms, including shiitake (1980 mg/Kg),
-
room types, which might be related to different type of raw materials used (fruiting body or mycelia), difference in production, drying
methods, and various extraction and analysis methods used. However, MBio’s nutritionally enriched and whole mushroom powders are
developed using A. bisporus
suggest that MBio’s production process is optimal for the retention of higher levels of ergothioneine in all MBio’s mushroom powders.
The current study reported variable levels of ergothioneine across batches and types of mushroom powders. However, this variability
can be explained on the basis of sourcing of raw material for each batch and homogeneity of the ergothioneine in the mushroom pow-
ders. This is also supported by the literature. Dubost., et al
pathway is also affected by various stress factors exposure to mushroom mycelia such as dry compost to the crops, which resulted in
compared and reported by Zhang., et alet al
produced by mushrooms.
Agaricus bisporus derived nutri-
tionally enriched mushroom powders are a good source of ergothioneine (which is a known antioxidant) compared to commercially
available Shitake, Reishi, Lion’s Mane and other mixtures of different mushroom powders.
Conclusion
Conclusively, all nutritionally enriched Agaricus bisporus -
available mushroom powders. Furthermore, whole mushroom powder developed using Agaricus bisporus
that MBio’s nutritionally enriched Agaricus bisporus
to prevent the progression of various diseases, caused by free radicals, such as certain cancers and might play a role in improvement of
immune health.
Acknowledgements
Group.
SY: Conceived and designed the experiments, performed the experiments, analysed and interpreted data and wrote the manuscript,
JW: guided the work, interrogated results, and interpretation of the data and reviewed the manuscript. The authors of this article declare
Citation: Supriya Yadav and Jude Wilson. “Antioxidant and Ergothioneine Assessment of Nutritionally Enriched Agaricus bisporus
Powders”. EC Nutrition 17.2 (2022).
Antioxidant and Ergothioneine Assessment of Nutritionally Enriched Agaricus bisporus Powders
1. Mayne ST. “Oxidative Stress, Dietary Antioxidant Supplements, and Health: Is the Glass Half Full or Half Empty?” © 2013 American
Association for Cancer Research”. Cancer Epidemiol Biomarkers and Prevention 22.12 (2013): 2145-2147.
2. Halliwell B. “Free Radicals and Other Reactive Species in Disease”. ELS (2015).
3. Kadiiska M., et al. “Biomarkers of Oxidative Stress Study Ii: Are Oxidation Products Of Lipids, Proteins, And Dna Markers Of Ccl 4
Poisoning?” Free Radical Biology and Medicine 38.6 (2005): 698-710.
4. Uttara B., et al. “Oxidative Stress and Neurodegenerative Diseases: A Review of Upstream and Downstream Antioxidant Therapeutic
Options”. Current Neuropharmacology 7.1 (2009): 65-74.
5. et alJour-
nal of Central European Agriculture 11.4 (2010): 387-392.
6. Nagmoti D., et al. “Antioxidant Activity and Free Radical-Scavenging Potential of Pithecellobium Dulce Benth Seed Extracts”. Free
Radicals 2.2 (2011): 37-43.
7. Free Radical Research Communications 18.4 (1993): 195-199.
8. Rathee S., et al. “Mushrooms as Therapeutic Agents”. Brazilian Journal of Pharmacognosy 22.2 (2012): 459-474.
9. Feeney M., et al. “Mushrooms and health summit proceedings”. Journal of Nutrition 144.7 (2014): 1128S-1136S.
10. Ayeka PA. “Potential Of Mushroom Compounds As Immunomodulators In Cancer Immunotherapy: A Review”. Evidence-Based Com-
plementary and Alternative Medicine (2018): 9.
11. Wang Q., et al. “Bioactive Mushroom Polysaccharides: A Review on Monosaccharide Composition, Biosynthesis And Regulation”.
Molecules 22.6 (2017): 955.
12. Xu X., et al. “Bioactive Proteins From Mushrooms”. Biotechnology Advances 29.6 (2011): 667-674.
13. Kalaras MD., et al. “Mushrooms: A rich source of the antioxidants ergothioneine and glutathione”. Food Chemistry 233 (2017): 429-
433.
14. Sánchez C. “Bioactives From Mushroom And Their Application. Chapter 2, ©Springer International Publishing AG 2017M. Puri (edi-
tion.)”. Food Bioactives (2017): 23-57.
15. Hartman PE., et al. “Ergothioneine, Histidine, and Two Naturally Occurring Histidine Dipeptides as Radioprotectors against
Radiation Research 114.2 (1988): 319-330.
16. Paul B and Snyder S. “The unusual amino acid L-ergothioneine is a physiologic cytoprotectant”. Cell Death Differ 17.7 (2010): 1134-
1140.
17. Dubost N., et al -
troscopy”. International Journal of Medicianl Mushrooms 8.3 (2006): 215-222.
18. Dubost N., et al-
pacity”. Food Chemistry 105.2 (2007): 727-735.
Bibliography
Citation: Supriya Yadav and Jude Wilson. “Antioxidant and Ergothioneine Assessment of Nutritionally Enriched Agaricus bisporus
Powders”. EC Nutrition 17.2 (2022).
Antioxidant and Ergothioneine Assessment of Nutritionally Enriched Agaricus bisporus Powders
19. Dubost N., et al -
room Agaricus bisporus (J. Lge) Imbach (Agaricomycetideae)”. International Journal of Medicianl Mushrooms 9.2 (2007): 163-176.
20. Rahman MM., et al-
gladesh”. BMC Research Notes 8 (2015): 621.
21. Gite S., et al. “Antioxidant quality ranking of some plant materials based on multiple in vitro radical scavenging assays”. Journal of
Chemical and Pharmaceutical Research 7.7 (2015): 152-159.
22. Alger M. “Polymer Science Dictionary”. Springer (1997): 152.
23. Ariga T and Hamano M. “Radical Scavenging Action And Its Mode in Procyanidins B-1 And B-3 From Adzuki Beans To Peroxyl Radi-
cals”. Agricultural and Biological Chemistry 54.10 (1990): 2499-2504.
24. Chandra S., et al. “Assessment Of Total Phenolic And Flavonoid Content, Antioxidant Properties, And Yield Of Aeroponically And Con-
Evidence-Based Complementary and Alternative Medicine
(2014): 253875.
25. et al. “Studies On Antioxidant Activities Of Desmodium Gangeticum”. Biological and Pharmaceutical Bulletin 26.10
(2003): 1424-1427.
26. et al. “Antioxidant Capacity Of Thermally – Treated Buckwheat”. Polish Journal of Food and Nutrition Sciences 57.4 (2007):
465-470.
27. Reis FS., et al
study between in vivo and in vitro samples”. Food and Chemical Toxicology 50.5 (2012): 1201-1207.
28. Prior RL., et al. “Standardized methods for the determination of antioxidant capacity and phenolics in foods and dietary polyphenols
Journal of Agricultural and Food Chemistry 53 (2005): 4290-
4302.
29. Bhatia P., et al. “Enhanced Anti-oxidant Activity as a Function of Selenium hyperaccumulation in Agaricus bisporus Cultivated on Se-
rich Agri-residues”. Journal of Biologically Active Products from Nature 4. 5-6 (2014): 354-364.
30. Bulletin of the National Research Centre 44.34
(2020).
31.
cholesterol, ergosterol and tamoxifen and relevance to anticancer action”. FEBS Letters 326.1-3 (1993): 285-288.
32. Sharpe E., et al. “Comparison of antioxidant activity and extraction techniques for commercially and laboratory prepared extracts
from six mushroom species”. Journal of Agriculture and Food Research 4 (2021): 100-130.
33. et al
species of Agaricus”. European Food Research and Technology 244 (2018): 259-268.
34. Palacios I., et al. “Antioxidant properties of phenolic compounds occurring in edible mushrooms”. Food Chemistry 128 (2011): 674-
678.
Citation: Supriya Yadav and Jude Wilson. “Antioxidant and Ergothioneine Assessment of Nutritionally Enriched Agaricus bisporus
Powders”. EC Nutrition 17.2 (2022).
Antioxidant and Ergothioneine Assessment of Nutritionally Enriched Agaricus bisporus Powders
35. Gan C., et al. “Antioxidant analysis of different types of edible mushrooms (Agaricus bisporous and Agaricus brasiliensis)”. Interna-
tional Food Research Journal 20.3 (2013): 1095-1102.
36. Reis F., et al. “Analytical methods applied to the chemical characterization and antioxidant properties of three wild edible mushroom
species from northeastern Portugal”. Food Analytical Methods 7 (2014): 645-652.
37. Liu J., et al. “In vitro and in vivo antioxidant activity of ethanolic extract of white button mushroom (Agaricus bisporus)”. Food and
Chemical Toxicology 51 (2013): 310-316.
38. Ey J., et al. “Dietary sources and antioxidant effects of ergothioneine”. Journal of Agricultural and Food Chemistry 55.16 (2007): 6466-
6474.
39. Bao H., et al Journal of the Science of
Food and Agriculture 90.10 (2010): 1634-1641.
40. Chen S., et alLWT - Food
Science and Technology 47.2 (2012): 274-278.
41. Zhang W., et al. “Aqueous Extraction of Ergothioneine from Mycelia of Pleurotus ostreatus and Ergothioneine Accumulation Regular-
ity during Submerged Fermentation”. Research and Reviews: Journal of Microbiology and Biotechnology 5.1 (2016):2347-2286.
Volume 17 Issue 2 February 2022
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