Effect of modified atmosphere packaging on selected functional characteristics of Agaricus bisporus


Mushrooms are proven as a functional food due to their numerous beneficial effects on human health. Contemporary consumers purchase cultivated mushrooms that spend some time on the shelf, which makes it questionable whether this food can still be called “functional”. An examination was performed on selected characteristics of white and brown (Portobello) Agaricus bisporus stored in commercial (air) and three modified atmosphere packagings (MAP): high nitrogen, low carbon dioxide and low oxygen packaging. The amount of ascorbic acid decreased throughout 22 days for both varieties, especially in the white variety stored in commercial packaging. Similarly, total flavonoids decreased, although not to a significant degree. Metal chelating ability was pronounced throughout the storage period, with minor changes in the case of the brown variety. Diabetes-connected enzymes were inhibited by A. bisporus, while inhibition was significantly higher toward α-amylase. Nitrogen-rich packaging suppressed α-amylase and stimulated α-glucosidase in the white strain. Both the commercial packaging and the MAP samples exhibited changes in their functional characteristics over three weeks of cold storage. MAP, especially the low oxygen packaging, provided the best option for the preservation of the majority of functional characteristics examined in this research. Enzymes activities appeared to be more specifically tuned, and dependent on parameters not covered here. Brown variety was more resistant to environmental changes with respect to its functional characteristics.

Graphic abstract


Agaricus bisporus is one of the most widely cultivated mushroom species and comes to the fourth place based on the worldwide produced volume per year (15%) [1, 2]. However, the white button mushroom undergoes fast deterioration after its harvest and after one to three days at room temperature, it is no longer suitable for the market [3]. Globally, 45% of all produced mushrooms go to the market as fresh, where in Europe it is almost doubled (80%), so it is not surprising that advanced technologies which diminish economical losses and prolong the mushroom's shelf life and quality are needed [4, 5]. In a review by Zhang et al. [3] there is an extensive look into the traditional methods with some novel modifications as well as into the contemporary industrial-scale solutions like modified atmosphere packaging, MAP, ultrasound or the electrolyzed water that can be applied to prolong the shelf life of mushrooms. Still, MAP is considered as the most efficient, easy to perform, and an economical method for fresh mushroom preservation.

The United States International Trade Commission has announced a rise of the mushroom market, especially the mushrooms stored in innovative packages [6]. Another fact that has a strong influence on this market is the increase in the number of so-called “health consumers” who apprize the functional/medicinal benefits of food [5]. Although white button mushroom is considered as a high-quality food that even positively affects cognitive function in humans and express anti-obesity activity [7], the inevitable deterioration of the fruit body during the shelf life must affect its functionality as well.

Most of the current studies are focused on sensory and nutritional changes that occur in mushrooms during the post-harvest period [8,9,10]. On the other side, A. bisporus possesses bioactive compounds with therapeutic properties [11], so it can be considered as a functional food. The question arises as to what happens to the functional characteristics of A. bisporus if stored for a long time? Moreover, how does the button mushroom functionality aspect change when stored in MAP?

Aiming to clarify the issues important for mushroom purchasing, especially as a part of the functional food market, this study explores several functional characteristics of white and brown (Portobello) A. bisporus when stored in three types of MAP during 22 days at the temperature of 4 °C. Namely, we tested the levels of ascorbic acid and flavonoids given the fact that they are common antioxidants in mushrooms [12]. Based on the connection, and especially the correlation between antioxidant compounds and metal ion chelating ability, we also tested the Fe2+ chelation activity. Other methods for antioxidant activity evaluation such as 2, 2-azinobis-(3-ethylbenzothiazoline-6-sulfonate radical scavenging activity was evaluated in our previous study [13]. Recently, it was confirmed in animal studies that button mushroom might be beneficial in treating diabetes [14, 15]. Therefore, we examined in vitro changes in samples activity toward two diabetes-connected enzymes, α-amylase and α-glucosidase. Selected characteristics were followed for 22 days based on our previously published studies which tested a wide range of changes during shelf life and proved that prolonged MAP can positively affect the nutritional value as well as microbiological profile (safety) [13]. Similar was observed by Gholami et al. [8]. No pathogenic bacteria such as Clostridium botulinum has been found in our samples. To the best of our knowledge, this is the first study that screens changes of the selected functional characteristics of two types of A. bisporus stored in MAP.

Material and methods


Methanol, 2,6-dichlorphenolindophenol (DFIF), metaphosphoric acid, catechin, NaNO2, AlCl3, NaOH, FeSO4, ferrozine, EDTA, phosphate buffer (pH 7.5), DMSO, 4-nitrophenyl-α-D-glucopyranoside (PNPG), α-glucosidase (EC, from Saccharomyces cerevisiae), phosphate buffer (pH 6.0), α-amylase (EC, from Bacillus subtilis), 2-chloro-4-nitrophenyl-α-D-maltotrioside (CNP-G3) were purchased from Sigma Aldrich (St. Louis, MO, USA).

Mushrooms and packaging

In this study, freshly harvested fruit bodies of white button mushroom and the brown strain (Portobello) (A. bisporus, Italspawn, strains F599 and FB30, commercial varieties) were purchased from the same local producer in Belgrade, Serbia. All samples were cultivated using chicken litter-based compost (standardized), originated from the same growing tunnel, the same cycle, and were harvested during the first flush. A cap diameter was 30–70 mm. After being harvested, the mushrooms were transferred to a laboratory in a refrigerated vehicle and stored at 4 °C before packaging.

Only the carpophores free from defects (scars, blemishes, not whole, not healthy or good looking), were selected for the experiment. The characteristics of packaging material were: 85 mm thick (PA/PE/PE) bags, transmission rates of 60 mL O2, 12 mL N2, 180 mL CO2 m−2 d−1 atm−1, size of 200–300 mm. The bags were packed and sealed with a HVC-510 T/2A packaging machine. All bags were the same weight (250 g).

Active modified packaging conditions were: N2 (Gourmet N, 100% N2), CO2 (Gourmet N70, 30% CO2 and 70% N2) and O2 (30% O2 and 70% N2). All gas-combinationes were supplied by Messer Tehnogas (Belgrade, Serbia) as already prepared (food industry grade). Every first packaging was checked with gas analyzer (Oxybaby, Witt-Gastechnik GmbH & Co KG, Witten, Germany), while the last packaging was checked and verified to confirm that all conditions were as defined. Control sample contained only air to simulate the common commercial packaging. After being sealed, all packages were stored at 4 °C and relative humidity 95% for 22 days. Analyses were done on the 5th, 8th, 12th, 15th, 19th and 22nd day.

Mushroom extract

Mushroom samples were cut into thin slices and lyophilized (Christ BETA 2–8 LD plus freeze dryer, Osterode, Germany), powdered and subjected to methanol extraction. The solvent of choice (methanol) favors the extraction of compounds analyzed in this study and it is readily available. Sample (10 mg) was mixed with methanol (200 mL) and macerated for 24 h, on a magnetic stirrer (1 × g) at room temperature. Then, the material was filtered through Whatman No. 4 and the liquid portions were collected. The procedure was repeated three times and liquid parts were combined before low pressure evaporation (Büchi Rotavapor RII, Flawil, Switzerland) at 37 °C. The dry extracts were kept in the refrigerator at 4 °C before analysis. All results are expressed based on dry extract mass.

Ascorbic acid content

The method described by Barros et al. [16] was adjusted for a 96-well microplate assay. The extract (50 mg) was dissolved in 1% metaphosphoric acid (1 mL) and macerated for 20 min on a magnetic stirrer at room temperature. The sample was then centrifuged (six min, 18,000 × g) and an aliquot of the supernatant was mixed with 240 µL of DFIF. The absorbance was read after six min at 490 nm (Microplate reader Biohit ELx808, BioTek, Winooski, US; the same reader was used in all other methods) against a blank (sample with 1% metaphosphoric acid). All samples were analyzed in triplicates and calculated based on the standard curve of L-ascorbic acid (concentration range: 0.03125–0.25 g L−1). The results are expressed as g kg−1 of ascorbic acid (based on dry extract mass).

Total flavonoid content

Quantification of total flavonoids in mushroom samples was done based on colorimetric microplate assay [17]. The mushroom extract (5 g) was dissolved in methanol and added to the microplate well in the amount of 25 µL. After the addition of 125 µL of redistilled water and 7.5 µL of 5% NaNO2 the mixture was incubated for five min at room temperature. Then, 15 µL of 10% AlCl3 were added and the plate was incubated for another 5 min at room temperature. Finally, 50 µL of 1 M NaOH and 27.5 µL of redistilled water were added and the absorbance (510 nm) was measured immediately. Prior to the absorbance reading the microplate was mixed for 10 s. Similarly, a blank was prepared by adding methanol instead of the mushroom extract solution. Standard catechin solutions (0.0625–2 g L−1) were used to construct the standard curve. The results are expressed as mean ± standard deviation (g kg−1 catechin equivalents). All measurements were performed in triplicates.

Fe2+ chelation ability

The ability of mushroom samples to chelate Fe2+ ion was tested based on microplate assay by Santos et al. [18]. 160 µL of redistilled water together with 20 µL of 0.3 mM FeSO4 were added to the mushroom extract (5 mg) dissolved in methanol, and incubated for five min. Then, 30 µL of 0.8 mM ferozine solution were added. The plate was then incubated for 15 min at room temperature and the absorbance was measured at 490 nm against a blank (sample contained all components except the mushroom extract, which was replaced with methanol). The results are expressed as % of Fe2+ chelation. The measurements were performed in triplicates.

α-amylase inhibitory activity

In-vitro α-amylase inhibition assay adjusted for microplates was conducted according to the previously described method [19]. The mushroom extract was dissolved in 5% DMSO and 10 µL of its solution were added into the plate as well as 90 µL of phosphate buffer (0.1 M, pH 6.0, containing 0.02% NaN3). The next step was the addition of 80 µL of enzyme solution prepared in phosphate buffer (the concentration of α-amylase in the microplate well was 0.05 U mL−1). At the end 20 µL of CNP-G3 were added. The reaction was monitored throughout 30 min at a wavelength of 405 nm; the value was recorded every three min. The ability of mushroom samples to inhibit α-amylase was calculated based on equation:

$$ \% {\text{ inhibition }} = \, \left( {{\text{Slope}}_{{{\text{blank}}}} - {\text{ Slope}}_{{{\text{sample}}}} } \right)/{\text{Slope}}_{{{\text{blank}}}} \times \, 100. $$

In-vitro α-glucosidase inhibitory activity

A microplate assay described by Kirakosyan et al. [19] was used to determine the A. bisporus ability to inhibit α-glucosidase. 90 µL of 0.1 M phosphate buffer (pH 7.5) and 10 µL of the mushroom extract sample dissolved in methanol were added into the microplate. The enzyme was then added (80 µL) so that its concentration in the well was 0.08 U mL−1 followed by the addition of PNPG (20 µL). The colorimetric reaction was followed for 35 min with reading on every 30 sek at 405 nm. The results were calculated according to the same equation as above:

$$ \% {\text{ inhibition }} = \, \left( {{\text{Slope}}_{{{\text{blank}}}} - {\text{ Slope}}_{{{\text{sample}}}} } \right)/{\text{Slope}}_{{{\text{blank}}}} \times \, 100 $$

Statistical analysis

All measurements were performed in triplicates and the results were given as mean values ± standard deviation. Statistica software 12.0 (Statistica, Tulsa, OK, SAD) was used for data processing: one-way analysis of variance and Tukey’s HSD post hoc test (p < 0.05).

Results and discussion

Ascorbic acid content

The level of ascorbic acid in both A. bisporus strains is presented in Table 1. The white variety contained a significantly higher level of ascorbic acid (1.28–5.51 g kg−1 ascorbic acid) followed by brown (0.54–1.88 g kg−1 ascorbic acid) throughout the whole storage period. Barros et al. [12] reported much lower contents (0.02–0.04 g kg−1 of extract), while others obtained higher results but for fresh or dry samples, and not extracts. The difference in the amount of ascorbic acid has already been noticed in mushrooms and is ascribed to factors such as geographical origin, maturity stage, genotype and climate conditions [20].

Table 1 Levels of ascorbic acid and flavonoids in white and brown A. bisporus packed in air, O2, CO2 and N2-rich MAP measured over a period of 22 d of cold storage (4 °C)

Generally, mushrooms are not considered as rich sources of ascorbic acid. However, fungi produce erythroascorbate, an analog of ascorbate through of at least three biosynthetic pathways which point to its physiological role for the mushroom itself [20,21,22]. According to Georgiou et al. ascorbate caused a reduction of lipid peroxidation in a sclerotial strain of fungi, being a mechanism of oxidative stress defense [23]. And although present in amounts that are not especially high, vitamin C act as a synergist for other antioxidants thus adding to a mushroom’s contribution to a functional diet for humans [20].

The modified atmosphere induced a significant lowering in ascorbic acid level in the first week of the storage in both strains, a fact also reported by Li and Zhang [24]. At the same time, white A. bisporus packed in commercial package (filled with air) resisted this trend, but only for a short period. An elevated concentration of O2, N2, and CO2 generated a stress-environment that disturbed normal metabolism and forced the fruit bodies to activate its reactive oxidative species-defense mechanism. The O2 and N2 MAP were particularly stressful for the first eight days of cold storage. Interestingly, in the second week of the storage, a very pronounced increase in ascorbic acid concentration was observed in all packagings with the white strain, and especially in the mushrooms packaged inr MAP with CO2 and O2. This increase was present but not so prominent in Portobello. However, further storage resulted in additional further reduction, so that the ascorbic acid level was lower than that at the beginning of the storage. The observed trend is in agreement with the findings from other studies dealing with fruit and juice stored in MAP [25, 26]. This behavior was expected since ascorbic acid is the most sensitive compound in fresh mushrooms, especially vulnerable under prolonged storage and changed storage conditions, as in our experiment [24]. After three weeks of cold storage the level of ascorbic acid in both strains was the highest in O2 MAP. The difference between O2 and other MAPs was statistically significant. This could be due to the decreased respiratory metabolism as proposed by Liu et al. [27].

By comparing the changes in MAP, significant differences were observed for the two A. bisporus varieties, pointing that although they belong to the same species these two strains have different metabolism behaviors.

Total flavonoid content

As is evident from the Table 1 the amount of flavonoids decreased during 22 days of storage, similarly for both mushroom varieties. The average amount of flavonoids was about 1 g kg−1 catechin equivalents which corresponds to the findings of Palacios et al. [28] and Kaewnarin et al. [29]. Significant changes occurred in the commercial packaging of the white button mushroomafter five days of storage, which was not the case with the brown A. bisporus. The most drastic change appeared in O2 MAP (white variety) and CO2 package (brown variety); it stimulated flavonoid accumulation in both varieties but at different storage times (during the first week in white and the second in Portobello). Gąsecka et al. [30] pointed that excess of glucose serves as a reservoir for enhanced synthesis of secondary metabolites, such as phenolic compounds. When connected with our previous work [13], total sugar and flavonoid content were not correlated. However, this should be taken with caution since the method we applied for measuring sugar content has its limitations, like non-selectiveness for different types of carbohydrates present in fungi. The phenol–sulfuric method that we used cannot detect the presence of all sugar forms. This might affect the inconsistencies in the consequent sugar-phenolics correlation. Instead, chromatographic methods might be used to enable a wider range of detection. Methods like high-performance liquid chromatography might be a better choice since they are rapid, simple, and specific [31]. Unfortunately, before the analysis the samples have to be prepared: nonspecifically hydrolyzed with acids (HCl, H2SO4, CrH4O8S2). Another popular choice is enzymatic assays characterized by high specificity and sensitiveness. However, no one enzyme kit can cover all sugars occurring in mushrooms and there is the need to couple it with spectrophotometry [32]. Recently high-performance liquid chromatography coupled to corona charged aerosol detection was proposed as a method with high sensitivity but only for the determination of free sugars and mannitol in mushrooms [33]. The question is an also-which type of phenolic compounds are generated and under which conditions from present glucose. Secondary metabolites, like phenolic compounds, are formed under unfavorable conditions as an organism’s stress response. Different types of stress will stimulate different secondary metabolites like Yan et al. reported for Pleurotus ostreatus exposed to heat stress [34]. The authors identified accelerated glycolysis and the tricarboxylic acid cycle. Similarly, Sudheer et al. measured the increase of total phenolics, flavonoids, polysaccharides, and ganoderic acids when medicinal mushroom Ganoderma lucidum was exposed to external CO2 [35]. Additionally, flavonoids undergo oxidation when exposed to stress so the initial molecule becomes a mixture of compounds with different structural features and might become invisible for a simple screening assay [36]. Another reason for flavonoid changes in our samples might be the fact that they build complexes with proteins [37]. And proteins do go through the degradation process induced by prolonged storage and MAP influence, as we previously proved [13]. Furthermore, the presence of flavonoid-protein compounds in mushrooms restricts the extraction of flavonoids, so that the total amount might be underestimated over time.

Although the white button and Portobello mushroom belong to the same species, genetic differences affected the stress reaction and flavonoid levels in these two strains.

Flavonoid content was similar for both mushroom strains and the highest level was measured after eight days of cold storage in MAP with O2.

Antioxidant activity

Flavonoids are widely present in nature, especially in plants, but mushrooms also contain this group of phenolic compounds [29, 38]. They are considered as antioxidants, especially active in metal chelating, among other mechanisms of protection against reactive oxygen species [11]. Samples tested in our work showed high metal chelating potential at a concentration of 5 g L−1, during the entire period of 22 days of storage, and in all MAPs (Fig. 1). Chelating ability was similar for both strains over prolonged shelf life. All MAPs except O2 caused a decrease in the white strain chelating ability in a statistically significant degree. CO2 packaging influenced the most pronounced decrease, while O2 appeared as the most stable, variating less than 10%. Interestingly, the white variety experienced a sudden drop on the 19th day for all MAP examined in this study, and the commercial atmosphere packaging showed the highest percentage of the loss of antioxidative ability. On the other side, Portobello tended to be a more stable chelating agent for 22 days of cold storage (between 40 and 60%). Its metal chelating ability ranged from 40 to 65% with a wave-like pattern. Piljac-Žegarac et al. [39] reported similar behavior in case of pomegranate, strawberry, blueberry, and cranberry juice whose antioxidant activity was assayed by 2,2-diphenyl-1-picrylhydrazyl radical scavenging method. N2 MAP went through antioxidant ability decrease in the first week of cold storage, which was not the case with other MAPs. However, this MAP induced the most stable metal ion chelation over a tested period. O2 acted stimulating for almost two weeks with a significant and sudden drop of antioxidant activity from day 12 to the end of the experiment. In the brown variety standard packaging (air) showed to be the best not just in preserving antioxidant ability but even to increase it.

Fig. 1

Fe2+ chelation ability of the white and the brown A. bisporus stored in MAP. Ferrous ion chelating ability of a the white A. bisporus in MAP filled with air, O2, CO2 and N2; b the brown A. bisporus in MAP filled with air, O2, CO2 and N2 during 22 days of cold storage (4 °C). All results are means of three replicates ± standard deviation

When connected with total flavonoid content it has been noticed that the drop in flavonoid content was followed by the drop in the chelation ability.

However, the flavonoid decrease was not as pronounced, implying that there are other compounds that also affect this type of antioxidant activity. Another, probably more significant cause might be the change in the flavonoid profile. Some flavonoids like kaempferol or apigenin, which were identified in A. bisporus, are very stable when exposed to alkaline decomposition or enzymatic treatment. Others, such as myricetin, decomposed very fast (4–25 min) [11, 29]. It has been proved that the products of phenolic compounds decomposition (as well as those of flavonoids) save its antioxidant ability in significant percentage, which was probably the case in metal chelation as well. Total phenolic compounds in A. bisporus did not fit into this pattern: their amount did not correlate with the metal chelation ability, which was the proof of the importance of flavonoids when it comes to the overall antioxidant activity of mushrooms [13, 40].

Inhibition of type 2 diabetes mellitus related enzymes

Mushrooms, including A. bisporus, are already recognized as functional food as well as foods that have beneficial effect in preventing and controlling diabetes [41]. There are two enzymes linked with diabetes regulation, α-amylase and α-glucosidase [42], which inhibition we examined (Fig. 2). All tested samples exhibited a significant inhibition of α-amylase activity (maximal measured % of inhibition was 92.97%). The white strain had a lower ability to inhibit α-amylase when compared with the brown one, and the trend was more extreme (Fig. 2a). During the first week of the storage, the % of α-amylase activity increased in all samples except for N2, and then significantly decreased. The middle of the storage period was the time at which all samples (except N2) showed the highest inhibition of α-amylase (86–88%). On the other side, N2 strongly suppressed the ability of A. bisporus to inhibit this enzyme. The highest fluctuations of enzyme inhibiting activity were observed in O2 packaging. The low oxygen package had the highest values for total phenolic compounds which were reported as being responsible for α-amylase inhibition in methanol extracts of plants [40, 43]. However, no clear trend can be observed: in most of the cases, the increase in total phenolic compound content was followed by the increase in α-amylase inhibition. Inconsistencies present here might be explained through the qualitative profile of phenolics present in mushrooms. The sum of phenolic compounds does not have to be equally potent as some of the specific molecules present in the sample. Furthermore, other biologically active compounds, not covered by this research, might be responsible for the observed trends. Additionally, the decrease in O2 concentration affects cell respiration: O2 is a substrate in cell respiration and during this process, intermediary molecules necessary for secondary metabolites (like phenolic compounds) formation are constituted [44]. So, less of these compounds are present under decreased O2 concentration.

Fig. 2

α-amylase and α-glucosidase inhibitory activity of the white and the brown A. bisporus stored in MAP. Inhibition of α-amylase by methanol extract of a the white variety stored in MAP with air, O2, CO2 and N2; and b the brown variety stored in MAP with air, O2, CO2 and N2 for 22 days at 4 °C; and Inhibition of α-glucosidase by methanol extract of c the white variety stored in MAP with air, O2, CO2 and N2; and d the brown variety stored in MAP with air, O2, CO2 and N2 for 22 days at 4 °C. Some error bars are too small to be visible

The brown variety appeared to be much more powerful inhibitor; its activity never dropped below 60% regardless of the type of the MAP sample tested (Fig. 2b).

Portobello stored in O2 and CO2 exhibited almost the same behavior as the control sample, except N2, the α-amylase inhibition activity of which was significantly lower but still high during the first two weeks of cold storage. Tamboli et al. [45] showed pronounced α-amylase inhibition activity of various extracts obtained from Pleurotus ostreatus mycelium, connecting this activity with flavonoids (especially catechin) and glycoproteins. In the present study, we did not find the dependence between the total flavonoid content and α-amylase inhibitory activity. Compounds like fatty acids are also proved as inhibitors of diabetes connected enzymes as well as glycoproteins [41]. Synergy between constituents of the button mushroom might also be present when it comes to α-amylase inhibition. However, we showed that the moment of mushroom purchase and consumption as well as the type of shelf-life extending technique had a significant impact on this functional aspect of A. bisporus.

Another enzyme important for prevention and control of diabetes mellitus, α-glucosidase, was also inhibited by the methanol extracts of A. bisporus (Fig. 2c and d). The maximal measured activity exceeded 50%, although overall activity was much lower than that of α-amylase. This is not the first case that natural compounds have different behavior against these two enzymes [41]. The white button mushroom was a particularly poor inhibitor of α-glucosidase, since activity disappeared completely between the first and the second week of storage (Fig. 2c). Almost all MAPs were non-effective in preventing the loss of high α-glucosidase inhibitory activity present at the beginning of the storage period. This indicates that A. bisporus is very sensitive in the post-harvest period and that none of the parameters tested were affected like α-glucosidase activity. Surprisingly, N2 packaging showed stable, and even stimulating effect on this parameter. Even, quite contrary to α-amylase activity. The brown variety maintained the ability to inhibit this enzyme, although it had a wave-like pattern, with a day-by-day statistically significant variation. Depending on the storage time, different MAP packaging showed the best stimulation of α-glucosidase inhibition activity in the brown strain.

Matsuura et al. reported less than 50% of α-glucosidase inhibitory ability of wild A. bisporus ethanol extract and about 30% in the case of water extract [46]. This is in accordance with our findings for the fresh sample of the white strain.

The explanation of these changes is beyond the scope of this study, requiring further research, but this phenomenon is also first time reported and expands a simple view on mushrooms as functional food. Not just that strain, genes, cultivation and storage conditions affect mushroom’s nutritional quality, but they also have significant effect on functional characteristics, as discussed in this study.


Although mushrooms do have functional characteristics, contemporary life style and high demand for extended shelf life puts under the question the functional attributes and levels of nutraceuticals in particular food. Preserving techniques (like MAP) can maintain sensory and some nutritional attributes of mushrooms. Based on our findings, MAP can efficiently save, and even promote, selected functional characteristics of A. bisporus for two weeks of cold storage. For most of the measured parameters in white strain packaging with a high level of O2 had the best effect. In case of Portobello CO2 MAP was more effective. The strongest effect on α-glucosidase inhibition activity of white strain was obtained when the mushrooms were stored in N2 packaging. The effect was stable for all 22 days of shelf life. On the contrary, no clear conclusion can be made in the case of the brown variety. On the other side, commercial packaging negatively affected mushroom’s functional characteristics in most of the examined parameters. More in-depth research is needed to elucidate if the food, such as mushrooms, can be called functional after they spend some time on a retail shelf or under changed storage conditions.


  1. 1.

    FAO Diversification booklet number 7. http://www.fao.org/3/a-i0522e.pdf (Accessed 31 March 2019)

  2. 2.

    Royse DJ, Baars J, Tan Q (2017) In: Zied DC, Pardo-Giménez (eds.) Edible and medicinal mushrooms: technology and applications. John Wiley & Sons, Hoboken

  3. 3.

    Zhang K, Pu YY, Sun DW (2018) Recent advances in quality preservation of postharvest mushrooms (Agaricus bisporus): A review. Trends Food Sci Technol 78:72–82

    CAS  Article  Google Scholar 

  4. 4.

    Singh P, Langowski LC, Wani AA, Saengerlaub S (2010) Recent advances in extending the shelf life of fresh Agaricus mushrooms: a review. J Sci Food Agric 90:1393–1402

    CAS  Article  Google Scholar 

  5. 5.

    Royse DJ (2014) A global perspective on the high five: Agaricus, Pleurotus, Lentinula, Auricularia & Flammulina. Proceedings of the 8th international conference on Mushroom biology and mushroom products (ICMBMP8) 1–6

  6. 6.

    The United States International Trade Commission, https://www.usitc.gov/publications/332/ITS_7.pdf (Accessed 6 Apr 2019)

  7. 7.

    Thangthaeng N, Miller MG, Gomes SM, Shukitt-Sale B (2015) Daily supplementation with mushroom (Agaricus bisporus) improves balance and workingmemory in aged rats. Nutr Res 35:1079–1084

    CAS  Article  Google Scholar 

  8. 8.

    Gholami R, Ahmadi E, Farris S (2017) Shelf life extension of white mushrooms (Agaricus bisporus) by low temperatures conditioning, modified atmosphere, and nanocomposite packaging material. Food Packag Shelf Life 14:88–95

    Article  Google Scholar 

  9. 9.

    Ventura-Aguilar RI, Colinas-León MT, Bautista-Bańos S (2017) Combination of sodium erythorbate and citric acid with MAP, extended storage life of sliced oyster mushrooms. LWT-Food Sci Technol 79:437–444

    CAS  Article  Google Scholar 

  10. 10.

    Lin X, Sun DW (2019) Research advances in browning of button mushroom (Agaricus bisporus): Affecting factors and controlling methods. Trends Food Sci Technol 90:63–75

    CAS  Article  Google Scholar 

  11. 11.

    Atila F, Owaid MN, Shariati MA (2017) The nutritional and medicinal benefits of Agaricus bisporus: a review. J Microbiol Biotechnol Food Sci 7:281–286

    CAS  Google Scholar 

  12. 12.

    Barros L, Falcão S, Baptista P, Freire C, Vilas-Boas M, Ferreira ICFR (2008) Antioxidant activity of Agaricus sp. mushrooms by chemical, biochemical and electrochemical assays. Food Chem 111:61–66

    CAS  Article  Google Scholar 

  13. 13.

    Vunduk J, Djekic I, Petrovic P, Tomašević I, Kozarski M, Despotović S, Nikšić M, Klaus A (2018) Challenging the difference between white and brown Agaricus bisporus mushrooms. Br Food J 120:1381–1394

    Article  Google Scholar 

  14. 14.

    Ekowati N, Yuniati NI, Hernayanti H, Ratnaningtyas NI (2018) Antidiabetic potentials of button mushroom (Agaricus bisporus) on alloxan-induced diabetic rats. Biosaintifika 10:655–662

    Article  Google Scholar 

  15. 15.

    Packialakhsmi B, Loganayagi CT (2018) Antihyperglycemic activity of Agaricus bisporus mushroom extracts on alloxan induced diabetic rats. Int J Pharm Res Health Sci 6:2475–2479

    Google Scholar 

  16. 16.

    Barros L, Ferreira MJ, Queiroś B, Ferreira ICFR, Baptista P (2007) Total phenols, ascorbic acid, β-carotene and lycopene in Portuguese wild edible mushrooms and their antioxidant activities. Food Chem 103:413–419

    CAS  Article  Google Scholar 

  17. 17.

    Novaković AR, Karaman MA, Matavulj MN, Pejin BM, Belović MM, Radusin TI, Ilić NM (2015) An insight into in vitro bioactivity of wild-growing puffball species Lycoperdon perlatum (Pers). Food Fed Res 42:51–58

    Article  Google Scholar 

  18. 18.

    Santos JS, Brizola VFA, Granato D (2017) High-throughput assay comparison and standardization for metal chelating capacity screening: A proposal and application. Food Chem 214:515–522

    CAS  Article  Google Scholar 

  19. 19.

    Kirakosyan A, Gutierrez E, Solano BR, Seymour EM, Bolling SF (2018) The inhibitory potential of Monmorency tart cherry on key enzymes relevant to type 2 diabetes and cardiovascular disease. Food Chem 252:142–146

    CAS  Article  Google Scholar 

  20. 20.

    López-Vaźquez E, Prieto-García F, Gayosso-Canales M, Otazo Sánchez EM, Villagómez Ibarra JR (2017) Phenolic acids, flavonoids, ascorbic acid, β-glucans and antioxidant activity in Mexican wild edible mushrooms. Ital J Food Sci 29:766–774

    Google Scholar 

  21. 21.

    Jaworska G, Pogoń K, Bernaś E, Duda-Chodak A (2015) Nutraceuticals and antioxidant activity of prepared for consumption commercial mushrooms Agaricus bisporus and Pleurotus ostreatus. J Food Qual 38:111–122

    CAS  Article  Google Scholar 

  22. 22.

    Smirnoff N (2018) Ascorbic acid metabolism and functions: A comparison of plant and mammals. Free Radic Biol Med 122:116–129

  23. 23.

    Georgiou CD, Zervoudakis G, Petropoulou KP (2003) Ascorbic acid might play a role in the sclerotial differentiation of Sclerotium rolfsii. Mycologia 95:308–316

    CAS  Article  Google Scholar 

  24. 24.

    Li T, Zhang M (2013) The physiological and quality change of mushroom Agaricus bisporus stored in modified atmosphere packaging with various sizes of silicone gum film window. Food Sci Technol Res 19:569–576

    CAS  Article  Google Scholar 

  25. 25.

    Ajibola VO, Babatunde OA, Suleiman S (2009) The effect of storage method on the vitamin c content in some tropical fruit juices. Trends Appl Sci Res 4:79–84

    CAS  Article  Google Scholar 

  26. 26.

    Bhat R, Stamminger R (2016) Impact of combination treatments of modified atmospherepackaging and refrigeration on the status of antioxidants in highly perishable strawberries. J Food Process Eng 39:121–131

    CAS  Article  Google Scholar 

  27. 27.

    Liu Z, Wang X, Zhu J, Wang J (2010) Effect of high oxygen modified atmosphere on post-harvest physiology and sensorial qualities of mushrooms. Int J Food Sci Tech 45:1097–1103

    CAS  Article  Google Scholar 

  28. 28.

    Palacios I, Lozano M, Moro C, D’Arrigo M, Rostagno MA, Martínez JA, García-Lafunete A, Guillamón E, Villares A (2011) Antioxidant properties of phenolic compounds occuring in edible mushrooms. Food Chem 128:674–678

    CAS  Article  Google Scholar 

  29. 29.

    Kaewnarin K, Suwannarach N, Kumla J, Lumyong S (2016) Phenolic profile of various wild edible mushroom extracts from Thailand and their antioxidant properties, anti-tyrosinase and hyperglycaemic inhibitory activities. J Funct Foods 27:352–364

    CAS  Article  Google Scholar 

  30. 30.

    Gąsecka M, Mleczek M, Siwulski M, Niedzielski P, Kozak L (2015) The effect of selenium on phenolics and flavonoids in selected edible white rot fungi. LWT-Food Sci Technol 63:726–731

    Article  Google Scholar 

  31. 31.

    Valliyodan B, Shi H, Nguyen HT (2015) A simple analytical method for high-throughput screening of major sugars from soybean by normal-phase HPLC with evaporative light scattering detection. Chromatogr Res Int. https://doi.org/10.1155/2015/757649

    Article  Google Scholar 

  32. 32.

    Estevinho BN, Ferraz A, Santos L, Rocha F, Alves A (2013) Uncertainty in the determination of glucose and sucrose in solutions with chitosan by enzymatic methods. J Braz Chem Soc 24:931–938

    CAS  Google Scholar 

  33. 33.

    Sławińska A, Jabłońska-Ryś E, Stachniuk A (2020) High-performance liquid chromatography determination of free sugars and mannitol in mushrooms using corona charged aerosol detection. Food Anal Methods. https://doi.org/10.1007/s12161-020-01863-8

    Article  Google Scholar 

  34. 34.

    Yan Z, Zhao M, Wu X, Zhang J (2020) Metabolic Response of Pleurotus ostreatus to continuous heat stress. Front Microbiol. https://doi.org/10.3389/fmicb.2019.03148

    Article  PubMed  PubMed Central  Google Scholar 

  35. 35.

    Surya S, Ajit S, Asgar A, Sivakumar M (2018) Effect of carbon dioxide on the physicochemical quality of Ganoderma lucidum. Biotech Today: Int J Biol Sci 8:7–19

    Google Scholar 

  36. 36.

    Atala E, Fuentes J, Wehrhahn MJ, Speisky H (2017) Quercetin and related flavonoids conserve their antioxidant properties despite undergoing chemical or enzymatic oxidation. Food Chem 234:479–485

    CAS  Article  Google Scholar 

  37. 37.

    Abugri DA, McElhenney WH (2013) Extraction of total phenolic and flavonoids from edible wild and cultivated medicinal mushrooms as affected by different solvents. J Nat Prod Plant Resour 3:37–42

    CAS  Google Scholar 

  38. 38.

    Shao Y, Guo H, Zhang J, Liu H, Wang K, Zuo S, Xu P, Xia Z, Zhou Q, Zhang H, Wang X, Chen A, Wang Y (2020) The genome of the medicinal macrofungus Sanghuang provides insights into the synthesis of diverse secondary metabolites. Front Microbiol. https://doi.org/10.3389/fmicb.2019.03035

    Article  PubMed  PubMed Central  Google Scholar 

  39. 39.

    Piljac-Žegarac J, Valek L, Martinez S, Belščak A (2009) Fluctuations in the phenolic content and antioxidant capacity of dark fruit juices in refrigerated storage. Food Chem 113:394–400

    Article  Google Scholar 

  40. 40.

    Djekic I, Vunduk J, Tomašević I, Kozarski M, Petrovic P, Niksic M, Pudja P, Klaus A (2017) Total quality index of Agaricus bisporus mushrooms packed in modified atmosphere. J Sci Food Agric 97:3013–3021

    CAS  Article  Google Scholar 

  41. 41.

    De Silva DD, Rapior S, Hyde KD, Bahkali AH (2012) Medicinal mushrooms in prevention and control of diabetes mellitus. Fungal Divers 56:1–29

    Article  Google Scholar 

  42. 42.

    Su CH, Lai MN, Ng LT (2013) Inhibitory effects of medicinal mushrooms on α-amylase and α-glucosidase – en. Food Funct 4:644–649

    CAS  Article  Google Scholar 

  43. 43

    Sarikurkcu C, Andrade JC, Ozer MS, de Lima Silva JMF, Ceylan O, de Sousa EO, Coutinho HDM (2020) LC-MS/MS profiles and interrelationships between the enzyme inhibition activity, total phenolic content and antioxidant potential of Micromeria nervosa extracts. FoodChem 328:126930

    CAS  Google Scholar 

  44. 44.

    Takaya N (2009) Response to hypoxia, reduction of electron receptors, and subsequent surviaval by filamentous fungi. Biosci Biotechnol Biochem 73:1–8

    CAS  Article  Google Scholar 

  45. 45.

    Tamboli E, Bhatnagar A, Mishra A (2018) Alpha-amylase inhibitors from mycelium of an oyster mushroom. Prep Biochem Biotech 48:693–699

    CAS  Article  Google Scholar 

  46. 46.

    Matsuura H, Asakawa C, Kurimoto M, Mizutani J (2002) α-glucosidase inhibitor from the seeds of balsam pear (Momordica charantia) and the fruit bodies of Grifola frondosa. Biosci Biotechnol Biochem 66:1576–1578

    CAS  Article  Google Scholar 

Download references


This study was undertaken with the financial support of the Ministry of Education and Technological Development of the Republic of Serbia (project III 46010) Grant no: Contact No. 200116. Special thanks to Ivana Bojkić, a professional translator, for proofreading the article and English language corrections.

Author information




JV: Conceptualization, Methodology, Investigation, Formal analysis, Data curation, Writing-original version and editing. MK: Methodology, Investigation, Formal analysis. IDj: Conceptualization, Data curation, Statistical analysis, Writing-review and editing. IT: Conceptualization, Writing-review and editing. AK: Methodology, Supervision, Project Administration, Writing-review and editing.

Corresponding author

Correspondence to Jovana Vunduk.

Ethics declarations

Conflict of interest

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Compliance with ethics requirements

This article does not contain any study with human or animal subjects.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Vunduk, J., Kozarski, M., Djekic, I. et al. Effect of modified atmosphere packaging on selected functional characteristics of Agaricus bisporus. Eur Food Res Technol (2021). https://doi.org/10.1007/s00217-020-03666-x

Download citation


  • α-amylase inhibition
  • α-glucosidase inhibition
  • Functional food
  • Modified atmosphere packaging
  • Shelf life