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Latin American Applied Research 52(3):221-226 (2022)
https://doi.org/10.52292/j.laar.2022.934 221
INFLUENCE OF CHITOSAN COATINGS ON THE POSTHARVEST
SHELF LIFE AND QUALITY OF BUTTON MUSHROOMS (AGARICUS
BISPORUS) DURING COLD STORAGE
E. NAKILCIOĞLU and S. ÖTLES
† Food Engng. Department, Ege Univ., Bornova, Izmir, 35100, Turkey.
emine.nakilcioglu@ege.edu.tr, semih.otles@ege.edu.tr
Abstract
−−
In this study the influence of chitosan
coating on the physiochemical properties of button
mushrooms stored at 4 ºC was investigated. The but-
ton mushrooms were coated with 1 %, 2 % and 3 %
(w/v) chitosan and then stored for 20 days. The results
showed that chitosan coating is influential in inhibit-
ing the senescence of button mushrooms and preserv-
ing their nutritional qualities. The optimum coating
rate for button mushrooms in which the respiratory
rate (28.26 %), weight loss (54.97 %) and percentage
of open caps (72.22 %) are reduced and the changes
in polyphenols (42.18 %), antioxidant capacity (4.38
%), ascorbic acid (33.29 %), titratable acidity (27.56
%) and total soluble solids (40.92 %) are retarded,
was determined to be 2 %. The findings of this study
suggest that 2 % (w/v) chitosan can be utilized for ex-
tending the postharvest shelf life of button mush-
rooms by delaying the ripening process during cold
storage.
Keywords
−−
Antioxidant capacity, ascorbic acid,
edible coating, nutritional quality, respiratory rate.
I. INTRODUCTION
Consumers all around the world demand the production
of more natural and environmentally friendly foods that
have a longer shelf life and are of high quality without
the use of any chemical preservatives. Thus, the use of
edible coatings, an environmentally friendly bio-based
technology used to extend the shelf life of foods, has be-
come a popular strategy (Gol et al., 2013). Chitosan,
which is an edible coating agent, is the second most abun-
dant polysaccharide after cellulose in nature and has at-
tracted a great deal of scientific and agricultural interest
due to its unique characteristics such as biocompatibil-
ity, film-forming and antimicrobial effect (Yu et al.,
2018). Many studies have been carried out on the use of
chitosan as an edible coating to preserve the quality of
various fruits and vegetables, however very little is
known about its effects on the quality characteristics of
button mushrooms.
Agaricus bisporus, known as the button mushroom, is
one of the most economically considerable edible mush-
rooms. It is of great nutritional value with high contents
of ergothioneine, polyphenols, minerals, vitamins and
polysaccharides. Moreover, it has several important bio-
logical activities such as antitumor, anti-inflammatory,
antimicrobial, anti-aromatase, immunomodulatory and
antioxidant properties (Liu et al., 2013). Although the
button mushroom is consumed worldwide, its commer-
cial potential is limited due to its short shelf life which is
around 8 days in refrigerated conditions (Gholami et al.,
2017). Therefore, an economic, adequate, and efficient
postharvest preservation of the mushroom to extend its
shelf life is of great necessity which would be profitable
for both the mushroom industry and the consumers.
Considering the increasing demand for fresh food,
many preservation methods have been developed includ-
ing low temperature storage, γ-irradiation, modified at-
mosphere packaging and chemical treatments in order to
slow down the postharvest deterioration rate of fresh
mushrooms (Nasiri et al., 2018). The application of a
semi-permeable edible coating such as chitosan may
have the same effect as storing the mushrooms in a mod-
ified atmosphere to prolong their shelf life. Chitosan
coatings act as a barrier to gas exchange and water vapor
transfer. They reduce the gas exchange between the
mushrooms and the surrounding atmosphere by creating
a modified interior atmosphere (low O2 and high CO2)
and decreasing water loss (Nasiri et al., 2018). The
preservation of food quality and the extension of posthar-
vest life was achieved by using chitosan coatings in man-
gos (Kittur et al., 2001), sliced button mushrooms (Eissa,
2007), Eksotika II papayas (Ali et al., 2011), guavas
(Hong et al., 2012; Sneha Nair et al., 2018), shiitake
mushrooms (Jiang et al., 2012), bananas (Wang and Gao,
2013), asparagus (Qiu et al., 2013), Cavendish bananas
(Suseno et al., 2014), red kiwifruit (Kaya et al., 2016),
pomegranate arils (Ozdemir and Gokmen, 2017), plums
(Kumar et al., 2017), table grapes (Gao et al., 2013;
Castelo Branco Melo et al., 2018) and grass carp fillets
(Yu et al., 2018). According to the literature review, the
use of chitosan coating in whole fresh button mushrooms
has never been studied before. Furthermore, no studies
investigating the influence of 3 % (w/v) chitosan coating
on the shelf life and quality of the product were found.
The aim of this study was to evaluate the effects of
edible coatings using chitosan at different concentrations
on the postharvest quality of button mushrooms stored at
4ºC for 20 days and to determine the formulation of the
chitosan coating that extends the shelf life of the mush-
rooms by best preserving their postharvest qualities. By
determining and using the proper formulation of chitosan
Latin American Applied Research 52(3):221-226 (2022)
222
coating, it was also aimed to help keep the quality char-
acteristics of the mushrooms at maximum level during
storage.
II. METHODS
A. Coating Treatments
The button mushrooms used in this study, all of which
were similar in terms of size, shape and color, were pur-
chased from commercial market. The purchased mush-
rooms had been separated from the bed one day ago. For
surface-sterilization, the mushrooms were immersed in a
0.1 % (v/v) NaClO solution during 1 min and then air-
dried at room temperature for 30 min. Chitosan solutions
of various concentrations (1.0, 2.0, 3.0 % w/v) were pre-
pared from hydrosoluble chitosan powder (chitosan from
crab shells, 91.6% deacetylation, Qingdao Reach Inter-
national Inc., Qingdao, China) in the solution containing
2 % (w/v) malic acid with a pH value of 6.0. The button
mushrooms were dipped into the chitosan solutions for 1
min. A control group was prepared from the mushrooms
treated only with 2 % (w/v) malic acid solution (Eissa,
2007). All coated samples were placed in macroperfo-
rated polypropylene film bag (40 μm thickness, 0.2mm2
surface, 1.3 × 104 perforations/m2) and stored at 4 °C for
20 days to evaluate both their postharvest shelf life and
qualities every 5 days. All of the samples were prepared
in duplicates.
B. Respiration Rate, Percentage of Open Caps and
Weight Loss
Respiration rate was performed according to the method
reported by Li et al. (2006) with various modifications.
A closed system was used to determine the respiration
rate of the mushrooms. 50 g of randomly selected mush-
rooms were placed in a Petri-dish filled with 10 mL of
0.4 M NaOH. Then, they were kept in a desiccator during
30 min. The residual NaOH in the Petri-dish was titrated
with 0.4 M oxalic acid. The respiratory rate was calcu-
lated from the consumed oxalic acid volume.
The development of an umbrella-like shape on the
cap followed by the detachment of the veil were taken as
the criteria for evaluating the percentage of open caps.
The percentage of open caps was expressed as the ratio
of the number of open capped mushrooms to the total
number of mushrooms in the package (Jiang, 2013).
Weight loss was determined in accordance with the
methods of Ares et al. (2006) by weighing the packages
of mushrooms before and after the storage period.
Weight loss was determined as the percentage of the loss
of weight concerning the initial weight.
C. Total Soluble Solids and Titratable Acidity
Twenty grams of mushrooms were homogenized and
centrifuged (multispeed centrifuge IEC CL31; Thermo
Fisher Scientific Inc., Wilmington, DE, USA) for 20 min
at 10,000 × g. The collected supernatant was then ana-
lyzed. The percent soluble solids were determined by a
digital refractometer (RFM 330; Bellingham + Stanley
Ltd, Kent, UK) and expressed as Brix (Eissa, 2007). The
device reads ºBrix values at corrected temperature levels.
The titratable acidity was also determined according to
the method described by Ali et al. (2011). Ten grams of
mushrooms were homogenized with 40 mL of distilled
water using a commercial blender (7010S; Waring Com-
mercial, Torrington, Connecticut USA). The mixture was
filtered and then 5 mL of the filtrate was titrated with 0.1
M NaOH in the presence of phenolphthalein as an indi-
cator. Titratable acidity was expressed as a percentage of
citric acid per 100 g fresh mushrooms.
D. Antioxidant Capacity, Total Polyphenol Content
and Ascorbic Acid Content
For the preparation of the polyphenol extracts, 0.5 g of
mushrooms were extracted with 10 mL of pure methanol
by centrifugation (multispeed centrifuge IEC CL31;
Thermo Fisher Scientific Inc., Wilmington, DE, USA) at
936 × g for 15 min at 4°C (Mami et al., 2014). The abil-
ities of the polyphenol extracts obtained from the mush-
rooms to scavenge 2,2-diphenyl-1-picrylhydrazyl
(DPPH) free radical were determined by modifying the
methods of Cheung et al. (2003). One mililiter of extract
or methanol (control sample) was mixed with 0.5 mL of
0.1 mM DPPH radical prepared in methanol. The mixture
was vortexed (test tube shaker TTS 2; IKA-Werke
GMBH and CO.KG, Lille, France) and the absorbance
was read immediately at 520 nm with an UV-vis spectro-
photometer (Optizen Pop; Mecasys Co., Ltd., Korea).
The results were expressed as the percentage of inhibi-
tion. The total polyphenol contents of the mushrooms
were determined using the Folin-Ciocalteu colorimetric
method (Mami et al., 2014). Three hundred microliters
of the extract were mixed with 200 μL of distilled water
and 2.5 mL of 10 % Folin-Ciocalteu reagent. After stand-
ing for 6 min, 2 mL of 7.5 % sodium carbonate solution
was added to the mixture. The mixture was kept in the
dark and at room temperature for 90 min and then its ab-
sorbance was determined by a UV/vis spectrophotometer
(Optizen Pop; Mecasys Co., Ltd., Korea) at 760 nm. The
results were evaluated as mg gallic acid equivalent per
100 g fresh mushrooms.
The ascorbic acid contents of the mushrooms were
carried out using the 2,6- dichlorophenolindophenol titra-
tion method developed by Mami et al. (2014). Six grams
of mushrooms were mixed with 40 mL of 2 % (w/v) ox-
alic acid and then homogenized. Ten mL of supernatant
was titrated by 2,6-dichlorophenolindophenol solution
and the results were expressed as mg ascorbic acid per
100 g of fresh mushrooms.
E. Statistical Analysis
All experiments were replicated twice, and their results
were reported as means ± standard deviations. One-way
analysis of variance (ANOVA) was applied on the data
sets using SPSS 20.0 statistical package program. Statis-
tical significances were determined (p<0.05). Duncan
multiple range test was used for evaluating the significant
difference amongst the samples.
E. NAKILCIOĞLU, S. ÖTLES
223
Figure 1: Effect of chitosan coating on respiration rates (A),
percent open caps (B) and weight losses (C) of button mush-
rooms stored at 4 °C during 20 days. Vertical bars represent
standard deviations of means.
III. RESULTS
A. Evaluation of Changes in Respiration Rate, Per-
centage of Open Caps and Weight Loss
The main properties of the respiration rates, percentage
of open caps and weight losses of the button mushrooms
coated with different ratios of chitosan are shown in Fig.
1. During the storage period, the respiration rates of the
mushrooms significantly decreased while their percent-
age of open caps and weight losses significantly in-
creased (p<0.05). The respiratory rates, percentage of
open caps and weight losses of the coated mushrooms
were lower than those of the control group (p<0.05). At
the beginning of the storage period, the respiratory rate
of the control group was 78.30 % while this rate was 1.26,
1.39 and 1.34 times higher than those of the 1 % chitosan,
2 % chitosan and 3 % chitosan-coated mushrooms at the
end of the storage period, respectively. Throughout the
storage period, 36.91 % reduction in weight was ob-
served in the control group while 34.63 %, 16.62 % and
26.01 % weight losses occurred in the 1 % chitosan, 2 %
chitosan and 3 % chitosan-coated mushrooms, respec-
tively. The percentage of open cap mushrooms in the
control group was 50.0 % after 20 days in storage. On the
other hand, the percentages of open caps in the mush-
rooms coated with 1 % (w/v) chitosan, 2 % (w/v) chi-
tosan and 3 % (w/v) chitosan were in the range of 13.89
(2 % chitosan-coated mushrooms) – 32.14 % (1 % chi-
tosan-coated mushrooms) after 20 days. The mushrooms
coated with 2 % (w/v) chitosan showed the lowest respir-
atory rates, percentage of open caps and weight loss
(p<0.05). The results of this study regarding the changes
in the respiration rates, percentage of open caps and
weight loss during the storage period of the mushrooms
were consistent with the previous works of Ali et al.
(2011), Jiang et al. (2012), Jiang (2013), Gao et al.
(2013), Suseno et al. (2014), Kaya et al. (2016), Kumar
et al. (2017), and Sneha Nair et al. (2018).
Chitosan coating modifies the internal atmosphere of
the mushrooms. It leads to a reduction of CO2 production
in the coated mushrooms and thereby, the respiration of
the mushrooms slows down (Jiang et al., 2012). Gas ex-
change between the mushrooms and the surrounding at-
mosphere occurs through the open pores that exist as a
result of the high permeability of the skin of the mush-
rooms. Owing to the partial blockage of the pores exist-
ing in mushrooms with chitosan coating, moisture loss is
partially prevented and the respiratory rate is kept under
control (Castelo Branco Melo et al., 2018). The cap open-
ing of the mushrooms is a result of the mushrooms drying
due to water loss during the storage period. Increased wa-
ter loss during the storage period causes a reduction in
the cohesive forces of water and molecules like proteins
responsible for the entire position of the cap and veil of
mushroom. Chitosan coating also reduces the percentage
of open cap by reducing the water loss in mushrooms
(Jiang, 2013). Due to the fact that the ripening process
continues throughout the storage period, the percentage
of open cap and water loss in mushrooms increases while
the rate of respiration decreases.
B. Evaluation of Changes in Total Soluble Solids and
Titratable Acidity
The data of total soluble solids content and titratable
acidity, which are important quality properties related to
the ripening and maturation of the button mushrooms
coated with different ratios of chitosan, are presented in
Table 1. Chitosan coating caused a significant increase in
the total soluble solids content of the mushroom
(p<0.05). The total soluble solids content exhibited an in-
creasing trend throughout the 20 day storage period
(p<0.05) while the titratable acidity did not change sig-
nificantly throughout the storage period, except for in the
control group, and was not affected by the coating appli-
cation (p>0.05).
During the 20 day storage period, the increase of total
soluble solid content was in the range of 14.63 (2 % chi-
tosan-coated mushrooms) – 21.43 % (1 % chitosan-
coated mushrooms) in the coated mushrooms while it
was 55.56 % in the control group. At the end of the stor-
Latin American Applied Research 52(3):221-226 (2022)
224
Table 1. Total soluble solids, and titratable acidity in control group and 1 % chitosan-coated mushrooms, 2 % chitosan-coated
mushrooms and 3 % chitosan-coated mushrooms during storage at 4°C.
Treatment
t(storage)/day
control
w(chitosan)/1 %
w(chitosan)/2 %
w(chitosan)/3 %
Total soluble
solids/°Brix
0
(4.05±0.00)eD
(4.20±0.01)dA
(4.10±0.00)dC
(4.15±0.01)dB
5
(5.30±0.00)dA
(4.35±0.07)cB
(4.15±0.07)cC
(4.20±0.00)cC
10
(5.45±0.07)cA
(4.60±0.00)bB
(4.40±0.00)bC
(4.55±0.07)bB
15
(5.60±0.00)bA
(4.60±0.00)bB
(4.45±0.07)bC
(4.53±0.04)bBC
20
(6.30±0.00)aA
(5.10±0.00)aB
(4.70±0.00)aD
(4.95±0.07)aC
Titratable acid-
ity/Citric acid
eq. %
0
(0.12±0.01)aA
(0.12±0.02)aA
(0.12±0.01)aA
(0.13±0.02)aA
5
(0.13±0.04)aA
(0.13±0.04)aA
(0.13±0.04)aA
(0.13±0.04)aA
10
(0.09±0.01)abA
(0.10±0.01)aA
(0.12±0.02)aA
(0.12±0.03)aA
15
(0.05±0.01)bA
(0.08±0.04)aA
(0.10±0.01)aA
(0.09±0.02)aA
20
(0.04±0.01)bA
(0.06±0.02)aA
(0.08±0.01)aA
(0.07±0.01)aA
Values in the same column indicated by different small letters for each analysis are significantly different (p<0.05).
Values in the same row indicated by different capital letters for each analysis are significantly different (p<0.05).
age period, the titratable acidity in the coated mushrooms
changed from 0.06 (1 % chitosan-coated mushrooms) to
0.08 % (2 % chitosan-coated mushrooms). In addition,
the total soluble solids/titratable acidity ratio was de-
creased with the chitosan coating. It was determined that
the 2 % chitosan-coated mushrooms had the lowest total
soluble solids/titratable acidity ratio of 58.75, while the
control group had the highest total soluble solids/titrata-
ble acidity ratio of 157.50 on the 20th day of storage.
These results were in agreement with the findings of Ali
et al. (2011), Hong et al. (2012), Jiang et al. (2012), Kaya
et al. (2016) and Kumar et al. (2017) and titratable acidity
of Ozdemir and Gokmen (2017) regarding the total solu-
ble solids content.
The reduction in the total soluble solids content of
mushrooms was due to the use of chitosan coating, which
can slow the respiration and the synthesis and utilization
of the metabolites as a result of the slower hydrolysis of
carbohydrates to sugars (Jiang, 2013). Thus, the ripening
process is retarded. The chitosan coating did not cause a
significant change in the titratable acidity of the mush-
rooms. The slight changes observed in the total acidity
during the storage period were due to the mushrooms
consuming organic acids (Castelo Branco Melo et al.,
2018). As the ripening process continued throughout the
storage period, the total soluble solids content of the
mushrooms increased while the titratable acidity de-
creased. In addition, the value of titratable acidity indi-
cates the edible quality of the product (Eissa, 2007). It
was determined that the chitosan-coated mushrooms are
suitable to be processed and used fresh.
C. Antioxidant Capacity, Total Polyphenol Content
and Ascorbic Acid Content
Changes in the antioxidant capacities, total polyphenol
and ascorbic acid contents of the button mushrooms
coated with chitosan at different ratios are shown in Fig-
ure 2A-C. The initial total polyphenol content and anti-
oxidant capacity of the mushroom were 109.73 mg/100 g
FW and 85.02 %, respectively, while the initial ascorbic
acid content was 3.06 mg/100 g FW. There was a signif-
icant difference between the chitosan-coated mushrooms
and control group in terms of antioxidant capacity, total
polyphenol and ascorbic acid contents (p<0.05). Even
though, the antioxidant capacities, polyphenols, ascorbic
acid contents of both the control group and the coated
mushrooms decreased during the storage period, the use
of chitosan coating significantly reduced the decrease in
the antioxidant capacities and loss of polyphenols and
ascorbic acid contents in the mushrooms (p<0.05). After
the 20 day storage period, the antioxidant capacity of the
control group reduced by 31.08 % while the antioxidant
capacity reduction rates of the chitosan-coated mush-
rooms were in the range of 26.70 (2 % chitosan and 3 %
chitosan-coated mushrooms) - 30.07 % (1 % chitosan-
coated mushrooms). The polyphenols retentions of the
mushrooms coated with chitosan at the end of the storage
period were 58.66 % for 1 % chitosan-coated mush-
rooms, 84.54 % for 2 % chitosan-coated mushrooms and
71.67 % for 3 % chitosan-coated mushrooms, whereas
the control groups maintained only 42.36 % of initial pol-
yphenol content. In addition, the ascorbic acid retentions
of the mushrooms coated with 1 % (w/v) chitosan, 2 %
(w/v) chitosan and 3 % (w/v) chitosan were 37.05, 62.49
and 54.34 %, respectively. Whereas, the ascorbic acid re-
tention of the control group was only 29.20 % after 20
days. Based on the above results, it can be said that chi-
tosan coating has a beneficial effect in delaying the se-
nescence in mushrooms. Similar results have been ob-
tained by Hong et al. (2012), Jiang et al. (2012), Gao et
al. (2013) regarding changes in ascorbic acid content,
Kaya et al. (2016) regarding changes in both polyphenol
and ascorbic acid contents, and Kumar et al. (2017) and
Sneha Nair et al. (2018) regarding changes in the antiox-
idant capacities, polyphenol and ascorbic acid contents of
chitosan-coated products.
The reason for the higher retention of polyphenols
and ascorbic acid in the chitosan-coated mushrooms may
be due to low oxygen penetration in the coated samples
which inhibits the activities of enzymes such as polyphe-
nol oxidase, peroxidase and catalase and thus causes a
reduction in polyphenol and ascorbic acid oxidation
(Eissa, 2007; Kumar et al., 2017).
The higher antioxidant capacity of the coated mush-
rooms is associated with the better protection of polyphe-
nols and ascorbic acid (Kaya et al., 2016). Consequently,
the chitosan coated mushrooms showed higher antioxi-
dant capacity than the control group, similar to the
E. NAKILCIOĞLU, S. ÖTLES
225
Figure 2: Effect of chitosan coating on antioxidant capacities
(A), total polyphenol contents (B) and ascorbic acid contents
(C) of button mushrooms stored at 4 °C during 20 days. Vertical
bars represent standard deviations of means.
polyphenol and ascorbic acid contents during the storage
period. The continuation of metabolic activities and mat-
uration during the storage period led to a decrease in the
antioxidant capacities and polyphenol and ascorbic acid
contents of both the control group and chitosan-coated
mushrooms.
In the present study, the shelf life and almost all post-
harvest characteristics of the mushroom that were inves-
tigated were best preserved in the 2 % chitosan-coated
mushrooms, followed by the 3 % chitosan and 1 % chi-
tosan-coated mushrooms, respectively (p<0.05). The
mushrooms with 2 % (w/v) chitosan coating seemed to
inhibit the metabolic processes the most. All the men-
tioned properties improved with the increase in chitosan
concentration, except for the 3 % (w/v) chitosan coating.
The extremely high viscosity of 3 % (w/v) chitosan
caused the drying time to be prolonged, after the solution
was applied to the surface of the mushrooms, and the oat-
ing to be more difficult. This reduced the effectiveness of
the coating and made it difficult to maintain the desired
postharvest properties during the storage period.
V. CONCLUSIONS
The results of this study showed that chitosan coating can
effectively extend the shelf life of button mushrooms and
preserve their nutritional quality. Chitosan coating can
create a protective barrier on the surface of button mush-
rooms to slow respiration, reduce weight loss and per-
centage of open caps and delay changes in total soluble
solids, ascorbic acid, titratable acidity, polyphenols and
antioxidant capacity. Hence, the use of chitosan coating
in mushrooms, especially with 2 % (w/v) chitosan as the
optimum concentration, can be considered as an applica-
tion that can be used to prolong the shelf life of mush-
rooms and to maintain their nutritional quality for up to
20 days at 4 ºC. Although the chitosan coating slightly
increases the cost and sales price of mushrooms, it can
extend the shelf life of mushrooms. Thus, it can be said
that chitosan coating can reduce product loss and contrib-
ute to the economy in the long term. The overall appear-
ance of chitosan-coated mushrooms may look fine during
storage, but the safety aspect should also be addressed. In
order to use chitosan coating in mushrooms, more studies
are needed which are evaluated the microbiological as-
pects of mushrooms during storage and supported the re-
sults of this study.
ACKNOWLEDGMENT
The authors thank Sema Duvan for her technical support.
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Received: December 10, 2021
Sent to Subject Editor: December 15, 2021
Accepted: January 3, 2022
Recommended by Subject Editor Laura Briand
