International Journal of Biological Macromolecules 106 (2018) 218–226

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International Journal of Biological Macromolecules

journal homepage: www.elsevier.com/locate/ijbiomac

Application of Tragacanth gum impregnated with Satureja

khuzistanica essential oil as a natural coating for enhancement of
postharvest quality and shelf life of button mushroom (Agaricus
bisporus)
M. Nasiri a , M. Barzegar a,∗ , M.A. Sahari a , M. Niakousari b
a
Department of Food Science and Technology, Tarbiat Modares University, P.O. Box 14115-336, Tehran, Iran
b
Department of Food Science and Technology, Shiraz University, Shiraz, Iran

a r t i c l e i n f o a b s t r a c t

Article history: The effect of Tragacanth gum (TG) coating incorporated with 100, 500 and 1000 ppm Satureja khuzistan-
Received 11 June 2017 ica essential oil (SEO) on the postharvest quality and shelf life of button mushroom (Agaricus bisporus)
Received in revised form 24 July 2017 stored at 4 ± 1 ◦ C for 16 days was investigated. Weight loss, firmness, browning index (BI), total pheno-
Accepted 1 August 2017
lics, ascorbic acid, microbial and sensory quality were measured. The results indicated that treatment
Available online 2 August 2017
with TG containing SEO (TGSEO) maintained 92.4% tissue firmness, and reduced microorganism counts,
such as yeasts and molds and Pseudomonas, compared to uncoated samples. Furthermore, mushrooms
Keywords:
treated with TGSEO coating exhibited up to 57.1% decreased in BI, significantly higher levels of total phe-
Button mushroom
Food quality
nolics (85.6%) and ascorbic acid accumulation (71.8%) than control and its efficiency was better than that
Physicochemical properties TG coating alone. Sensory evaluation demonstrated the capability of TGSEO coating for preserving the
Satureja khuzistanica essential oil quality of mushroom during the storage. The results obtained endorse that application of TGSEO coating
Tragacanth gum might be a simple and effective technique for prolonging their postharvest shelf life of mushroom by up
to 16 days.
© 2017 Elsevier B.V. All rights reserved.

1. Introduction qualities for mushrooms include browning, softening, cap devel-

Since ancient times, mushrooms have been consumed by
humans not only as a part of the normal diet but also as a deli-
cacy because they have a highly desirable taste and aroma [1]. As
much as 10,378,163 t of mushrooms were produced in the world
[2]. White bottom mushroom (Agaricus bisporus) is one of the most
popular and most widely consumed edible mushroom species in
the world [3]. There are number of reasons behind the success of
this species which include relatively inexpensive cultivation meth-
ods using organic substrates and nutritional profile: high content
of riboflavin, niacin and minerals, particularly phosphorus. More-
over, A. bisporus has been considered as functional food due to the
free radical scavenging and antioxidant activities [4,5].
The short shelf life of mushrooms, typically one to three days at
room temperature, is a serious problem in postharvest distribution
[6]. They lose their commercial value within a few days. Loss of
∗ Corresponding author.
E-mail address: mbb@modares.ac.ir (M. Barzegar).
opment, off-flavour and secondary mould growth [5]. The short
shelf-life of mushroom is an impediment to the distribution and
marketing of the fresh product. Thus, prolonging postharvest stor-
age while preserving their quality would benefit the mushroom
industry as well as consumers [7]. To slow down the rate of posthar-
vest deterioration in the fresh mushrooms, there are a number of
preservative methods that could be used, such as low temperature
storage, chemical treatments, ␥-irradiation, and modified atmo-
sphere packaging [8]. Application of semipermeable edible coating
can give the same effect as modified atmosphere storage in improv-
ing the shelf life of perishable fruits [9,10]. Edible coatings act as a
barrier, decline gas exchange between fruit and the surrounding
atmosphere, result in modified interior atmosphere (high CO2 and
low O2 ), as well as reduced water loss [11]. Preservation of fruit
quality has been achieved by using a number of edible coatings,
such as chitosan in strawberries [12], carrageenan in banana [13],
gum Arabic in tomato [14] and gum cordia in chilgoza [15].
Tragacanth gum (TG) is a natural and acidic polysaccharide
with an ancient history that mostly found in certain areas of Asia
and in the semi desert and mountainous regions of Iran, Syria,

http://dx.doi.org/10.1016/j.ijbiomac.2017.08.003
0141-8130/© 2017 Elsevier B.V. All rights reserved.
 M. Nasiri et al. / International Journal of Biological Macromolecules 106 (2018) 218–226
Turkey and other near Eastern countries [16,17]. TG is defined by
Joint FAO/WHO Expert Committee on Food Additives (JECFA) as:
“a dried exudation obtained from the stems and branches of Astra-
galus gummifer Labillardière and other Asiatic species of Astragalus
(Fam. Leguminosae)” [18]. TG biopolymer is a very complex het-
erogeneous anionic branched polysaccharide with a high molecular
weight (approximately 8.4 × 105 Da) and has been widely used as a
stabilizer, thickener, emulsifier, fat replacer and cross-linking agent
in the food systems for many years [19]. It has unique chemical and
biological properties such as non-toxicity and safe for oral intake,
biocompatibility and eco-friendliness, stability over wide pH range.
Moreover, TG has been accepted since 1961 as Generally Recog-
nized as Safe (GRAS) at the level of 0.2–1.3% and in Europe has
E-number E413 on the list of additives approved by the scientific
committee for food of the European community [20,21].
In recent years, there has been an increased interest in the use
of natural antimicrobial agents instead of chemical ones. Previous
research has shown that essential oils (EOs) and extracts of many
herbs and spices are known to have antimicrobial activity which
can be used as natural food preservatives. Tragacanth gum and
the other biopolymers can be used as a suitable carrier for natural
antimicrobial and antioxidant compounds [22,23].
The genus Satureja (Lamiaceae, subfamily Nepetoideae and tribe
Satureja) constitutes about 200 species of herbs and shrubs, often
aromatic, widely distributed in Mediterranean area, Asia and boreal
America [24]. Satureja khuzistanica, “Marzeh khuzestani” in Per-
sian, is an endemic traditional herbal medicine among the nomadic
inhabitants of southwestern of Iran including Ilam, Lorestan and
Khuzestan Provinces. It used as herbal tea for its analgesic, anti-
septic and anti-inflammatory properties, especially in toothache
problems [25,26]. S. khuzistanica has also been reported to be
antispasmodic, antidiarrhea, vasodilator, antihyperlipidemic, and
antioxidant and possess antifungal, antiviral and antimicrobial
properties [27]. In previous researches S. khuzistanica essential oil
(SEO) used as natural antioxidants in soybean oil [28], safflower oil
[29] and sunflower oil [25] and antifungal agent in liquid medium
and tomato paste [30] and strawberry fruit [24]. According to our
literature review, the use of Tragacanth gum either solitary or
accompanied by Satureja khuzistanica essential oil in fresh button
mushrooms has not been studied until now.
The aim of this study was to assess the impact of TG coating with
or without different concentrations of SEO on postharvest quality
of button mushroom during storage at 4 ◦ C for 16 days.
2. Materials and methods
2.1. Plant material
Button mushrooms used in this study were harvested from a
local farm in Shiraz, Iran. Mushrooms were picked from the same
flower and from the same area of the shelf so as to reduce possible
variations caused by cultivation and environmental conditions. The
mushrooms were transferred to the laboratory within one hour of
harvesting, and then stored in darkness at 4 ± 1 ◦ C and 90% relative
humidity (RH). The mushrooms were screened for their uniformity
in size and colour and absence of mechanical damage prior to their
final processing and packaging in high density polyethylene (HDPE)
box.
2.2. Satureja khuzistanica essential oil
SEO (having purity of at least 98% as indicated in its specifica-
tion sheet), was provided by a local commercial producer of plant
essential oils.
219
2.3. Tragacanth gum and coating treatments
The high quality ribbon type TG (food grade) used in this study
was purchased from a local herbalist store, washed with water thor-
oughly and dried in a vacuum oven at 70 ◦ C. The dried gum was
ground into fine powder for subsequent experiments. In a prelimi-
nary experiment, numerous concentrations of TG, namely, 0.2, 0.4,
0.6 and 0.75% (w/v) containing 1.0% sorbitol (as a plasticizer) were
prepared and coated on button mushrooms. To optimize concen-
tration of TG solution, colorimetric test, weight loss and phenolic
contents were evaluated on each sample. Three concentrations
of SEO (100, 500 and 1000 ppm) were dissolved in 100 mL puri-
fied water at 70 ◦ C that containing 0.6% TG and 1.0% sorbitol. The
solution was homogenized to achieve complete dispersion. Five
different treatments were applied: (1) control (water); (2) Traga-
canth gum coating (TG); (3) TG coating containing 100 ppm SEO
(TGSEO1), (4) TG containing 500 ppm (TGSEO5), (5) TG containing
1000 ppm (TGSEO10). Mushrooms were dipped into their respec-
tive solutions for 5 min. The coating treatments were selected
according to the preliminary experiments on mushrooms to assure
adherence and uniformity of the coatings. The coated and control
samples were kept over a plastic strainer for 30 min and a fan gen-
erating low-speed air was used to speed up drying. The treated
samples were packaged in 18 × 20 cm high density polyethylene
(0.04 mm thickness) containers. Finally, samples were stored for
16 days at 4 ± 1 ◦ C and 95% RH for further analysis. Fifteen replicates
were included in each treatment group, and subsequently every
4 days, three replicates from each treatment group were analyzed.
2.4. Weight loss and texture analysis of samples
Weight loss was represented as the percentage of loss of weight
concerning the initial weight. In order to estimate the weight
loss content of the packages before and after the storage period
was weighed. A penetration test was implemented on the mush-
room cap using a CT3 texture analyzer (Brookfield, US), using a
5 mm diameter cylindrical probe (TA35). Samples were penetrated
5 mm in depth. The speed of the probe was 2.0 mm s−1 during the
pretest and penetration. From the force vs. time curve, firmness
was defined as the maximum force [31].
2.5. Colour
The surface colour of mushroom caps was measured with using
digital imaging and software analysis under controlled conditions
(illumination, distance between camera and sample, camera angle
and light source). The method has a good correlation with Hunter
colorimeter [32]. A white wall box without permeability to sur-
rounding light was used. A digital camera (Canon SX220 with 12.1
Mega Pixels) placed vertically at 25 cm distance from the sam-
ples. The angle between the axis of the lens and the sample was
approximately 45◦ . The images were taken at maximum resolution
and L* (light/dark), a* (red/green) and b* (yellow/blue) values of
mushroom caps measured by filter/blur/average command in CS6
Photoshop software and compared to the ideal mushroom colour
values of L* = 97, a* = −2 and b* = 0 using E as described by the
following equation [33]:
  21
E = (L − 97)2 + (a − (−2))2 + b2 (1)
where E indicates the degree of overall colour change in compar-
ison to the colour values of an ideal mushroom.
The browning index (BI), which represents the purity of brown
colour [34], was calculated according to the following equations:
BI = [100 (x − 0.31)] /0.172 (2)
220 M. Nasiri et al. / International Journal of Biological Macromolecules 106 (2018) 218–226
Fig. 1. Effect of Tragacanth gum coating containing Satureja khuzistanica essential oil on (A) weight loss (B) firmness changes of button mushrooms stored at 4 ◦ C for 16 days.
x = (a + 1.75L∗ ) / (5.645L∗ + a∗ − 3.012b∗ )
2.6. Total phenolic content determination
Total phenolic content (TPC) was determined using the Folin-
Ciocalteu colorimetric method [35]. The contents of phenolic com-
pounds were expressed as mg gallic acid equivalent per gram fresh
weight (GAE/gfw).
2.7. Ascorbic acid content analysis
The determination of ascorbic acid (AA) was carried out as
described by Fan et al. [36] by HPLC.
2.8. Respiration rate
For measuring respiration rates of the button mushrooms, a
closed system was selected. At each storage time, approximately
50 g of mushrooms from the five groups were placed under ambi-
ent air for 1 h. Then, mushrooms were stored at 20 ◦ C for 1 h in a
closed container, which contained 15 mL 0.05 M Ba(OH)2 . After that
time, 2 drops of phenolphthalein were added, and titrated with
1.44 mol L−1 oxalate. Measurements were replicated three times.
Respiration rates of the samples were expressed as CO2 production
rate and calculated using the following formula:
RI = ((V1 − V2 ) × c × 44)/(W × t)
(3) where V1 is the volume of oxalate of control (mL); V2 is the vol-
ume of oxalate of the sample (mL); c is the concentration of oxalate
(mol L−1 ); 44 is the molecular weight of CO2 ; W is the weight of the
sample (g); t is the test time (h) [37].
2.9. Microbiological analysis
All samples were analyzed for mesophilic, psychrophilic, Pseu-
domonas, and yeasts and moulds bacteria count [31]. Twenty-five
grams of mushrooms were removed aseptically from each pack
then mashed and mixed with 225 mL 0.1% peptone water. The sam-
ples were homogenized by a stomacher at high speed for 2 min.
Serial dilutions (10−1 –10−9 ) were made in serial dilution tubes by
taking 1.0 mL with 9.0 mL of 0.1% peptone water. Aerobic counts
were determined on plate count agar (PCA; Merck) following incu-
bation at 35 ◦ C for 2 days for mesophilic bacteria, and at 4 ◦ C for
7 days for psychrophilic bacteria. Pseudomonas was counted on
cephaloridine fucidin cetrimide agar (CFC; Difco), with selected
supplement SR 103 (Oxoid). The incubation temperature was 25 ◦ C
and plates were examined after 48 h. Yeasts and moulds were
estimated on potato dextrose agar (PDA; Merck) and incubation
conditions were 28 ± 1 ◦ C for 5–7 days.
2.10. Sensory evaluation
The consumer acceptance of the product was studied by 10
(4) trained panelists aged 25–40 years old. The training of the pan-
 M. Nasiri et al. / International Journal of Biological Macromolecules 106 (2018) 218–226 221

Table 1
Effect of Tragacanth gum coating containing S. khuzistanica essential oil on L* value, browning index (BI) and colour changes (E) of samples stored at 4 ◦ C for 16 days.*.

Day Control TG TGSEO1 TGSEO5 TGSEO10

L*
0 78.5 ± 0.5A,a 78.5 ± 0.5A,a 78.5 ± 0.5A,a 79.0 ± 2.0A,a 78.5 ± 0.5A,a
4 73.5 ± 0.5A,b 74.0 ± 1.0A,b 73.0 ± 1.0A,b 74.0 ± 0.0A,b 74.0 ± 1.0A,b
8 69.0 ± 0.0C,c 73.0 ± 0.0A,b 71.5 ± 0.5B,c 73.5 ± 0.5A,b 72.0 ± 0.0B,c
12 60.5 ± 1.5C,d 68.5 ± 0.5B,c 68.5 ± 0.5B,d 71.0 ± 0.0A,c 68.0 ± 0.0B,d
16 56.5 ± 1.5C,e 66.0 ± 1.0B,d 67.0 ± 0.0B,e 70.0 ± 0.0A,c 66.5 ± 0.5B,e
BI
0 4.2 ± 1.0A,d 4.9 ± 1.9A,c 2.9 ± 0.9A,e 3.6 ± 0.7A,d 4.0 ± 0.7A,d
4 12.6 ± 4.4A,c 9.0 ± 0.3A,bc 8.2 ± 1.9A,d 8.1 ± 2.1A,c 7.3 ± 0.9A,c
8 21.2 ± 1.3A,b 13.0 ± 1.8B,b 13.5 ± 2.4B,c 12.1 ± 1.6B,b 11.9 ± 2.2B,b
12 35.6 ± 4.1A,a 22.3 ± 2.8B,a 22.7 ± 0.2B,b 15.8 ± 0.8C,a 23.4 ± 1.3B,a
16 42.6 ± 0.9A,a 25.8 ± 1.5B,a 27.1 ± 0.6B,a 18.2 ± 0.0C,a 25.6 ± 0.5B,a
E
0 19.3 ± 0.9A,e 19.5 ± 1.1A,c 19.1 ± 0.5A,d 18.7 ± 3.2A,d 19.2 ± 0.7A,e
4 25.4 ± 0.9A,d 24.8 ± 1.3A,b 25.3 ± 0.9A,c 24.4 ± 0.5A,c 25.2 ± 0.2A,d
8 32.0 ± 0.4A,c 26.7 ± 0.3B,b 29.2 ± 2.4AB,b 26.0 ± 1.3B,bc 27.2 ± 0.8B,c
12 41.4 ± 2.2A,b 32.7 ± 1.3B,a 33.0 ± 0.2B,a 29.1 ± 0.0C,ab 33.4 ± 0.3B,b
16 45.4 ± 1.6A,a 35.4 ± 1.3B,a 35.3 ± 0.4B,a 30.4 ± 0.0C,a 35.0 ± 0.6B,a
*
Mean of three replicates ± standard deviation. Means in the same row with different capital letters are significantly different (p < 0.05). Means in the same column (for each
parameter) with different lowercase letters are significantly different (p < 0.05).

elists was accomplished according to the method described by Ares
et al. [38]. These attributes were: off-odour, gill colour, gill unifor-
mity, cap surface uniformity, and presence of dark zones on the cap.
Mushrooms were served in closed, odourless plastic containers at
room temperature. After opening high density polyethylene box,
mushrooms were placed in plastic containers for sensory evalua-
tion. This test was carried out within 2 h of box opening in order
to avoid loss of off-odours. A balanced complete block design was
carried out for duplicate evaluation of the samples. For scoring,
10 cm unstructured scales anchored with “nil” for zero and “high”
for ten were used, except for the gill colour descriptor, for which
the anchors were “white” and “brown”.
2.11. Statistical analysis
Experiments were conducted in triplicates. Data was analyzed
using statistical package for social scientists (SPSS version 16, IBM,
USA). Analysis of variance (ANOVA) followed by Duncan multiple
range test was used to distinguish the treatments at p < 0.05.
3. Results and discussion
3.1. Weight loss and texture of samples
During 16 days of storage, all samples experienced some weight
loss. At the end of 16 days of storage at 4 ◦ C, the control samples
showed a weight loss of 4.2% which was the highest (Fig. 1A).
The weight loss of 2.2, 2.5, 2.6 and 2.4% were recorded for
TGSEO5, TGSEO1, TGSEO10 and TG coated mushrooms, respec-
tively. Although, there was no significant difference between all
coated samples (TGSEO1, TGSEO5, TGSEO10 and TG) (p < 0.05),
but they exhibited a significantly lower weight loss in compari-
son to the control during storage (Fig. 1A). The higher weight loss
of control is attributed to mushroom’s unprotected thin epider-
mal structure resulting in a swift dehydration from surface. Lower
weight loss of coated mushrooms may be due to the characteristics
of coating as a semi-permeable barrier against O2 , CO2 , moisture
and solute movement. This is in turn reduces the respiration rate,
water loss and oxidation reaction rates [14]. TG as edible coatings
has similar function as conventional packaging, delayed mushroom
senescence and quality deterioration by reducing moisture and gas
transfer and lowering respiration [39]. Similar prevalent effects of
the edible coating treatments on weight loss have been observed in
other fruits, like alginate and gellan based coatings for on fresh-cut
Fuji apple [40] and chitosan-based coating on ‘Yali’ pears [41].
Loss of firmness influenced the mushroom quality during mar-
keting. Changes in values of firmness of control and treated
mushrooms during 16 days of storage at 4 ◦ C is illustrated in Fig. 1,
B. The initial value of 17.2 ± 0.1 N was similar for control and treated
samples (p < 0.05), and during cold storage firmness diminution
was observed for all treatments. At the end of storage period,
control mushrooms had the fastest loss of firmness, about 22.2
compared with 7.6, 8.3, 10.0, and 12.2% softening of TGSEO10,
TGSEO5, TGSEO1 and TG coated mushrooms, respectively. Soften-
ing of fruit is due to deterioration in the cell wall strength and
cellular turgor through (i) biochemical [42] and/or (ii) microbial
process [43]. Biochemical procedure involves the hydrolysis of
pectin and starch by enzymes e.g. wall hydrolases. As the pro-
cess of fruit ripening progresses, depolymerisation or shortening
of chain length of pectin substances occurs with an increase in
pectinesterase and polygalacturonase activities. Low levels of O2
and high levels of CO2 provided by TG coating may be limit the activ-
ities of these enzymes and allow retention of the firmness during
storage [44]. Concurrently, microorganisms such as Pseudomonas
destroy mushrooms by breaking down the intracellular matrix and
reducing the central vacuole, causing partially collapsed cells and
a loss of turgor in untreated samples. The kind of microorganisms
responsible for mushroom softening was detected in control sam-
ples but not in TGSEO treated mushrooms. The inhibition was due
to TGSEO higher antifungal activity and covering of the cuticle and
lenticels, thereby reducing infection, respiration and other ripen-
ing processes during storage. The results are in good agreement
with those published by Mohammadi et al. [45] who reported the
reduced transpiration and the water retention provide turgor to
the fruit cells after application with chitosan coating containing
essential oils.
3.2. Colour
Colour is the most significant factor since it is first perceived
by clients and discolouration declines the commercial worth [46].
The colour parameters lightness (L*), total colour variation (E)
and browning index (BI) which exhibited changes in the external
colourare shown in Table 1. The L* parameter is a sign of mushroom
darkening. Progressive decrease in lightness (L*) was observed dur-
ing storage in both coated and uncoated mushroom. Although, no
differences were observed between L* value of mushrooms with
222 M. Nasiri et al. / International Journal of Biological Macromolecules 106 (2018) 218–226

Fig. 2. Effect of Tragacanth gum coating containing Satureja khuzistanica essential oil on total phenolic content (mg GAE/gfw) changes of button mushrooms stored at 4 ◦ C
for 16 days.

Fig. 3. Effect of Tragacanth gum coating containing Satureja khuzistanica essential oil on ascorbic acid (mg/kg) changes of button mushrooms stored at 4 ◦ C for 16 days.

and without TG coating after 4 days, a positive influence of TG was
observed after 16 days. While, the L* value of control decreased
from 78.5 to 56.5 in 16 days, the mushroom coated with TGSEO5
showed the highest L* value (about 70.0), i.e. they generally look
whiter than the other samples after 16 days. The results were in
agreement with Eissa [10] findings for coating of mushroom with
chitosan. A semipermeable coating of TG all around the mushroom
outer surface reduces the enzymatic browning process through
restricted oxygen uptake. During storage time, since E values of
control mushrooms increased, they went darker compared to the
treated samples (Table 1). The BI values of mushrooms on the 16th
day were higher in the control sample (42.6 ± 0.9) than TGSEO5,
TGSEO10, TG and TGSEO1 groups which were 18.2 ± 0.0, 25.6 ± 0.5,
25.8 ± 1.5 and 27.1 ± 0.6, respectively.
In this study, coating of mushrooms with TG delayed colour
change, demonstrating the ability of TG coating to work as an effec-
tive barrier to oxygen, a gas necessary for browning reactions to
proceed. The present findings are similar trends as reported by
Ali et al., [14] and Khaliq et al., [11] in study of the influence of
coating on tomato and mango, respectively. In a study by Eissa
[10] for coated fresh-cut mushroom with chitosan; he concluded
that water-binding capacity of polysaccharide may repress the drip
loss of mushroom resulting in increased transparency and lowering
of lightness. Compared with control and samples coated with TG,
preservative effect may be explained by the presence of SEO. In the
same way, using EOs reduced colour changes in button mushroom
[31]. Although they presented “there is no evidence of the role of
EOs as natural compounds on colour preservation”; but SEO’s rec-
ognized antioxidant (Carvacrol) may reduce development of brown
polymers responsible for the mushrooms browning and delaying
the ripening. Therefore, synergistic effect of TG and SEO caused sig-
nificantly lower colour degradation for those samples coated with
TG coating containing SEO as compared to the controls.
3.3. Total phenolic content
As shown in Fig. 2 during the 16 days of storage, TPC decreased in
all treatments. Compared with control that showed 52.7% retention
of TPC at the end of 16th day, mushrooms coated with TGSEO5,
TGSEO10, TGSEO1 and TG preserved 85.6, 84.8, 84.0 and 78.1% of
TPC, respectively. Lower TPC in the control mushrooms might be
described by the contribution of polyphenols in the brown pigment
synthesis during storage. It seems that higher TPC correlated fairly
well with lower levels of browning, which seems to be the limiting
factor of the discolouration process. While, there was no significant
difference (p < 0.05) between TGSEO5 and TGSEO10 samples, TG
coatings enriched with SEO maintains fairly well the contents of
phenolic compounds in mushroom. In the same way, the content
 M. Nasiri et al. / International Journal of Biological Macromolecules 106 (2018) 218–226 223

Fig. 4. Effect of Tragacanth gum coating containing Satureja khuzistanica essential oil on respiration rate (mg CO2 /kg·h) of button mushrooms stored at 4 ◦ C for 16 days.

Fig. 5. Effect of Tragacanth gum coating containing Satureja khuzistanica essential oil on mesophilic (A), psychrophilic (B), Pseudomonas (C) and yeasts and moulds (D) counts
(log10 cfu/g) change of button mushrooms stored at 4 ◦ C for 16 days.

of phenolic compounds were higher than pure chitosan coatings or
uncoated controls, when coated fruits and vegetables with chitosan
containing EOs [47,48]. It is possible that essential oils would act as
‘signalling compounds’ that induce antioxidant increases in tissues
[49].
3.4. Ascorbic acid content
Ascorbic acid is water soluble and powerful antioxidants that
inhibit or decrease the damage caused by ROS in fruit and vegetable.
Changes in the ascorbic acid (AA) content of button mushrooms
224 M. Nasiri et al. / International Journal of Biological Macromolecules 106 (2018) 218–226

coated with TG and TGSEO during 16 days storage showed in Table 2

Effect of Tragacanth gum coating containing S. khuzistanica essential oil on sensory
Fig. 3. The initial ascorbic acid content of untreated button mush-
attributes change of samples stored at 4 ◦ C for 16 days.*.
rooms was 35.4 ± 0.1 ppm. Ascorbic acid degenerated progressively
in both coated and uncoated mushrooms during storage period. Day Control TG TGSEO1 TGSEO5 TGSEO10
Compared to the control sample, TGSEO10 significantly (p < 0.05) Gill colour
prevented ascorbic acid loss. The retention of ascorbic acid con- 4 1.7 ± 0.9a 1.5 ± 0.8a 1.8 ± 0.4a 1.6 ± 0.8a 1.5 ± 0.5a
tents, after 16 days of storage, for control samples and coated with 8 3.2 ± 0.6a 1.8 ± 0.4b 2.3 ± 0.8b 1.8 ± 0.8b 2.2 ± 0.8b
12 6.0 ± 1.0a 3.3 ± 0.8b 3.3 ± 0.7b 2.6 ± 0.7b 3.1 ± 0.9b
TG, TGSEO1, TGSEO5 and TGSEO10 were 40.9, 66.2, 67.8, 69.6 and
16 7.3 ± 0.8a 4.2 ± 0.8b 3.8 ± 0.8b 3.0 ± 0.8c 4.0 ± 0.9b
71.8%, respectively. Since presence of O2 can motivate ascorbic acid Dark zone
loss; the incorporation of TG to coating preparation may reduce 4 1.0 ± 0.5a 0.5 ± 0.5ab 0.6 ± 0.5ab 0.5 ± 0.5ab 0.4 ± 0.5b
O2 diffusion, slow down the respiration rate and subsequently 8 1.9 ± 0.3a 0.9 ± 0.3b 1.2 ± 0.4b 1.0 ± 0.5b 0.9 ± 0.3b
12 3.6 ± 0.8a 2.0 ± 0.5b 2.2 ± 0.4b 1.4 ± 0.5c 2.2 ± 0.4b
improve ascorbic acid content preservation, therefore delay mush-
16 4.4 ± 0.5a 2.4 ± 0.7b 2.7 ± 0.5b 1.7 ± 0.5c 2.4 ± 0.5b
room shrivelling. Similarly, Ayranci and Tunc [50] reported that Off-odour
methylcellulose-based edible coating reduced ascorbic acid loss in 4 1.8 ± 1.5a 1.2 ± 1.1a 1.3 ± 0.8a 1.2 ± 0.4a 1.4 ± 1.1a
both button mushrooms and cauliflower. The TG coating, coupled 8 3.1 ± 1.2a 2.2 ± 0.9a 3.2 ± 1.5a 2.2 ± 1.2a 2.5 ± 1.5a
with the SEO treatment, synergistically caused a higher retention 12 5.0 ± 2.0a 3.7 ± 1.4ab 3.6 ± 1.6ab 3.4 ± 1.1b 3.8 ± 1.1ab
16 6.5 ± 1.4a 4.4 ± 0.8b 3.9 ± 1.3b 3.7 ± 1.1b 4.7 ± 1.3b
of ascorbic acid in mushroom. Yuan et al., [51] stated that the abil-
Gill uniformity
ity of chitosan coating in retaining ascorbic acid content enhances 4 8.3 ± 0.5a 8.7 ± 0.5a 8.5 ± 0.5a 8.8 ± 0.4a 8.7 ± 0.7a
significantly when it is enriched with essential oils. 8 6.6 ± 0.5b 7.4 ± 0.5a 7.3 ± 0.5a 7.7 ± 0.8a 7.6 ± 0.5a
12 5.2 ± 0.8b 7.0 ± 0.7a 6.6 ± 0.5a 7.0 ± 0.7a 6.7 ± 0.7a
16 3.8 ± 0.6b 6.5 ± 0.8a 6.4 ± 0.5a 6.8 ± 0.8a 6.8 ± 0.6a
3.5. Respiration rate
Cap uniformity
4 8.5 ± 0.8ab 9.0 ± 0.0a 8.4 ± 0.5b 8.7 ± 0.5ab 8.7 ± 0.7ab
The respiration rate of button mushrooms during the 16 day 8 6.9 ± 0.6b 7.8 ± 0.6a 7.6 ± 0.5a 7.7 ± 0.5a 7.6 ± 0.8a
of storage is shown in Fig. 4. The initial respiratory rate mea- 12 5.2 ± 0.8b 6.7 ± 0.5a 6.4 ± 0.5a 6.8 ± 0.9a 6.3 ± 0.5a
sured as CO2 accumulation of mushrooms at harvest was about 16 3.9 ± 0.6b 6.1 ± 0.6a 6.1 ± 0.3a 6.3 ± 0.5a 5.9 ± 0.6a

117.33 mg CO2 kg−1 h−1 . A significant (p < 0.05) decrease in respira- *

tion rate with storage time was found for mushrooms, which could
be ascribed to mushroom decay. The respiration rate of the control
(92.39 mg CO2 kg−1 h−1 ) was higher than mushroom subjected to
TGSEO1 (78.81 mg CO2 kg−1 h−1 ), TGSEO5 (74.15 mg CO2 kg−1 h−1 ),
TGSEO10 (76.01 mg CO2 kg−1 h−1 ), and TG (79.04 mg CO2 kg−1 h−1 )
coating at the end of storage period. No significant difference was
found in the respiration rate of mushrooms coated with TG con-
taining SEO. Respiration includes energy-rich organic molecules in
cells (such as starch, sugar, and organic acids) oxidation to simpler
molecules (CO2 and H2 O), with the concurrent energy (in the form
of ATP and heat) and other molecules production [52]. Previous
researches indicate that essential oils treatments tended to have
the effective role in reducing the rate of respiration of nectarine
fruits [53], Tabarzeh grape [54] and Crimson seedless grape [55].
Moreover, apple piece treated by various bilayer coatings included
alginate presented a reduction in the rate of CO2 production [56].
The reduction in respiration rate by TGSEO treated mushrooms dur-
ing cold storage could be as a result of internal gas atmosphere
modification, which caused CO2 production reduced by coating. The
gas barrier specifications and permselectivity of the edible coat-
ing applied to the skin surface and their dependence on relative
humidity and temperature will play a key role in the changes in
inner O2 and CO2 levels. It is well known that excessive restriction
of gas exchange can lead to anaerobiosis and the development of
off-flavour [47]. On the other hand, researchers have revealed that
the essential oils isolated from Satureja could exhibit antifungal
effect [23,24]. Consequently, TGSEO could reduce respiration rate
without expansion of off-flavour.
3.6. Microbiological analysis
Antimicrobial effects of TG coating integrated with SEO
against mesophilic, psychrophilic, Pseudomonas, yeasts and moulds
revealed in Fig. 5(A–D). At the beginning, mesophilic, psychrophilic,
Pseudomonas, yeasts and moulds count of button mushroom were
4.30, 3.35, 4.82 and 3.31 log cfu/g, respectively. Incorporation of SEO
significantly enhanced the antimicrobial efficacy of TG coating. At
the end of storage time, the results presented significant inhibitory
properties of TG incorporated with SEO on both mesophilic and
psychrophilic bacteria. The impact intensifies as concentration of
Mean of three replications ± standard deviation. Means in the same row with
different letters are significantly different (p < 0.05).
SEO in TG rises. Moreover, increasing the concentration of SEO
led to a decreasing trend in the growth of yeast and moulds. The
consequences of this study indicated that SEO could reveal potent
inhibitory effects against Gram-negative, psychrotrophic bacteria,
particularly belonging to the Pseudomonasae family the main dete-
riorating bacteria found on mushroom because of contamination
of the product from compost. In our study, we investigated TG con-
taining SEO demonstrated higher antimicrobial activity compared
to TG in coating mushrooms during storage. Similarly, adding EOs
to chitosan heighten the antibacterial efficacy of chitosan coating
[51]. Jiang et al. [47] observed a synergistic effect when coated shi-
take mushroom with thyme enriched chitosan. They stated that
the polysaccharide used in the coating reduces the oil vapour dif-
fusion, in turn less loss of the EOs, rendering it more effective in
inhibiting microbial growth and maintaining the quality of mush-
rooms. Researchers have revealed that the essential oils isolated
from Satureja could exhibit antifungal effect [23,24]. The antimi-
crobial effect of Satureja khuzistanica could be due to the high
contents of phenolic compounds such as major monoterpene con-
stituents (thymol and carvacrol). Omidbeygi et al. [30] stated that
antimicrobial activity of essential oils on yeasts could be the effect
of disruption in several enzymatic systems involved in energy
production and structural components synthesis. Once the phe-
nolic compound crossed the cellular membrane, interaction with
membrane enzymes and proteins would cause an opposite flow
of protons, affecting cellular activity. Other mechanisms consist of
disturbing the functional integrity of mitochondria, preventing ATP
synthesis in the mitochondria of fungi strains and reactive oxygen
species (ROS) accumulation after using essential [24].
3.7. Sensory evaluation
Sensory evaluation of coated and uncoated mushroom during
the storage period discovered significant (p < 0.05) differences in
mushrooms gills colour, dark zones, off-odour, gill uniformity and
cap uniformity (Table 2). Developing off-odour was substantial in
the control sample after 8 days of storage. Mushroom colour pro-
 M. Nasiri et al. / International Journal of Biological Macromolecules 106 (2018) 218–226
gressively became browner during storage of all treatments. The
gills of control mushrooms showed a colour intensity of 7.3 and uni-
formity of 3.8 on the 16th day of storage. TGSEO5 coated mushroom
had the highest scores in postponement of dark spots formation
and uniformity conversion of the cap surface after 16 days of stor-
age. While there were no significant analytical differences between
TGSEOs and TG, results exhibit that TGSEO5 treatment was more
successful in mushroom sensory analysis. According to Jiang et al.
[47] the browning of mushrooms is attributed to the action of
polyphenol oxidase (PPO) enzyme and the action of bacteria and
mould on the mushroom tissue. As TG and TGSEO coatings caused
evaluation of spoilage organisms, such as Pseudomonas, respon-
sible for phenolic compounds oxidation to form brown-coloured
melanins, it inhibits brown spots development, therefore amelio-
rating the appearance and colour.
4. Conclusion
As the food industry tends to decrease the use of chemical
preservatives in food products, TG coating incorporated with EO
of S. khuzistanica could be considered as a potential source for
senescence inhibition of cold-stored mushroom. TGSEO treatment
involved the maintenance of tissue firmness and sensory quality,
reduction of microbial counts, and lowering decomposition rates of
phenolic compounds and ascorbic acid. Also, coating with TG + SEO
could retain samples’ colour compared with TG coated and con-
trol mushrooms. Furthermore, TGSEO5 showed greater influence
to inhibit discolouration or off-flavour in the button mushrooms.
The results suggest that TGSEO coating has the potential to act as
an edible coating to prolonging the button mushroom postharvest
shelf life.
Conflict of interest
The authors declare that they have no conflict of interest.
Acknowledgment
The authors acknowledge the financial support provided by the
Tarbiat Modares University Research Council.
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