Food Bioscience 37 (2020) 100722

Contents lists available at ScienceDirect

Food Bioscience
journal homepage: www.elsevier.com/locate/fbio

An ecofriendly edible coating using cashew gum polysaccharide and

polyvinyl alcohol
Bruna R. Moreira a, 1, Marcos A. Pereira-Júnior a, 1, Kátia F. Fernandes a, **, Karla A. Batista a, b, *
a
Departamento de Bioquímica e Biologia Molecular, Laboratório de Química de Polímeros, Universidade Federal de Goiás, Goiânia, Goiás, 74001-970, Brazil
b
Departamento de Áreas Acadêmicas, Instituto Federal de Educação, Ciência e Tecnologia de Goiás, Campus Goiânia Oeste, Goiânia, Goiás, 74270-040, Brazil

A R T I C L E I N F O A B S T R A C T

Keywords: Interest in food safety and plastic waste reduction has led to an increase in the search for biopolymer-based
Anacardium occidentale coatings within the food industry. The feasibility of preparing an edible coating based on cashew gum poly­
Biodegradable packaging saccharide (CGP) and polyvinyl alcohol (PVA) was studied. A CGP/PVA coating was characterized for structural,
Strawberry
mechanical, barrier and optical properties. Additionally, CGP/PVA edible coatings were bioactivated by
Fragaria ananassa
immobilization in the film of cell wall degrading enzymes (TCWDE) and applied to the surface of fresh straw­
berries for evaluation of its fungal growth inhibition capabilities. Scanning electron microscopy results showed
that the structure of the CGP/PVA coating was homogeneous without rough surfaces or pores. The CGP/PVA
edible coating showed 88% light transmission, a tensile strength of 0.04 MPa, 147% elongation at break and
water vapor permeability of 7 x 10− 11 g m− 1s− 1Pa− 1. Additionally, it was shown that CGP/PVA and CGP/PVA/
TCWDE coatings reduced strawberry-related weight loss and inhibited fungal proliferation. These results sug­
gested that the CGP/PVA hydrogel is a promising material for production of edible antimicrobial coatings.

1. Introduction
The constant need to improve food quality and safety, and the
concomitant concerns about excessive plastic waste production has
attracted interest in the development of environmentally friendly
packaging that has a low environmental impact throughout its life cycle
(Dehghani et al., 2018; Ganesan et al., 2018). Bio-based packaging
materials have undergone growth and have generated an increasing
interest in development of new materials as alternative solutions to
environmental problems caused by the accumulation of nondegradable
synthetic materials.
Effort has been made to design bio-based packaging based on natural
polymers instead of synthetic plastic materials (Bonilla & Sobral, 2016;
Hassan et al., 2018; Klangmuang & Sothornvit, 2018). Natural
polymer-based edible coatings may enhance food quality and prolong
shelf-life of food products by acting as semi-permeable membranes, thus
reducing the rate of respiration and water loss from fresh food (Gokkurt
et al., 2012). Edible coatings are defined as a thin layer of material used
for wrapping foods that may safely consumed (Dehghani et al., 2018).
These edible coatings can also reduce the rate of physiological post­
harvest degradation (Khalifa et al., 2016; Murmu & Mishra, 2018; Saberi
et al., 2018).
Among the biodegradable polymers, polyvinyl alcohol (PVA) and
cashew gum polysaccharide (CGP) have advantages over other materials
(Ma et al., 2017; Maria et al., 2008; Moreira et al., 2015; Silva et al.,
2012, 2016; Tripathi et al., 2009; Yu et al., 2018). CGP is a non-allergic,
biocompatible, and biodegradable polymer found in the gummy exudate
obtained from the bark of the cashew tree (Anacardium occidentale), that
is composed of branched heteropolysaccharides (Nayak et al., 2019;
Paula & Rodrigues, 1995).
Cashew gum extraction is ecofriendly, using a process similar to
rubber latex extraction (Nayak et al., 2019). The extracted gum is pu­
rified in an ecofriendly process to obtain the polysaccharide fraction,
using water solubilization and alcoholic precipitation. Therefore, CGP is
considered a sustainable alternative for use as a thickening and gelling
compound, colloidal stabilizer, surfactant and coating agent (Mothé

* Corresponding author. Laboratório de Química de Polímeros, ICB 2, sala 215, Universidade Federal de Goiás, Avenida Esperança, s/n, Campus Samambaia,
Goiânia, GO, CEP 74690-900, Brazil.
** Corresponding author. Laboratório de Química de Polímeros, ICB 2, sala 215, Universidade Federal de Goiás, Avenida Esperança, s/n, Campus Samambaia,
Goiânia, GO, CEP 74690-900, Brazil.
E-mail addresses: kfernandes.lqp@gmail.com (K.F. Fernandes), karla.batista@ifg.edu.br, karla-batista@hotmail.com (K.A. Batista).
1
These authors contributed equally to this work.

https://doi.org/10.1016/j.fbio.2020.100722
Received 1 July 2019; Received in revised form 24 July 2020; Accepted 25 July 2020
Available online 5 August 2020
2212-4292/© 2020 Elsevier Ltd. All rights reserved.
B.R. Moreira et al.
et al., 2017; Pitombeira et al., 2015; Ribeiro et al., 2016). However, pure
CGP films have poor mechanical properties and low stability, which
limits the use of this polymer as the only material in film formulations.
Blends of CGP with other biopolymers have been studied as re­
placements for non-sustainable synthetic materials (Cruz et al., 2019;
Oliveira et al., 2018; Rodrigues et al., 2014).
Polyvinyl alcohol (PVA) is an edible, non-toxic, film-forming and
easily prepared environment-friendly polymer that shows good chemi­
cal resistance and mechanical properties (Sanders, 1960; Silva et al.,
2013; Zanela et al., 2018). This polymer is approved by the European
Medicine Agency (EMA) and the United States Food and Drug Admin­
istration (FDA) for human use. It can be used as a component of coatings
and packaging in food applications (Curley et al., 2014; Gómez-Aldapa
et al., 2020). Incorporation of PVA into CGP-based films has been shown
to improve mechanical, morphological and physicochemical perfor­
mance of the resultant materials (Cruz et al., 2019; Silva et al., 2012,
2013). A biodegradable CGP/PVA food packaging was produced by
blending CGP and PVA that had been oxidized with sodium metaper­
iodate (Silva et al., 2012). Despite their biocompatibility and biode­
gradability, these CGP/PVA films using metaperiodate as an oxidizing
agent are not appropriate nor approved for edible coatings since the
residual iodine can affect human metabolism, leading to thyroid-related
autoimmune diseases (Knobel & Medeiros-Neto, 2004).
Therefore, the present study developed an edible CGP/PVA-based
coating using hydrogen peroxide as the oxidant agent. Hydrogen
peroxide is an unstable oxidizing agent found in biological systems,
including the human body, that slowly decomposes into non-toxic
compounds. It is a food additive that is considered safe by the Food
and Agricultural Organization (FAO)/World Health Organization
(WHO) Joint Expert Committee on Food Additives (JECFA, 2005) and is
used as an antimicrobial, preservative and sterilizing agent. Therefore,
CGP/PVA edible coatings were characterized with respect to their vis­
cosity, optical, mechanical and barrier-forming properties. This study
also evaluated the efficiency of CGP/PVA edible coatings to act as an
active covering to retard or prevent fungal growth in strawberries dur­
ing storage.
2. Materials and methods
2.1. Materials
Ethanol (95%) was purchased from Dinâmica Química Con­
temporânea Ltda (São Paulo, SP, Brazil). Hydrogen peroxide, phos­
phoric acid, sodium chloride (99% purity), calcium chloride (99%
purity), calcium nitrate (99% purity), potassium carbonate (99% pu­
rity), urea (99% purity), potassium phosphate (99% purity), ammonium
sulfate (95% purity), magnesium sulfate (99% purity), and trace ele­
ments solution (Fe2+, Mn2+, Zn2+, Co2+) were purchased from Labsynth
(São Paulo, SP, Brazil). Mannitol (98% purity), potassium tetraborate
(99% purity), sodium acetate buffer solution (50 mmol L− 1, pH 5.2),
glucose (99% purity) and potassium bromide (FTIR grade) were pur­
chased from Sigma-Aldrich Co. (São Paulo, SP, Brazil). Peptone was
purchased from Kasvi (São José dos Pinhais, PR, Brazil).).
2.2. Extraction of cashew gum polysaccharide (CGP)
Crude samples of exudate were manually collected from native
Anacardium occidentale trees between June and September 2018 in
Aquiráz, Ceará, Brazil. The exudates were stored and transported within
12 h in polystyrene cooler boxes at 20 ◦ C. The cashew gum was milled in
a Tecnal mill-grinder (model TE-631/4, Piracicaba, SP, Brazil),
immersed in distilled water (20% w/v), and stirred at 25 ◦ C until the
gum was solubilized. The solution was filtered using a nylon membrane
(WHA7402002, Sigma-Aldrich) and precipitated with cold 95% ethanol
with a ratio of 1:3 (v/v). The precipitated CGP was separated by
centrifugation at 4000 x g, 4 ◦ C, for 10 min (Centrifuge model 5804R,
2
Food Bioscience 37 (2020) 100722
Eppendorf, Hamburg, Germany) and washed twice with ethanol to
remove the remaining impurities. The purified CGP was lyophilized
(Liotop model L101, Liobrás, São Carlos, SP, Brazil) and milled using the
mill-grinder, and the resulting powder was stored at room temperature
(25 ± 2 ◦ C) in airtight vials for a maximum of 6 months.
2.3. Preparation of edible coatings
Edible hydrogels were prepared by mixing 3 g of PVA (product
number 363138, Sigma Aldrich) and 1 g of CGP with 100 mL of distilled
water at 60 ◦ C with stirring. After solubilization, the mixture was cooled
to 25 ◦ C and 5 mL of 0.1 M H2O2 was added as an oxidizing agent, 10 mL
of 1 M H3PO4 as a catalyst and 0.1 g of mannitol (98% purity, Sigma
Aldrich) as a plasticizer were added and left at 25 ◦ C to slowly react for 4
days while the solvent was completed evaporated. The resulting matrix
was resolubilized in 50 mL of distilled water and homogenized (3000
rpm, 10 min) using an UltraTurrax® (model T25, Ika®-Werke GmbH &
Co., KG, Staufen, Germany) to obtain a CGP/PVA solution to produce
the edible coatings. To ensure the complete removal of the H2O2, the
mixture was incubated at 50 ◦ C for 1 h, with stirring. The presence of
H2O2 in the film-forming solution was determined using the method of
Noble and Gibson (1970). Samples of 100 μL of film-forming solution
were added to 1 mL of distilled water and the absorbance was deter­
mined at 240 nm using a UV–Vis spectrophotometer (BEL SPECTRO
model S-2000, BEL Engineering, Piracicaba, SP, Brazil). The amount of
H2O2 in the solution was calculated comparing the absorbance of the
sample with a calibration curve of H2O2 in distilled water (r2 = 0.995).
2.4. Characterization of the CGP/PVA edible coating
2.4.1. Fourier transform infrared spectroscopy (FTIR)
FTIR spectra were carried out on a FTIR spectrometer (Bruker Vertex
70, Bruker Co., Billerica, MA, USA) to characterize the presence of
specific chemical groups in the materials. Polymers (10 mg) and CGP/
PVA solutions (50 μL) were mixed thoroughly with potassium bromide
(100 mg) and compressed into pellets by applying pressure of 200 kg
cm− 2 in a hydraulic press (model 15T, Specac Inc., London, UK). FTIR
was done in attenuated total reflection mode between 400 and 4000
cm− 1 with 64 scans at a resolution of 4 cm− 1 and scan speed of 0.2 cm
s− 1 .
2.4.2. Scanning electron microscopy (SEM)
SEM was done at the Laboratório Multiusuário de Microscopia de
Alta Resolução at the Universidade Federal de Goiás (Goiânia, GO,
Brazil). For microstructural analysis, 5 mL of CGP/PVA solution was left
to dry at room temperature to obtain the edible coating which was cut
with a scalpel into strips of ~1 cm2 and fixed onto copper stubs. The
samples were gold-coated (Coating system model DESK V, Denton
Vacuum, Moorestown, NJ, USA), and observed using an accelerating
voltage of 15 kV (JEOL JSM-6610, Jeol Ltd., Welwyn Garden City,
Hertfordshire, England). Uncoated fruits (control) and fruit coated with
edible CPV/PVA coatings were cut with a scalpel in slices of 4 cm2 and
analyzed after 1 and 5 days of storage in a biosafety cabinet at 25 ◦ C and
30% relative humidity (RH), using the same conditions described above.
2.4.3. Viscosity analysis
Viscosity was measured as described by Saberi, Vuong, et al. (2016),
using oscillatory measurements in an analogue rotational viscometer
(model EEQ-9031, range 1–20,000 cP, EDUTEC, Curitiba, PR, Brazil).
The viscosity was determined at 60 rpm and room temperature using the
nº 2 spindle. Viscosity was measured in triplicate and the results were
expressed in mPa.s.
2.4.4. Light transmission of the CGP/PVA edible coating
Light transmission was measured according to the procedure re­
ported by Cao et al. (2018), with minor modifications. The light
B.R. Moreira et al.
transmittance of CGP/PVA edible coating was assessed using an ultra­
violet–visible (UV–Vis) spectrophotometer in the range of 200 to 800
nm, with 6 scans at a resolution of 10 nm and a scan speed of 250 nm
min− 1. The samples were fixed to a cuvette with a 1 cm slit width.
Samples were cut with a scalpel into strips 4x2 cm and tested in tripli­
cate. The thickness of the CGP/PVA edible films were determined using
a digital micrometer (model MDC-SX/25 mm, Mitutoyo Corp., Tokyo,
Japan) with minimal resolution of 0.001 mm and each strip was measure
10 times in different places. It was used with samples showing two
different values of thickness, 32 and 88 μm (±1 μm).
2.4.5. Water vapor permeability (WVP) and CO2 permeability
The WVP of the CGP/PVA edible coating was measured using the
gravimetric method described by Pérez-Gago and Krochta (2001).
CGP/PVA edible coating were cut with a scalpel into discs with ~1.5 cm
radius (area ≈6.6 cm2) and 80 ± 2 μm thickness were sealed using an
open metal cap on the mouth of weighing bottles containing 25 g of
anhydrous CaCl2. These weighing bottles were conditioned in a desic­
cator with saturated NaCl solution to achieve 75% RH. Weight changes
were measured for 6 days at 12 h intervals. The slopes of the weight gain
(Δm) versus time (Δt) were obtained. Using these values, the water vapor
transmission rate (WVTR) and WVP were calculated using the following
equations:
△m
WVTR = (1)
(△t • A)
( )
X
WVP g • s− 1 • m− 1 • Pa− 1 = WVTR • (2)
△p
where A (m2) indicates the exposed sample area (0.00066 m2), X (m)
indicates the average thickness of the CGP/PVA edible coating (0.00008
m) and Δp (Pa) indicates the difference in water vapor pressure between
the two sides of the sample (1753.55 Pa).
For the evaluation of CO2 permeability, films with thickness of 281
± 2 μm were cut with a scalpel into discs of ~38 cm2. Prior to the
determination, samples were conditioned at 25 ◦ C for 48 h in a desic­
cator with saturated K2CO3 solution to achieve 50% RH. The CO2 (99%
purity, White Martins Gases Industriais Ltda., Rio de Janeiro, RJ, Brazil)
permeability was measured with the Perme Vac-V1 gas permeability
measuring device (model PermeVac-V1, Medford, MA, USA), running
with a time interval of 10 s and a vacuum for 3 h. Testing was based on
the differential pressure method to determine the gas transmission rate
of films using the ASTM D1434-82 standard (ASTM, 2009). Each sample
was measured in triplicate.
2.4.6. Mechanical properties
The mechanical properties of the CGP/PVA edible coatings (thick­
ness of 200 ± 0.3 μm), including tensile strength (TS) and elongation at
break (EB), were determined in a texture analyzer (model TA1, Lloyd
Instruments/Ametek, Largo, FL, USA) with a 50 N load cell equipped
with tensile grips (model A/TG). Prior to the determination, samples
were cut with a scalpel into rectangular slices of 15x90 mm and
conditioned at 25 ◦ C for 24 h in a desiccator containing a saturated
K2CO3 solution to achieve 50% RH, in accordance with ASTM D882-02
(2002). Tests were done using 10 replicates and the initial grip separa­
tion and cross-head speed were set at 50 mm and 300 mm min− 1,
respectively. TS and percentage of EB were evaluated. Each sample used
was previously inspected and those containing any defect such as air
bubbles, holes, and tears were rejected.
2.5. Bioactivation of CGP/PVA edible TCWDE from Trichoderma
asperellum T00
The production of TCWDE was done as described by Fernandes et al.
(2013). Briefly, T. asperellum (obtained from the Laboratory of
3
Food Bioscience 37 (2020) 100722
Enzymology’s culture collection at the Federal University of Goias) was
inoculated in Trichoderma liquid enzyme production medium (0.1% of
peptone; 0.03% of urea; 0.2% of KH2PO4; 0.14% of (NH4)2SO4; 0.03% of
MgSO4; 0.03% of CaCl2; 1 mL of 0.01% trace elements solution),
enriched with 0.02% (w/v) glucose and 0.5% (w/v) chitin from shrimp
shells (C7170, Sigma-Aldrich). The culture was incubated at 28 ◦ C for 6
days and harvested by filtration using Whatman filter paper
(WHA1001325, Sigma Aldrich). The filtrate containing the TCWDE was
dialyzed (cellulose membranes with nominal molecular weight cut-off
14 kDa, D9277, Sigma Aldrich) against water and lyophilized. The ac­
tivity of the enzymes produced by T. asperellum was evaluated according
to methodology described by Silva et al. (2011). The chitinase activity
was determined using colloidal chitin as the substrate. An aliquot of 500
μL of TCWDE solution (20 mg mL− 1) was mixed with 500 μL of colloidal
chitin and incubated for 2 h at 37 ◦ C. The reaction mixture (250 μL) was
added to 50 μL of 0.3 mol L− 1 K2B4O7 and incubated for 3 min at 100 ◦ C.
Then, 1.5 mL of 0.1% (w/v) 4-(dimethylamino) benzaldehyde solution
(DMAB, D2004, Sigma-Aldrich) was added and the samples were incu­
bated for 10 min at 37 ◦ C. The absorbance was measured at 587 nm and
one unit of enzyme (U) was defined as the amount of enzyme necessary
to produce 1 μmol of reducing sugar in 1 h at 37 ◦ C.
N-acetylglucosaminidase activity was done by monitoring the rate of
formation of p-nitrophenol from 4-nitrophenyl N-acetyl-β-D-glucosami­
nide (pNglcNAc, N9376, Sigma Aldrich). An aliquot of 50 μL of TCWDE
solution (20 mg mL− 1) was mixed with 100 μL of 5 mmol L− 1 pNglcNAc
solution and 350 μL of sodium acetate buffer (50 mmol L− 1, pH 5.2). The
mixture was incubated for 15 min at 37 ◦ C. Sodium carbonate solution
(1 mL of 0.5 mol L− 1) was added and the content of p-nitrophenol
released was measured at 450 nm. One unit of enzyme (U) was defined
as the amount of enzyme that releases 1 μmol of p-nitrophenol in 1 min
at 37 ◦ C.
The activity of β-1,3-glucanase was evaluated using laminarin
(L9634, Sigma Aldrich) as the substrate. Samples of 50 μL of TCWDE
solution (20 mg mL− 1) were mixed with 100 μL of 0.25% (w/v) lami­
narin solution in acetate buffer (50 mmol L− 1, pH 5.2). The mixture was
incubated for 1 h at 40 ◦ C and the amount of reducing sugars formed was
determined at 550 nm (Miller, 1959). One unit of enzyme (U) was
defined as the amount of enzyme that produces 1 μmol of reducing sugar
in 1 min at 40 ◦ C.
CGP/PVA edible coatings were bioactivated by mixing 10 mg of
TCWDE with 1 mL of CGP/PVA solution. The system was stirred for 10
min at 25 ◦ C and then used as a coating for fresh strawberries (Fragaria
ananassa). The activity of the CGP/PVA/TCWDE edible coatings were
evaluated using the method of Silva et al. (2011), using 50 μL of
CGP/PVA/TCWDE solution. The CGP/PVA/TCWDE film-forming solu­
tion had the following enzyme activities: (1) 0.6 U mL− 1 for chitinase,
(2) 144 U mL− 1 for N-acetylglucosaminidase and (3) 6 U mL− 1 for β-1,
3-glucanase.
2.6. Biological tests
To determine the efficiency of CGP/PVA and CGP/PVA/TCWDE
edible coatings with respect to retarding or inhibiting fungal growth,
tests were done using these materials as edible coatings for fresh
strawberries. Strawberries (average weight: 14 ± 0.5 g) were purchased
in a local market and immediately used in the biological tests. Prior to
the beginning of the experiments, damaged, nonuniform, unripe, or
overripe fruits were discarded.
2.6.1. Coating and storage conditions of the strawberries
The coating of strawberries (including the sepal and pedicel) was
done as described by Chlebowska-Smigiel et al. (2008). The fruits were
washed with running water and rinsed with distilled water, drained and
air dried on a stainless-steel screen for 30 min prior to applying the
coating. To evaluate the antifungal effect of the CGP/PVA edible
coating, the CGP/PVA or CGP/PVA/TCWDE edible hydrogels were
B.R. Moreira et al.
spread over the entire strawberries surface using a sterile brush. The
covered fruits were allowed to dry in a biosafety cabinet for 2 h (25 ◦ C,
36% RH) to ensure polymerization of the edible coating.
A solution of Penicillium sp. (obtained from the Laboratory of Enzy­
mology’s culture collection at the Federal University of Goias) was
prepared in saline buffer and the numbers of spores were determined
using a Neubauer chamber (Hecht Assistent®, Glaswarenfabrik Karl
Hecht GmbH & Co. KG, Sondheim vor der Rhön, Germany). For the tests,
an aliquot of 200 μL of Penicillium spore solution (107 spores mL− 1) was
spread over the surface of each strawberry (excluding the sepal and
pedicel structures) using a brush and allowed to dry in the biosafety
cabinet for 1 h. The fruits were stored in the biosafety cabinet (25 ◦ C,
36% RH) for 5 days. A total of 15 strawberries were inoculated/exper­
iment. For the biological tests, strawberries were organized in three
groups:
Group A: CGP/PVA coated group
Group B: CGP/PVA/TCWDE coated group
Group C: control (uncoated strawberries)
2.6.2. Structural appearance and weight loss
A macroscopic evaluation was carried out to determine the fruit
appearance and fungal incidence during 5 days of storage. All groups
were stored at room temperature and the images were captured with a
fixed (10 cm of distance from subject) photographic camera (model
GU46 6 GG, Samsung, London, UK), using indoor natural lighting con­
ditions. In addition, after 5 day of storage samples from coated and
uncoated strawberries were analyzed using SEM as described in section
2.3.2.
The weight loss was measured as described by Duan et al. (2011).
The percentage of weight loss (WL) was calculated as coated weight
(FW) of the fruit at each sampling time divided by the initial coated
weight (IW) of the fruit (after coating and drying), using the following
equation:
FW
WL(%) = • 100 (3)
IW
2.7. Statistical analysis
All measurements were done at least in triplicate. Results were
expressed as mean ± standard deviation (X ± SD). Variance analysis
(one-way ANOVA) and Tukey’s test were used to define differences in
mean values. The variance analyses were carried out using Statistica 6.0
software (StatSoft Inc., Tulsa, OK, USA). The means of treatments with
the same letters were not significantly different (p ≤ 0.05).
Fig. 1. Macroscopic appearance (a) and microstructural surface (b) of the CGP/PVA edible coating.
4
Food Bioscience 37 (2020) 100722
3. Results and discussion
3.1. FTIR spectroscopy
The CGP/PVA edible coating was visually analyzed and no fractures
or ruptures were observed after drying. The film was visually homoge­
neous and did not show opaque or different colored zones or the pres­
ence of insoluble particles (Fig. 1a). SEM images showed that the surface
of the CGP/PVA edible coating had a homogeneous and regular struc­
ture, without aggregates or the presence of pores (Fig. 1b).
The FTIR spectra of the CGP, PVA and the CGP/PVA edible coating
are shown in Fig. 2. The FTIR spectrum of CGP showed the major peaks
associated with this polysaccharide. There is a strong broad band around
3435 cm− 1 assigned to the stretching vibration of hydroxyl groups and a
band at 2934 cm− 1 assigned to the C–H broad alkyl stretching band
(Fig. 2a). The band interval between 1100-700 is commonly referred as
“fingerprint region” related to the polysaccharide structure. The peaks at
~1080 and 710 cm− 1 were due to the stretching vibrations of –C–O–C
from glycosidic bonds and stretching vibrations of –OH bending
resulting from the pyranosidic structures of CGP (Pavia et al., 2010;
Pitombeira et al., 2015). A peak around 1640 cm− 1 is associated with the
asymmetric deformation of the –OH water groups (Lima et al., 2018).
FTIR spectrum of pure PVA (Fig. 2b) showed a C–H broad alkyl
stretching band around 2900-3000 cm− 1 and typical strong hydroxyl
bands for free alcohol (nonbonded –OH stretching band at 3600-3650
cm− 1), and a hydrogen bonded band (around 3200-3570 cm− 1) (Hassan
& Peppas, 2000; Peppas, 1977; Reis et al., 2006). An important ab­
sorption peak was found at a frequency of 1096 cm− 1. This vibrational
band is mostly attributed to the crystallinity of the PVA related to a
carboxyl stretching band (C–O, around 1090-1150 cm− 1) (Coates, 2000;
Mansur et al., 2002). This absorption band has been used as an
Fig. 2. FTIR spectra of (a) CGP, (b) PVA and (c) CGP/PVA edible coating.
B.R. Moreira et al.
assessment tool for the PVA structure because it is a semi-crystalline
synthetic polymer capable of forming some domains depending on
several process parameters. In addition, the vibrational frequency at
1736 cm− 1 of pure PVA was assigned to the C– – O stretching of the ac­
etate groups in the PVA structure (Malathi et al., 2010).
The spectrum shown in Fig. 2c confirmed the formation of CGP/PVA
blends. The disappearance of the hydroxyl groups (1600-1650 cm− 1)
and the presence of bands of the acetal ring (C–O–C) and the ether (C–O)
linkages (1000-1130 cm− 1) were probably due to the covalent bond
formed between oxidized PVA and CGP (Pavia et al., 2010; Vlachos
et al., 2006).
3.2. Viscosity behavior
The viscosity behavior of coating-forming solutions is an important
factor that can affect the spread-stability, thickness, and uniformity of
the liquid coating layer, in addition to directly interfering with me­
chanical properties, processing design, and applications (Ma et al.,
2017). The CGP/PVA edible hydrogel had a viscosity of 111 ± 1 mPa s.
This value was consistent with those reported for films based on gummy
polysaccharides (Simas-Tosin et al., 2010). Nevertheless, there is a range
of viscosity values for edible coatings, which are dependent on the na­
ture and structure of the polysaccharides used in the hydrogel formu­
lations. Intermolecular interactions will occur with a high frequency in
linear polysaccharides, which will allow water molecules to move freely
in the solution, thus decreasing the apparent viscosity. On the other
hand, interactions with water molecules will predominate with
branched polysaccharides and cause a reduction in the amount of free
water in the solution, which increases the apparent viscosity. The hy­
drophilicity of the sugar components will also influence the viscosity of
the resultant hydrogel.
Despite the different viscosity behaviors of polysaccharide-based
edible coatings, several researchers have shown that when the viscos­
ity of the coating-forming solution is too high, there will be disconti­
nuities in the final films (Cao et al., 2018; Ma et al., 2017; Saberi, Vuong,
et al., 2016). Sothornvit and Krochta (2005) reported that plasticizers
can restrict interactions of the polymer chain, creating a less organized
film structure, which may decrease the viscosity and also interfere with
the film structure. Mannitol (0.1% w/v) was used as a plasticizer agent.
As the amount of plasticizer added to the formulation was low, no
interference in the structure of the CGP/PVA edible coating was
observed, indicating that polysaccharide branching and the interactions
between the polymer chains are flexible enough to maintain a stable
network.
3.3. Light transmission
The transparency of edible coatings is an important property for
determining the applications of a material, mainly when it will be
applied as a coating for food surfaces or for amending the appearance of
the final product (Maran et al., 2013). In general, it is desirable that film
packaging and edible coatings show high transparency and lightness.
The regular light transmission spectra of the CGP/PVA edible coat­
ings with two different thicknesses (32 and 88 μm) are shown in Fig. 3.
The film thickness affected the light transmission statistically signifi­
cantly in the ultraviolet range (200–400 nm), with a reduction of about
25% in light transmission for the thicker film (88 μm). However, in the
visible spectrum (400–700 nm) no statistically significant differences in
light transmission were observed in the CGP/PVA edible coatings
despite the 2.8 fold increase in the thickness. It is possible that the
structural network of the CGP/PVA edible coating is regularly orga­
nized, which allows light transmission independent of the thickness of
the material. The CGP/PVA edible coatings were transparent with a
transmittance of 88% in the visible range and showed excellent optical
characteristics as can be seen in Fig. 1a in which the writing covered by
the CGP/PVA edible coating (88 μm) is visible.
5
Food Bioscience 37 (2020) 100722
Fig. 3. Light transmission curve of the CGP/PVA edible coating with different
thicknesses.
The light transmission profile was consistent with those reported for
polysaccharide-based films and coatings. Silva et al. (2016) reported
light transmission values around 70% for CGP/PVA films with thick­
nesses between 140 and 200 μm. Ortega-Toro et al. (2017) found light
transmission values between 69 and 84% for antifungal starch-based
edible films containing Aloe vera. Saberi, Vuong, et al. (2016) evalu­
ated biodegradable edible films based on pea starch and guar gum and
obtained transmission values of 78 to 90%. This profile of light trans­
mission for the CGP/PVA edible coating allows a multitude of biotech­
nological applications for this material, since this coating does not
compromise the visualization of the product, an important parameter for
successful commercialization.
3.4. WVP and CO2 permeability
The WVP of coating materials is one of the most important properties
of edible coating and films for food packaging applications, mainly
because of the importance of the water in deteriorative reactions that
directly influence the shelf-life of food products (Cerqueira et al., 2009;
Spotti et al., 2016). Therefore, the lower the WVP value the better the
barrier property (Du et al., 2016). The CGP/PVA edible coating showed
a WVP of 7 x 10− 11 g m− 1s− 1Pa− 1 which was lower than those values
shown for other polysaccharide-based films. Zhang et al. (2016) re­
ported WVP values of 17 x 10− 11 g m− 1s− 1Pa− 1 for edible films based on
gum ghatti. In another study, corn starch-based films containing glycerol
had a WVP value of 8.7 x 10− 11 g m− 1s− 1Pa− 1 (Garcia et al., 2006).
Bravin et al. (2006) reported values of 20.5 x 10− 11 g m− 1s− 1Pa− 1 for
WVP of starch and methylcellulose films. These differences in the WVP
values might result from the diversity in chemical composition of
coating solutions or crystallinity of the films.
Polysaccharide edible coatings have a hydrophilic nature, which
provides good barriers to oxygen, carbon dioxide, and lipids (Biliaderis
et al., 1999; Cerqueira et al., 2009). Carbon dioxide is important for
living tissue respiration and a higher value of CO2 permeability can
delay fruit softening. Therefore, a low rate of CO2 permeability in edible
coatings is desirable. Edible CGP/PVA coatings showed a CO2 perme­
ability of 363 cm3 m− 2 day− 1 at 0.1 Mpa− 1, a value lower than those
reported by Carvalho et al. (2017) for cellulose acetate films (5600 cm3
m− 2 day− 1 at 0.1 Mpa− 1). These results confirmed the network cohesion
of CGP/PVA edible coating, which can act to modify the internal at­
mosphere in the fresh-cut fruits and vegetables and cause retardation of
the biochemical reactions linked to deterioration (Tzoumaki et al.,
2009).
B.R. Moreira et al. Food Bioscience 37 (2020) 100722

3.5. Mechanical properties

The mechanical properties of CGP/PVA edible coatings were

assessed by measuring their tensile strength and elongation upon
breaking. Tensile strength (TS) represents the maximum tensile stress
before breaking. Elongation upon breaking (EB) is a measure of film
stretching capacity. The CGP/PVA edible coating showed values of 0.04
MPa for TS and 147% for EB. After comparing the mechanical properties
of the CGP/PVA edible coating with other films, it was observed that the
resulting TS value was lower than those obtained for other poly­
saccharides while the EB values were significantly higher (Cazón et al.,
2017; Cerqueira et al., 2009; Chiumarelli & Hubinger, 2012; Davidovic
et al., 2018; Souza et al., 2012; Spotti et al., 2016). According to Seyedi
et al. (2014), coatings and films with low TS and high EB tend to be less
fragile, especially when plasticizers are used in the formulations. In the
case of CGP/PVA edible coatings, the mechanical results can be related
to the mannitol penetration into the polymer matrix and reduction of
inter-chain interactions which led to a high flexibility, showing that it
underwent fracture with a slow and sustained pace. The
three-dimensional organization of PVA and CGP also provided high
flexibility characteristics to the final edible coating.

3.6. Biological tests

Edible coatings are intended to extend the shelf-life by acting as

semi-permeable barriers, reducing moisture and solutes losses, gas ex­
change, respiration and oxidative reaction rates, as well as suppressing
physiological disorders on fresh-cut fruits (Gol et al., 2013; Hassan et al.,
2018). It was also reported that edible coatings may act as carriers for
food additives such as antimicrobial agents, on food surfaces, to confer
bioactivity against microorganisms (Davidovic et al., 2018; Ribeiro
et al., 2007). Therefore, aiming to determine the efficiency of CGP/PVA
edible coating in extending the shelf-life of fresh strawberries, storage
tests were done using uncoated (control) and coated fruits (groups A and
B).
In the macroscopic evaluation, it was observed that the strawberries
coated with CGP/PVA edible coatings (groups A and B) showed a delay
in the deterioration process compared to those uncoated (control). The
coated fruits were visually brighter and more reddish compared to the
controls, while the uncoated strawberries showed a visible weight loss,
becoming withered and smaller (Fig. 4).
According to Saberi, Thakur, et al. (2016) edible films provide
reinforcement of natural layers to prevent weight losses, allowing a
controlled interchange of important gases involved in the respiration of
plant tissues, such as carbon dioxide, ethylene and oxygen. Weight loss
is an important indicator of fruit freshness loss. The assessment of
weight loss showed that the behavior of the control group strawberries
was significantly different from the coated ones (p < 0.05). Group C
showed a weight loss of 59% (±0.3), indicating that the uncoated
strawberries lost 31% more weight than group A. In addition, the
presence of immobilized enzymes did not interfere with the weight loss
of the coated fruits with a 41% (±4) weight loss for those strawberries
from group A and 45% (±1) for strawberries from group B, after 5 days
of storage.
According to Duan et al. (2011), migration of water from the fruit to
the environment is the main cause of weight loss during storage. The use
of edible coatings can reduce the weight loss because they act as an extra
layer that also coats the stomata in the fruit, leading to a decrease in
transpiration and respiration rates (Guerreiro et al., 2015; Ortega-Toro
et al., 2017).
The decrease in weight loss in the strawberries from groups A and B
is probably related to coating composition. The entrapment of moisture
between the CGP/PVA coating and fruit surface might reduce the
dehydration rate of the fruits, thus increasing the strawberries shelf-
lives. Despite the entrapped moisture being a factor that can favor the
growth of fungus that were already on the fruit before the coating
Fig. 4. Macroscopic appearance of strawberries on the first and fifth storage
days. (a) uncoated strawberries (group C); (b) strawberries coated with CGP/
PVA coating (group A); and (c) strawberries coated with the edible CGP/PVA/
TCWDE edible coating (group B).
process, the natural antifungal activity (Cruz et al., 2017) shown by CGP
probably inhibited the possibility of fungal development in the coated
fruits, even without the presence of TCWDE in the formulation of the
edible coating (Fig. 6).
SEM images showed the surfaces of uncoated and coated straw­
berries on the first and fifth storage days at room temperature (Fig. 5).
The structure of uncoated strawberries (Fig. 5a) underwent changes over
time, showing a transition from a smooth to a rough surface as a
consequence of the water loss. On the other hand, the structure of the
strawberries that received the edible coating remained unchanged
(Fig. 5b), showing that the CGP/PVA coating delayed the water-loss
related deterioration process. These results were consistent with those
reported in the literature (Gol et al., 2013; Li et al., 2017; Maqbool et al.,
2011), confirming that coatings on fruits and vegetables can serve as a
semipermeable barrier and cause a reduction in water loss and oxidation
reactions, which delay the product’s senescence process.
The efficiency of the antifungal growth effects of the CGP/PVA edible
coating containing TWCDE were evaluated in tests using Penicillium sp.,
a common fungus pathogen in fruits, as a contaminant. Strawberries
were examined over 5 days of storage (Fig. 6). The visible microbial
growth on the fruit was characterized as brown spots and softening in
the injured zone.
Coated strawberries (group A and B) maintained a uniform appear­
ance, with minor loss of mass and color variation. They did not show
signs of visible fungal decomposition during the storage period (Fig. 6b
and c). However, the uncoated strawberries were completely vulnerable

6
B.R. Moreira et al. Food Bioscience 37 (2020) 100722

Fig. 5. Micrographs of the strawberries surface on the first and fifth storage days: (a) uncoated strawberries; (b) strawberries coated with the edible CGP/
PVA coating.

to the deteriorative actions of the inoculated fungus and showed weight

loss, size reduction, and color alteration in addition to signs of fungal
contamination and advanced deterioration (Fig. 6a).
The CGP/PVA edible coating without TCWDE enzymes (Fig. 6b) was
apparently enough to avoid fungal development on the strawberries.
These results could be explained as a function of the physical barrier
provided by the coating layer and also due to the intrinsic antimicrobial
activity of CGP as reported by Cruz et al. (2017), who observed that CGP
powder caused cell wall changes in Aspergillus fumigatus and Penicillium
spp.

4. Conclusions

The blend of CGP and PVA using hydrogen peroxide as an oxidizing

agent can produce a biodegradable hydrogel that could be used to pre­
pare edible coatings with good mechanical properties and barrier fea­
tures. CGP/PVA hydrogels showed a viscosity of 111 mPa.s, which was
enough to form an edible and water-soluble coating with a homoge­
neous microstructural surface without the presence of aggregates or
pores. The CGP/PVA edible coating showed 88% light transmission, thus
allowing diverse applications. In addition, the biological tests showed
that a CGP/PVA edible coating is efficient at preventing weight loss and
fungal deterioration of strawberries. The weight loss process of the
strawberries coated with the CGP/PVA edible coating was delayed. SEM
showed that the structure of the coated strawberries was preserved after
5 days of storage at room temperature. Fungal growth inhibition tests
showed that CGP/PVA edible coatings were efficient as an antimicrobial
barrier even without bioactivation using TCWDE, preventing fungal
contamination, and consequently increasing the shelf-life of fresh
strawberries. Therefore, the combination of all of these characteristics
allows for the use of CGP/PVA edible coatings as an ecofriendly and
bioactive food packaging material that contributes to avoiding fungal
contamination and water loss of fresh fruits during storage.

Author contributions
Fig. 6. Fungal growth inhibition of the CGP/PVA edible coatings in straw­
berries inoculated with Penicillium spp. (a) uncoated strawberries (group C); (b) Bruna Moreira: methodology, investigation, validation, formal
strawberries coated with edible CGP/PVA coating (group A); and (c) straw­ analysis, writing - original draft.
berries coated with edible CGP/PVA/TCWDE coatings (group B).

7
B.R. Moreira et al.
Marcos Pereira-Junior: methodology, investigation, validation,
formal analysis, writing - original draft.
Kátia Fernandes: Conceptualization, writing - review & editing, su­
pervision, funding acquisition.
Karla Batista: Conceptualization, formal analysis, writing - review &
editing, visualization, supervision, project administration, funding
acquisition.
Declaration of competing interest
The authors confirm that they have no conflicts of interest with
respect to the work described in this manuscript.
Acknowledgements
This work was supported by the Conselho Nacional de Desenvolvi­
mento Científico e Tecnológico – Brazil (CNPq) [Process number
402468/2013-9]. M.A. Pereira-Júnior and B.R. Moreira would like to
thank the Coordenação de Aperfeiçoamento de Pessoal de Nível Supe­
rior – Brazil (CAPES) for fellowship support [Finance code 001].
Appendix A. Supplementary data
Supplementary data to this article can be found online at https://doi.
org/10.1016/j.fbio.2020.100722.
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