Food Hydrocolloids 89 (2019) 682–690

Contents lists available at ScienceDirect

Food Hydrocolloids
journal homepage: www.elsevier.com/locate/foodhyd

Preparation and characterization of konjac glucomannan-based T

bionanocomposite film for active food packaging
Chunhua Wua,b, Yuanzhao Lia, Yu Dua, Lin Wanga, Cailing Tonga, Yaqin Hub, Jie Panga,∗∗,
Zhiming Yana,∗
a
College of Food Science, Fujian Agriculture and Forestry University, Fuzhou, 350002, China
b
College of Biosystems Engineering and Food Science, Zhejiang University, Hangzhou, 310058, China

A R T I C LE I N FO A B S T R A C T

Keywords: Novel active bionanocomposite films were prepared by incorporating chitosan(CS)/gallic acid (GA) nano-
Chitosan/gallic acid nanoparticles particles (CGNPs) into a konjac glucomannan (KGM) film. CGNPs with a GA loading efficiency of 78 ± 2.3%
Konjac glucomannan were obtained through ionotropic gelation method, and they presented a spherical morphology with a diameter
Bionanocomposite film range of 80–100 nm. The influences of the CGNPs content on the structural, morphological, mechanical, barrier,
Antimicrobial properties
thermal and antimicrobial properties of KGM/CGNPs films were discussed. The rheological results of film-
forming solutions revealed that the CGNPs interacted with KGM through hydrogen bonds in a bionanocomposite
matrix, which was coincidence with the fourier transform infrared spectroscopy, X-ray diffraction and thermal
analysis results. The microstructure of the films showed that the introduced 5–10% CGNPs appeared to be
homogeneously dispersed within the KGM film matrix, thereby reducing the free volume of the composite
matrix, and improving the final bionanocomposite films' mechanical, and barrier properties significantly
(p < 0.5). In comparison with a pure KGM film, KGM/CGNPs bionanocomposite films showed an excellent
antimicrobial activity against food-borne pathogens such as gram-positive (Staphylococcus aureus) and gram-
negative (Escherichia coli O157:H7) bacteria, because of antimicrobial effectiveness of the CGNPs. Therefore,
KGM/CGNPs bionanocomposite films show potential for applications in active food packaging materials.

1. Introduction
With the environmental concerns and food safety problems caused
by petrochemical-based plastic packaging materials, a tremendous
growing of the research interest has shifted to natural biopolymer-based
edible packaging materials (Dehghani, Hosseini, & Regenstein, 2018;
Liang, Sun, Cao, Li, & Wang, 2018). Nature has provided various nat-
ural biopolymers such as proteins (e.g. soy isolate protein, zein, casein,
and gelatin), polysaccharides (e.g. cellulose, chitosan, starch and glu-
comannan) and lipids (Cazón, Velazquez, Ramírez, & Vázquez, 2017;
Dehghani et al., 2018; Gómez et al., 2016). Amongst these biopolymers,
konjac glucomannan (KGM) has been considered as a good candidate
for edible packaging materials (Li et al., 2015; Wang et al., 2017). KGM
is a natural polysaccharide extracted from the tubers of Amorphophallus
konjac plant (Yuan et al., 2018). This polysaccharide has been widely
explored in the preparation of biodegradable or edible films because of
its non-toxicity, excellent film-forming ability and biodegradability
(Wang et al., 2017). However, the widespread application of pure KGM
films is limited by their relatively poor water resistance and low me-
chanical and antibacterial properties (Du, Yang, Ye, & Li, 2013).
To overcome these shortcomings of biopolymer films, a new class of
materials namely bionanocomposites (biopolymer matrices reinforced
with nanofillers) has been introduced as a promising option (Azeredo,
Rosa, & Mattoso, 2017; Rhim, Park, & Ha, 2013). The addition of ap-
propriate nanofillers may improved the water barrier properties, me-
chanical properties, thermal stability and other functional properties of
the nanocomposite films (HPS et al., 2016). Amongst these nanofillers,
chitosan nanoparticles (CSNPs) formulated through ionotropic gelation
are considered as a attractive reinforcement filler for edible films and/
or food packaging (Hosseini, Rezaei, Zandi, & Farahmandghavi, 2015;
Lorevice, Otoni, Moura, & Mattoso, 2016; Wu et al., 2016). The in-
corporation of CSNPs enhances the physicochemical properties of
polysaccharide-based films, such as hydroxypropyl methylcellulose
(Moura et al., 2009), starch (Chang, Jian, Yu, & Ma, 2010), tara gum
(Antoniou, Liu, Majeed, & Zhong, 2015), and pectin (Lorevice et al.,
2016). However, the antibacterial activity of pure CSNPs is also limited

∗
Corresponding author.
∗∗
Corresponding author.
E-mail addresses: pang3721941@163.com (J. Pang), 45765545@qq.com (Z. Yan).

https://doi.org/10.1016/j.foodhyd.2018.11.001
Received 12 June 2018; Received in revised form 1 November 2018; Accepted 1 November 2018
Available online 15 November 2018
0268-005X/ © 2018 Elsevier Ltd. All rights reserved.
C. Wu et al. Food Hydrocolloids 89 (2019) 682–690

(Antoniou et al., 2015). An emerging approach to solve this problem is handle. Film-forming solutions were then centrifuged at 4000 rpm for
by incorporating plant phenolic extracts that show potent antimicrobial 10 min to remove undissolved matter and air bubbles. Then, 25 mL of
activities in food systems and their intake can contribute to human the solution was poured into plastic Petri dishes and dried at 55 °C for
health. Gallic acid (GA), a well-known natural phenolic acid with strong 24 h. The obtained films were conditioned at 53 ± 1% relative hu-
antimicrobial and antioxidant activities, has been successfully in- midity and 25 ± 1 °C for at least 3 days in a controlled environment
corporated into biodegradable films to fabricate active packaging or chamber.
edible films (Hager, Vallons, & Arendt, 2012; Sun, Wang, Kadouh, &
Zhou, 2014). However, up to now, limited information about the use of
2.4. Characterization of CGNPs and bionanocomposite films
CSNPs as GA carriers (CGNPs) has been available (Lamarra, Rivero, &
Pinotti, 2016). To the best of our knowledge, the incorporation of
The morphological characteristics of CGNPs were examined by a
CGNPs as reinforcing agents in KGM-based bionanocomposite films has
high-performance digital imaging transmission electron microscope
yet to be reported.
(TEM, JEOL 2100, Hitachi High-Technologies Corp., Tokyo, Japan).
This study aimed to develop KGM/CGNPs bionanocomposite films
The particle size, polydispersity index (PDI) and Z-potential of the na-
and evaluate the effects of CGNPs concentration on the microstructural,
noparticles were measured with a Zetasizer Nano ZS90 (Malvern
physicochemical and antibacterial aspects of the produced films. Our
Instruments, UK). The encapsulation efficiency (EE) of CGNPs were
findings suggested that KGM/CGNPs bionanocomposite films could be
determined according to the method reported by Lamarra et al. (2016).
considered as food packaging materials.
The rheological properties of the film-forming solutions were de-
termined using an Anton Paar MCR 301 rheometer (Anton Paar
2. Materials and methods
Instruments Inc., Austria) equipped with a parallel-plate geometry
(diameter = 50 mm) at 25 °C and at a shear rate of 0.1–100 s−1.
2.1. Materials
Dynamical frequency sweeps were carried out by applying 1% strain
within the linear viscoelastic range over a frequency range of
Konjac glucomannan (KGM) (purity of 95%, viscosity: 1% solution,
0.01–100 Hz. The storage or elastic modulus (G′, Pa) and the loss or
30 °C, ≥35,000 MPa s) was supplied by San Ai Konjac Food Co. Ltd.
viscous modulus (G″, Pa) were calculated as a function of dynamic
Chitosan (CS) (> 75% degree of deacetylation), GA, and sodium tri-
frequency.
polyphosphate (TPP) were purchased from Sigma-Aldrich Co. Ltd. (St.
Fourier transform infrared spectroscopy (FT-IR) spectra were ob-
Louis, MO, USA). Other analytical grade chemical reagents were pur-
tained using a Thermo Nicolette 6700 spectrophotometer (Thermo
chased from Sinopharm Group Chemical Reagent Co. Ltd. (China). Food
Fisher Scientific Co., Ltd., MA, USA) from 400 cm−1 to 4000 cm−1 at a
borne pathogenic microorganisms, including the gram-negative bac-
resolution of 4 cm−1. The X-ray diffraction (XRD) patterns of the films
teria, Escherichia coli O157:H7 (ATCC25922), and the gram-positive
were obtained by a Bruker AXS D8 Advance X-ray diffractometer
bacteria, Staphylococcus aureus (ATCC 25923-3), were provided by
(Bruker Inc., Germany) equipped with Ni-filtered Cu Kα radiation.
Hope Bio-Technology Co. Ltd. (Qingdao, Shandong, China).
Thermal analysis (TA) was conducted by using a TA instruments (TGA
Q500, Newcastle, DE, USA) at a rate of 10 °C/min in a nitrogen atmo-
2.2. Preparation of GA-loaded CS nanoparticles (CGNPs)
sphere. The surface and cross-section microstructures of films were
obtained using a scanning electron microscope (SEM, Japan Electron
GA-loaded CGNPs were prepared through ionic gelation in ac-
Optics Laboratory Co., Ltd, Tokyo, Japan). Film samples were mounted
cordance with previously described methods (Lamarra et al., 2016; Wu
on aluminum stubs using a double-sided adhesive carbon tape and
et al., 2016) with some modifications. In brief, CS was dissolved in 1%
sputtered with a thin layer of gold. The topography of the films was
aqueous acetic acid to form a polymer concentration of 5 mg/mL. The
analysed by a digital Instrument atomic force microscope (AFM, Bruker
corresponding amounts of GA were added to the CS solutions until the
Daltonics Inc., USA). All images were acquired in tapping mode at a
final concentration of 30 mgGA/gCS was reached. The resulting mixture
typical scan rate of 1 Hz with a scan range of 1 μm. The roughness
was stirred for 2 h at 25 °C. TPP was obtained from an aqueous solution
parameters such as average roughness (Ra) and root mean square
at a concentration of 1 mg/mL. Afterwards, the TPP solution was added
roughness (Rq) were obtained by the NanoScope software (Bruker/
dropwise to the CS/GA solution under vigorous magnetic stirring
Veeco, Inc., Santa Barbara, CA, USA). The transmittance spectra of the
(500 rpm) for 3 h at 25 °C. CGNPs were collected through centrifugation
films were obtained by a UV-2600 spectrophotometer (Shimadzu
at 8000 rpm for 1 h at 4 °C and subsequently washed several times with
Scientific Instruments, Inc., Kyoto, Japan) equipped with a quartz
deionised water. CGNPs were re-suspended in water, sonicated and
window plate. The spectra were recorded at room temperature in a step
lyophilised for further use. The encapsulation efficiency (EE) of GA-
of 0.5 nm in the range of 200–600 nm.
loaded CSNPs was determined by using a UV-2600 spectrophotometer
The tensile strength (TS, MPa) and elongation to break (EB,%) of the
(Shimadzu Scientific Instruments, Inc., Kyoto, Japan) in accordance
films were determined with a PARAM XLW (M) auto tensile tester
with previously described methods (Lamarra et al., 2016). In our study,
(Jinan Labthink Technology Company, China) in accordance with
EE was 78.0 ± 2.3%.
American Society for Testing and Materials (ASTM) standard methods
(D882-09, 2009). The water vapour permeability (WVP) of the films
2.3. Preparation of bionanocomposite films
was determined gravimetrically at 25 ± 1 °C according to ASTM E96
(E96/E96M-05 (2005),) standards. The antibacterial activity of the
The bionanocomposite films were prepared by solution casting
films was determined by the inhibition zone method in accordance with
method as described by Hosseini et al. (2015) with slightly modifica-
our previous methods (Wu et al., 2016).
tion. In brief, 1% (w/w) KGM solution was obtained by dissolving KGM
in hot distilled water (45 °C) under magnetic stirring for 6 h. CGNPs
were dispersed into distilled water and sonicated for 10 min, and the 2.5. Statistical analysis
suspensions were added dropwise to the KGM solution and gently
stirred for 2 h. The CSNPs content were 0%, 5%, 10%, 15% (w/w) on a Data were presented as the mean ± standard deviation of each
dry basis of the weight of KGM, and the corresponding resultant films treatment. Analysis of variance (ANOVA) was performed using a SPSS
were coded as KGM, KGM/CGNPs 5%, KGM/CGNPs 10%, KGM/CGNPs software statistical analysis system (SPSS 20.0 for windows, SPSS Inc.,
15%, respectively. Glycerol (0.1 g/g KGM) was added as a plasticiser in Chicago, IL). Duncan's multiple range test (p < 0.05) was used to
the solution in order to make the final films less brittle and easier to compare the differences amongst films.

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C. Wu et al.
Fig. 1. Schematic illustrations of the proposed mechanism of CGNPs formation.
3. Results and discussion
3.1. Characterization of CGNPs
CGNPs were obtained via an ionic cross-linking reaction between
the positively charged amino groups (-NH3+) of CS and the negatively
charged tripolyphosphate groups (-P3O105−) of TPP (Fig. 1)
(Bugnicourt & Ladavière, 2016; Wu et al., 2016). An ionic interaction
was established in the reaction between CS and TPP, thereby enabling
the encapsulation of an amount of GA within CSNPs. The morphological
characteristics of CGNPs were observed through TEM (Fig. 1), and the
individual CGNPs with an average size of 80–100 nm were spherical;
these findings were similar to previous result (Lamarra et al., 2016).
Dynamic light scattering (DLS) was also applied to investigate the
mean particle size and particle size distribution of the NPs. In Fig. 2a,
the size distribution profile of CGNPs had a mean diameter of
227.38 nm in a narrow size distribution (PDI = 0.185). The discrepancy
in the diameter of NPs through DLS and TEM could be attributed to the
mean hydrodynamic diameter of the particle core surrounded by sol-
vation layers as indicated by DLS which could not distinguish between
nanoparticle impurities and the sample; by contrast, the diameter of
particles alone in a dry state can be determined through TEM (Wu et al.,
2016).
According to DLVO theory (Verwey, 1947), particle aggregation can
be prevented by electrostatic repulsions, and a zeta potential of
+30 mV is required as a minimum for a physically stable nanosus-
pension solely stabilised by electrostatic repulsion. In Fig. 2b, the zeta
potential of CGNPs nanoparticles prepared in the present study was
about +59.2 mV, revealing highly stable suspensions. This positive zeta
potential was due to the cationic characteristics of CS chains in acetic
acid solution. This finding was consistent with those of Lamarra et al.
(2016), Bugnicourt and Ladavière (2016) and Wu, Hu, et al. (2016) and
Wu, Tian, et al. (2016).
3.2. Characterization of film-forming solutions
The rheological behaviour of film-forming solutions is an important
property that can indicate the relationships between the mechanical
behaviour and structure of biopolymer solutions (Liang et al., 2018).
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Food Hydrocolloids 89 (2019) 682–690
Therefore, the rheological properties of KGM-based film-forming solu-
tions should be investigated.
In Fig. 3a, the apparent viscosity (ηap) of all of the sample solutions
decreased as the shear rate increased, indicating the pseudoplastic
properties or the shear thinning region of these film-forming solutions.
A similar result has been observed in polysaccharide-based film-
forming solutions (Liang et al., 2018; Silva-Weiss, Bifani, Ihl, Sobral, &
Gómez-Guillén, 2014; Wu et al., 2016). ηap of KGM/CGNPs bionano-
composite film-forming solutions increased gradually compared with
that of the pure KGM solution as CGNPs loading increased from 5% to
10%, suggesting that the intermolecular interaction between KGM is
destroyed, and new hydrogen bonds are formed within the bionano-
composite film-forming solutions (El Miri et al., 2015). CGNPs could
also function as nanofillers that eliminated the free volume in a KGM
matrix network, leading to an increase in the viscous property of bio-
nanocomposite film-forming solutions. However, a further increase in
CGNPs loading (15%) in the KGM matrix resulted in a reduced ηap of
KGM/CGNPs film-forming solutions possibly because of the over-
loading or jamming of CGNPs within the KGM matrix. Therefore, 10%
CGNPs seemed to be the critical loading concentration of the KGM
matrix, thereby producing a reinforcement of the network.
The dynamic rheological properties of the film-forming solutions
were also determined via an oscillatory test to enhance our under-
standing of the delicate network structure of polymer systems. Angular
frequency sweep tests were also performed within the linear region
(about 1% of strain). The storage modulus (G′, represents elastic be-
haviour) and the loss modulus (G″, represents viscous behaviour) of the
film-forming solutions are presented in Fig. 3b.
G′ and G″ of all film-forming solutions increased as frequency in-
creased. G″ was higher than G′ at a low frequency, whereas G′ was
higher than G″ at a high frequency, indicating that all of the solutions
can be classified as weak gel systems (Long, Zhao, Zhao, Yang, & Liu,
2012). G′ and G″ of the KGM matrix increased after 5% CGNPs were
added, suggesting a well-dispersed CGNPs at the nanometric scale
within KGM matrix, and new hydrogen bonding interactions formed
between CGNPs and KGM, thereby enhancing the microstructure of the
film-forming solutions and increasing the mechanical strength of the
resulting films. The observation was also evident at a loading of 10%.
At this value, G′ and G″ significantly increased, and a solid-like flow
C. Wu et al.
Fig. 2. (a) Size distribution and (b) zeta potential distribution of CGNPs determined by DLS technique.
behaviour of the solutions was observed. It could be treated as an in-
terconnected gel-like network structure, and this assumption was con-
sistent with the expectation that CGNPs could cause additional inter-
facial interactions between their functionalised surfaces and the KGM
molecule chains. However, further addition of CGNPs (15%) into KGM
matrix led to a decrease in G′ as ηap decreased. Thus, 10% NPs could be
the critical concentration of the composite to develop a strong network
of CGNPs in a KGM matrix, and further loading of CGNPs substantially
weakened the network.
3.3. Characterization of bionanocomposite films
3.3.1. Film microstructure
The surfaces and cross-sections of KGM and KGM/CGNPs bionano-
composite films were observed under a SEM. As shown in Fig. 4a, the
pure KGM films displayed smooth and homogeneous surfaces, but small
pores or cracks appeared on the surface and cross-section. However, a
different superficial arrangement was observed after CGNPs were in-
corporated into the KGM film matrix (Fig. 4b–d). The incorporation of
CGNPs (5 wt.% and 10 wt.%) imparted a relatively compact and smooth
structure without any visible pores and cracks on the film's cross-sec-
tional microstructure. This might be related to the uniform dispersion of
CGNPs within the KGM matrix and their good miscibility and com-
patibility, thereby forming strong interaction and adhesion on the in-
terface between CGNPs and KGM chains. However, the surface became
less homogeneous with visible microcracks or discontinuous void when
15% CGNPs were incorporated into the KGM matrix. Compared with
the cross-sectional image of the KGM/CGNPs 10% film, visible pores
and cracks were observed in KGM/CGNPs 15% film. This phenomenon
would be attributed to the agglomeration of CGNPs in the KGM matrix
at high concentrations. The uniform dispersion of CGNPs within the
KGM matrix was directly correlated with its effectiveness in improving
the properties of bionanocomposite films. A similar microstructure has
been reported in polyvinyl alcohol, gelatin, pectin, tara gum and starch
film incorporated with CSNPs (Antoniou et al., 2015; Chang et al.,
2010; Hosseini, Rezaei, Zandi, & Farahmandghavi, 2016; Liu et al.,
2015; Lorevice et al., 2016; Ma, Liang, Cao, & Wang, 2018).
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Food Hydrocolloids 89 (2019) 682–690
The 3D surface morphology of KGM/CGNPs bionanocomposite films
was also analysed through AFM. In Fig. 5a, the pure KGM films had a
relatively homogeneous surface, with Ra and Rq of 10.5 and 13.7 nm,
respectively. The addition of CGNPs led to a remarkable increase in the
surface roughness of the bionanocomposite films as indicated by the
increase in Ra and Rq from 17.8 nm to 30.8 nm and from 20.2 nm to
45.7 nm, respectively (Fig. 5b–d). Several small sharp peaks were also
observed in the KGM/CGNPs bionanocomposite films probably because
of CGNPs. This result was consistent with those obtained from the SEM
micrographs. Likewise, Antoniou et al. (2015) reported a significant
increase in the surface roughness of tara gum/chitosan nanocomposite
film compared with pure tara gum films. Liu et al. (2015) also reported
an increase in the surface roughness of gelatin films after tea poly-
phenol-loaded CSNPs were added. Conversely, Hosseini et al. (2016)
reported that the surface roughness decreased when oregano essential
oil was added to fish gelatin/CSNPs composite films. This difference
may be due to the type of biopolymers, CSNPs content and biopolymer
affinity.
3.3.2. FT-IR and XRD analysis of films
The FT-IR spectra (Fig. 6a) were obtained to characterise the
functional groups of KGM/CGNPs bionanocomposite films and to
evaluate the interactions between KGM and CGNPs. In the FT-IR spectra
of the pure KGM film, the absorption bands at 3402, 2924, 1725 and
894 cm−1 were attributed to O–H stretching vibrations, C–H stretching,
C=O stretching vibrations and mannose unit stretching vibrations,
respectively. The absorption band at 1644 cm−1 was assigned to intra-
molecular hydrogen bonds. These findings were consistent with pre-
vious reports (Wu et al., 2012; Yuan et al., 2018). The spectrum of GA
showed typical phenolic characteristics with the existence of an O–H
stretching of a benzene ring within 3600–3100 cm−1, O–H plane
bending at 1365 cm−1, C=C stretching of an aromatic ring within
1450–1600 cm−1, and C–O/C–C stretching vibrations within
1200–1300 cm−1. In the spectrum of CGNPs, the main characteristic
peaks of CS at 3393 cm−1 (O-H stretching), 2899 cm−1 (C–H
stretching), 1640 cm−1 (amide I), 1550 cm−1 (amide II) and
1327 cm−1 (C–N stretching) were observed (Shah et al., 2016; Wu
C. Wu et al.
Fig. 3. Steady (a) and dynamic (b) rheological behaviour of bionanocomposite film-forming solutions.
et al., 2016). However, the O–H stretching vibration of the hydroxyl
groups of GA at 3494 cm−1 disappeared in the spectrum of CGNPs.
Similar changes in FT-IR spectra were also observed by Rosa et al.
(2013) when evaluating the encapsulation of gallic acid in chitosan
matrix. The spectrum of the bionanocomposite film incorporated with
10% (w/v) CGNPs showed the distinctive peaks of the pure KGM film
and the characteristic bands attributed to CGNPs. However, some of the
peaks shifted or disappeared with the incorporation of CGNPs. For in-
stance, the broad band around 3200–3600 cm−1 significantly enhanced
(the stretching vibration of O–H and N–H) and the peaks of amide II
(the characteristic peak of CGNPs) shifted to a higher wave number for
the KGM/CGNPs films, suggesting the formation of additional hydrogen
bonds between KGM and CGNPs in the bionanocomposite films. This
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Food Hydrocolloids 89 (2019) 682–690
might contribute to a good compatibility between KGM and CGNPs
components, resulting in enhanced physicochemical and mechanical
properties of bionanocomposite films.
XRD was performed to verify the compatibility of KGM and CGNPs
in the bionanocomposite films (Fig. 6b). A broad peak at 2θ = 22.6°
was observed in the pure KGM film, indicating that KGM is an amor-
phous material which was consistent with previous findings (Li et al.,
2015; Yuan et al., 2018). The X-ray patterns of all of the bionano-
composite films were similar to those of the pure KGM film, suggesting
the good compatibility of KGM and CGNPs in the bionanocomposite
films. These results supported the results of FT-IR analysis.
C. Wu et al.
Fig. 4. The surface and cross-section microstructures of (a) KGM films, (b) KGM/CGNPs 5% films (c) KGM/CGNPs 10% films, and (d) KGM/CGNPs 15% films.
3.3.3. Thermal properties of bionanocomposite films
The thermal properties of the bionanocomposite films are shown in
Fig. 6c. According to the TGA curves, the behaviour of all of the films
was similar to the two main stages of thermal degradation at 25 °C-
600 °C. The first mass loss of all of the samples occurred at around
100 °C because of the loss of adsorbed water (Neo et al., 2013). The
second mass loss between 250 °C and 450 °C was due to the thermal
decomposition of KGM and CGNPs. The mass loss of the bionano-
composite films at the tested temperature was less than that of the pure
KGM films, indicating that the incorporation of CGNPs enhanced the
thermal stability of the bionanocomposite film. The thermal stability of
the bionanocomposite films increased as CGNPs increased from 0% to
10% and then decreased at 15%. The thermal stability of the biona-
nocomposite film with more than 10% CGNPs slightly decreased likely
because of the aggregation of CGNPs in the bionanocomposite matrix.
3.3.4. Transparency properties of bionanocomposite films
Film transparency is an important parameter of general appearance
and consumer acceptance. The transmission of UV and visible light at
the selected wavelength (200–600 nm) of the films is shown in Fig. 7. In
the UV range of 200–400 nm, the KGM/CGNP bionanocomposite films
exhibited excellent UV shielding properties compared with those of the
pure KGM film, possibly protecting foods from oxidative deterioration,
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Food Hydrocolloids 89 (2019) 682–690
nutrient loss, discolouration and off flavours (Ma, Du, Yang, & Wang,
2017). A similar behaviour has also been observed in polysaccharide-
based films containing natural extracts rich in phenolic compounds
(Vilela et al., 2017), and protection against UV light is increased by
increasing the content of active components. In the visible light region
(400–600 nm), the pure KGM exhibited a good optical transmittance of
88%–93%. The optical transmittance properties of the bionanocompo-
site films were not greatly influenced by the addition of CGNPs into the
KGM matrix. This finding confirmed that CGNPs were well compatible
with KGM and well dispersed in the bionanocomposite films. The ex-
cellent transparency properties were also supported by the visual ob-
servations of bionanocomposite films (Fig. 7). The plant covered with
the bionanocomposite films could be observed clearly, indicating that
KGM/CGNPs bionanocomposite films with good transparency and UV
barrier properties showed potential for packaging applications.
3.3.5. WVP of the films
WVP of the pure KGM film was 18.22 × 10−11 g/(m*s*Pa) which
was significantly decreased by the addition of CGNPs into the KGM
matrix (p < 0.05) as shown in Fig. 8. KGM is highly sensitive to
moisture and has poor WVP properties because of its hydrophilic
character (Li et al., 2015). Increasing the amount of cross-links via
hydrogen bonding between CGNPs and KGM might form a film network
C. Wu et al. Food Hydrocolloids 89 (2019) 682–690

Fig. 5. AFM micrographs of (a) KGM films, (b) KGM/CGNPs 5% films (c) KGM/CGNPs 10% films, and (d) KGM/CGNPs 15% films.

with the reduced free volume of the film matrix, leading to a compact
structure of the bionanocomposite film and further decreasing the WVP
of the resulting films. However, WVP did not significantly differ
(p > 0.05) between the films incorporated with the addition of 10%
and 15% w/w CGNPs in the bionanocomposite films. This phenomenon
might be due to the overloading of CGNPs (15%) in the KGM matrix,
resulting to the decrease of the compactness structure of KGM/CGNPs
film. Hosseini et al. (2016) also found a decreased WVP of fish gelatin-
based edible films incorporated with CS/oregano essential oil NPs.
Other studies have also revealed a decrease in the WVP of poly-
saccharide-based films with chitosan NPs (Antoniou et al., 2015; Chang
et al., 2010; Moura, Lorevice, Mattoso, & Zucolotto, 2011; Lorevice

Fig. 6. FT-IR, XRD and TGA spectra of bionanocomposite films.

688
C. Wu et al.
Fig. 7. Light transmission characteristics and physical appearance of films.
Fig. 8. WVP (a) and mechanical properties (b) of KGM/CGNPs bionano-
composite films.
et al., 2016).
3.3.6. Mechanical properties of the films
The mechanical properties, including TS and EB, of the bionano-
composite films are presented in Fig. 8b. TS of the bionanocomposite
films was higher than that of the pure KGM film, whereas EB of the
former was lower than that of the latter. The TS of bionanocomposite
films was significantly enhanced by the increased CGNPs loading con-
tent from 5% to 10% (p < 0.05) and decreased slightly thereafter. This
phenomenon could be attributed to the homogeneous dispersion of
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Food Hydrocolloids 89 (2019) 682–690
CGNPs in the KGM matrix, thereby forming a new hydrogen bond be-
tween the filler (CGNPs) and the matrix (KGM). As a result, the struc-
ture network was reinforced as indicated by FT-IR, SEM, AFM and
rheological analysis. However, the overloading of CGNPs (15%) re-
sulted in the inhomogeneous distribution of CGNPs in the KGM matrix
and slightly decreased the mechanical property (p > 0.05). EB of the
bionanocomposite films decreased as the NPs concentration increased
(p < 0.05). EB decreased from 42.23 ± 4.42% to 26.61 ± 3.28% by
increasing the content of CGNPs. The decreased EB could be attributed
to the improvement in the rigidity of bionanocomposite films. The
addition of CGNPs filled in the free volume between polymer chains
caused by an increase in intermolecular attractive forces, thereby
making the polymer network highly dense and less permeable (Hosseini
et al., 2015). Therefore, direct interaction and proximity between
polysaccharide chains decrease, thereby reducing EB. A scheme illus-
trating the strengthening mechanism of CGNPs on the physico-chemical
properties of bionanocomposite films is presented in Fig. 8. A similar
result between a nanofiller and a polysaccharide matrix in the biona-
nocomposite films has been reported (Antoniou et al., 2015; Chang
et al., 2010; Moura et al., 2011; Lorevice et al., 2016).
3.3.7. Antibacterial properties of bionanocomposite films
The antibacterial activities of pure KGM films and KGM/CSNPs
bionanocomposite films against gram-positive S. aureus and gram-ne-
gative E. coli are illustrated in Fig. 9. The pure KGM films did not show
any antibacterial activity against both gram-positive and gram-negative
bacteria. On the contrary, KGM/CSNPs bionanocomposite films ex-
hibited slightly different CGNPs concentration-dependent antibacterial
activities against E. coli and S. aureus strains from 5% to 15%. The
antibacterial activity of the bionanocomposite films against E. coli was
weaker than that against S. aureus possibly because of the difference in
cell wall structures between gram-positive and gram-negative micro-
organisms. CS's antimicrobial activity is higher against gram-positive
bacteria than against gram-negative bacteria (Zou et al., 2016). GA can
also disrupt peptidoglycan or disintegrate the outer membranes of
bacteria through metal ion chelation, leading to bacterial cell death
(Lee & Je, 2013; Neo et al., 2013). Although the aggregation or ag-
glomeration of CGNPs (15% w/w overloading) decreased the specific
surface area, the increasing GA content would improve their anti-
microbial activity. Thus, the antimicrobial activity of the bionano-
composite films was enhanced by the increasing amount of CGNPs.
Similar observations were obtained by Antoniou et al. (2015) and
Hosseini et al. (2015) in tara gum- and gelatin-based films with in-
corporated CSNPs and CSNPs/oregano essential oil, respectively. This
result suggested that KGM/CGNPs bionanocomposite films in-
corporated with 10% CGNPs could act as antimicrobial films against
Fig. 9. Antibacterial activity of KGM/CGNPs bionanocomposite films.
C. Wu et al.
microorganisms.
4. Conclusion
An active bionanocomposite composed of KGM and CGNPs was
successfully prepared by solvent casting. CGNPs with a GA loading ef-
ficiency of 78.0 ± 2.3% were obtained by ionotropic gelation method.
The resultant CGNPs were spherical with a diameter range of
80–100 nm. Consistent with FT-IR, XRD and TGA results, the rheolo-
gical results of film-forming solutions revealed that CGNPs interacted
with KGM through hydrogen bonds in the bionanocomposite matrix.
The microstructure of the films showed that the introduced 5–10%
CGNPs appeared to be homogeneously dispersed within the KGM film
matrix, thereby reducing the free volume of the composite matrix and
significantly improving the mechanical, barrier and thermal properties
of the final bionanocomposite film (p < 0.5). The incorporation of
CGNPs to the KGM matrix improved the UV barrier abilities and anti-
microbial properties of the resultant bionanocomposite. Therefore,
KGM/CGNPs bionanocomposite films showed promising potential as
fruit packaging material in food industries.
Acknowledgements
This work was supported by the National Science Foundation of
China (No. 31801616 and 31471704).
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