Food Chemistry 430 (2024) 137033

Contents lists available at ScienceDirect

Food Chemistry
journal homepage: www.elsevier.com/locate/foodchem

Effects of glycation method on the emulsifying performance and interfacial

behavior of coconut globulins-fucoidan complexes
Qianqian Zhu a, Haiming Chen a, Weijun Chen a, Ming Zhang a, Qiuping Zhong a, Zixin Chen b,
Jianfei Pei a, *, Wenxue Chen a, *
a
School of Food Science and Engineering, Hainan University, 58 People Road, Haikou 570228, PR China
b
Wenchang Zaineng Industrial Co., Ltd., Dongjiao Town, Wenchang 571300, PR China

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

Keywords: Coconut globulins (CG) possesses potential as an emulsifier but has not been utilized well. In this study, the
Coconut emulsifying performance of glycated CG-fucoidan (CGF) complexes, and the relationship between emulsifying
Globulins stability and interfacial behavior were investigated. The results showed that the grafting of fucoidan increased
Fucoidan
the molecular weight of CG, and decreased the zeta potential and fluorescence intensity. With the higher
Glycosylation
glycosylation degree, the fucoidan modified CG exhibited better emulsifying stability and higher viscosity.
Emulsion
Interfacial adsorption Moreover, the result of adsorption kinetics revealed that elasticity was the main property of the interface layer.
Compared to CG, CGF complexes with high degree of glycosylation had thicker interfacial layer on the oil–water
interface. A thicker elastic interfacial layer may be beneficial to the emulsion stability, owing to the strong
interaction of electrostatic repulsion and steric hindrance between oil droplets. These findings may provide
useful information for glycated CGF complexes as emulsifiers in functional food.

1. Introduction
Plant-based proteins are of increasing interest to scientists as emul­
sifier of multiphase food system, owing to their advantages of nutritional
and functional properties (Drusch et al., 2021). Coconut protein can
quickly adsorb on the oil–water interface to reduce interfacial tension. It
seems to be a promising emulsifier alternative with a well-balanced
amino acids profile, the compositions contain all the nine essential
amino acids, and non-essential amino acids (Martinez-Padilla et al.,
2022). The amount of essential amino acid contains 71% to 77%, and it
can be easily digested and absorbed by the body (Patil & Benjakul,
2018). However, coconut protein is usually discarded in the environ­
ment as a waste by-product after the extraction of virgin coconut oil
(Thaiphanit & Anprung, 2016). Although coconut protein only makes up
2.6–4.4% (w/w) of fresh coconut meat, the production is very large
(Patil & Benjakul, 2018; Thaiphanit & Anprung, 2016). As reported by
FAO, the production of coconut in 2021 was about 63.68 million tonnes
globally (https://www.fao.org). The present treatment may not only
lead to the squandering of resource, but also contaminate the environ­
ment. How to implement the high-value utilization of coconut protein is
a problem that needs to be solved.
Currently, it is an attractive research direction to develop coconut
protein as an emulsifier for high-value utilization in the food industry.
Coconut proteins were separated by different solvents, mainly contained
albumins, globulins, gliadins, glutelins-1 and glutelins-2, of which
globulins accounted for 60–75% of the total protein isolates (Patil &
Benjakul, 2017). The majority of coconut globulins (CG) is the 11S
globulins, also called cocosin. It has been reported that CG is a hexamer
of 55 kDa subunit linked by disulfide bonds, the subunit is consisted of
an acidic polypeptide and a basic polypeptide (Ma et al., 2022). Previous
studies have proved that CG has excellent emulsifying ability and can
quickly adsorbed on the oil–water interface to reduce interfacial tension
(Patil & Benjakul, 2017). However, the solubility of CG is so low that its
emulsifying stability is poor, which limits its applications in food in­
dustry (Raghavendra & Raghavarao, 2012). There are many modifica­
tion methods used for promoting properties of plant-based proteins,

Abbreviations: CG, Coconut globulins; CGF, Coconut globulins-fucoidan; CI, Creaming index; DG, The degree of grafting; FTIR, Fourier-transform infrared
spectroscopy; OPA, O-Phthalaldehyde; QCM-D, Dissipative quartz crystal microbalance; SDS-PAGE, Sodium dodecyl sulfate–polyacrylamide gel electrophoresis;
UV–vis spectrophotometer, Ultraviolet–visible spectrophotometer.
* Corresponding authors at: School of Food Science and Engineering, Hainan University, 58 People Road, Haikou 570228, PR China.
E-mail addresses: peijianfei@hainanu.edu.cn (J. Pei), chwx@hainanu.edu.cn (W. Chen).

https://doi.org/10.1016/j.foodchem.2023.137033
Received 4 May 2023; Received in revised form 20 July 2023; Accepted 25 July 2023
Available online 26 July 2023
0308-8146/© 2023 Elsevier Ltd. All rights reserved.
Q. Zhu et al.
such as ultrasonic treatment, enzymatic hydrolysis and glycosylation
(Ding et al., 2021; Lad & Murthy, 2012; Li et al., 2022). Glycosylation is
a relatively safe, simple method to improve the functional properties of
protein, which can stabilize emulsions by regulating the thickness of
interfacial layer (Ai et al., 2021). The concept has been studied in
emulsion stability, the encapsulation of bioactive substance and pro­
tection against oxidation (Zhang et al., 2014; Zhang et al., 2022). At
present, studies on the emulsifying performance of glycated CG are
poorly reported. Moreover, the mechanism, especially the relationship
between emulsifying stability and thickness of interfacial layer formed
by CG-polysaccharide was not studied.
Fucoidan is a sulfated anionic polysaccharide extracted from brown
algae and echinoderms, mainly composed of repeating fucose unit with
sulfated ester groups. Fucoidan has excellent health benefits and phar­
macological activity, such as anti-inflammatory, antitumor and antiox­
idant activity (Liu et al., 2020). Many bioactive properties of fucoidan
comes from the presence of sulfated groups (Alghazwi et al., 2019). The
anionic groups of fucoidan has the potential to interact with cationic
amino acid residues on CG, thereby improving the physicochemical and
emulsifying properties (Liu et al., 2020).
Therefore, the present study focused on the globulins in coconut, the
emulsifying stability of glycated coconut globulins by fucoidan, and the
relationship of emulsifying stability and interfacial adsorption behavior
were investigated. First, fucoidan was conjugated on the CG using wet-
heating method. The structural characteristics of CGF were analyzed by
grafting degree, protein pattern, FTIR, fluorescence spectroscopy and
zeta potential. Then, the emulsifying performance of CG and glycated
CGF complexes was studied by particle size, morphology, viscosity and
centrifugal stability. Finally, the adsorption behavior of glycated CGF
complexes on the oil–water interface was evaluated using interfacial
dilatational rheology and dissipative quartz crystal microbalance. In
addition, the emulsifying stability-interfacial behavior relationship of
glycated CGF complexes were proposed.
2. Materials and methods
2.1. Materials
The by-products of coconut skim milk without heat treatment was
kindly provided by Wenchang Zaineng Industrial Co., Ltd (Wenchang,
China). Fucoidan (Mw = 190 kDa) was purchased from Macklin
Biochemical Co., Ltd. (Shanghai, China). Medium-chain triglyceride
(MCT) was supplied by the Nisshin Oillio Group (Tokyo, Japan). O-
Phthalaldehyde, KBr were purchased from Sangon Biotech Co., Ltd.
(Shanghai, China). Other reagents were analytical grade.
2.2. Preparation glycated CGF complexes
2.2.1. Purification of coconut globulins
Purification of coconut globulins was carried out according to the
modified method of Ma et al. (2023). The by-product of coconut skim
milk was dispersed in deionized water with a ratio of 1:20 (w/v), and
then stirred at 4℃ for 2 h. The mixture was centrifuged at 10000 rpm for
15 mins, and there were three phases in the system: cream, aqueous
solution with albumins and precipitation. The precipitation was
collected to extract globulins using 0.4 M NaCl in 35 mM sodium
phosphate buffer (pH 7.0). Extraction procedure with deionized water
and NaCl solution were repeated three times. The supernatant was
collected and dialyzed with deionized water for 48 h (6–8 kDa molecular
weight cut off). The dialysate was freeze-dried and stored at − 20℃ for
further analysis.
2.2.2. Preparation of CGF complexes by Millard reaction
Glycated CGF complexes were prepared following the steps below:
(i) The CG (2.0 g) was dissolved in 0.4 M NaCl solution in 35 mM
potassium phosphate buffer (200 mL, pH 7.0) to obtain a CG stock
2
Food Chemistry 430 (2024) 137033
solution (10 mg/mL). Different quantities of fucoidan (0.1, 0.5, 1.0, 2.0,
3.0 and 4.0 g) were dissolved in 100 mL deionized water to obtain
fucoidan stock solution (1, 5, 10, 20, 30, 40 mg/mL).
(ii) To prepare glycated CGF complexes, 20 mL of CG stock solution
was mixed with 20 mL fucoidan stock solution. The Millard reaction was
performed at 60℃ for 2 h, based on the method of Zhang et al. (2022)
with some modification. CG with or without heat treatment at 60℃ were
used as a control group,
After the Millard reaction, the samples were immediately transferred
to ice bath to terminate the reaction. One part was used to measure the
degree of grafting, SDS-PAGE, fluorescence spectroscopy, the other part
was freeze-dried at − 50℃ for emulsion preparation and interfacial
behavior study.
In this study, glycated CGF complexes with different weight ratios of
10:1, 2:1, 1:1, 1:2, 1:3, 1:4 were named CGF10:1, CGF2:1, CGF1:1, CGF1:2,
CGF1:3, CGF1:4 respectively.
2.3. Characterization of the degree of glycosylation and structure of CGF
complex
2.3.1. Degree of grafting
The grafting degree (DG) of CGF complexes was measured according
to OPA method (Feng et al., 2023). The OPA solution was prepared as
follows, 40 mg O-Phthalaldehyde (OPA) was dissolved in 1 mL methanol
and then mixed with 2.5 mL SDS (20%, w/v), 25 mL sodium borate
solution (0.1 mol/L) and 100 µL β-mercaptoethanol. The final volume
was 50 mL. Next, 200 µL of the CG or CGF complexes sample was added
to 4 mL of OPA solution, and incubated at 35℃ for 2 mins. The absor­
bance was determined at 340 nm using UV–vis spectrophotometer
(Mettler Toledo, Switzerland). The DG was calculated by following
equation:
A0 − At
DG (%) = × 100% (1)
A0
A0 and At were the absorbance before and after the Maillard reaction.
2.3.2. Sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-
PAGE)
The SDS-PAGE experiment was performed according to Gerliani’s
method with some modification (Gerliani et al., 2019). The concentra­
tion of separating gel was 12%, and that of stacking gel was 4%. The
samples at 2 mg/mL were added to loading buffer with a ratio of 4:1 and
boiled for 10 mins before loading to gel. The voltage of running stacking
gel was 80 V, and running the separating gel at 120 V. The gel was
stained with Coomassie brilliant blue R250. The molecular weight of CG
and CGF complexes were calculated using a standard protein marker
(10–170 kDa).
2.3.3. Zeta potential
Zeta potential was measured by Zetasizer (Malvern Instrument, UK)
as reported (Zhao et al., 2023). The samples were diluted 10-fold with
35 mM sodium phosphate buffer (pH 7.0) and then added into the
sample cell. All measurements were tested in triplicate at 25℃.
2.3.4. Fourier-transform infrared (FTIR) spectroscopy
The FTIR spectra of CG and glycated CGF complexes were acquired
using FTIR spectroscopy (Bruker Corp., Germany). At first, the sample
was grinded with KBr power under the ratio of 1:100 (w/w), and then
tableted. The spectra were scanned in the range of 4000–400 cm− 1 with
a resolution of 4 cm− 1 (Li et al., 2022).
2.3.5. Fluorescence spectroscopy
The fluorescence spectroscopy of CG and CGF complexes were
recorded by F-6000 fluorescence spectrophotometer (Hitachi, Japan).
The excitation wavelength was set to 280 nm, and emission wavelength
was 290–500 nm (Xia et al., 2015). The slit width was 5 nm.
Q. Zhu et al.
2.4. Emulsifying properties of CGF complexes
2.4.1. Emulsions preparation
Different glycated CGF samples (CGF10:1, CGF2:1, CGF1:1, CGF1:2,
CGF1:3, CGF1:4) and CG with or without heat treatment (1%, w/v) were
hydrated overnight, then 20 wt% MCT oil was added to the solution. The
mixture was sheared at 12000 rpm for 2 mins by a high speed disperser
(IKA T18 digital ULTRA-TURRAX, Germany) to prepare emulsions (Ma
et al., 2023).
2.4.2. Droplet size and CI index of emulsions
The droplet size of emulsions was measured by a static laser light
scattering instrument (Malvern Instruments, UK). Fresh emulsion was
added to the sink and dispersed automatically until the appropriate laser
obscuration. The refractive index of MCT oil and water were set to 1.45
and 1.33, respectively. The surface-weighted mean diameter (d3,2) and
volume-weighted mean diameter (d4,3) were recorded (Niu et al.,
2022b).
Creaming index (CI) index was calculated as follows: CI% = (height
of the cream layer / total height of the emulsion) × 100%.
2.4.3. Fluorescence micrograph of emulsions
The microstructure of emulsion was observed by inverted fluores­
cence system (MODEL ECLIPSE Cl-S, OPTIKA, Italy). Adding 40 µL Nile
red solution (0.2 wt%, dissolved in isopropanol) into 1 mL fresh emul­
sion, mixed and stained at dark for 2 h. The sample was distributed
across a microslide, and immediately covered with a cover slip, imaged
at 10 × magnification (Ma et al., 2023).
2.4.4. Viscosity measurement
Apparent viscosity of emulsions was determined by HAAKE MARS III
rotary rheometer (ThermoFisher, USA). The emulsion was added to the
parallel plate, and scraped off the excess sample outside the plate. The
gap of measurement was set to 1.000 mm. The apparent viscosity of
emulsions was measured at the shear rate in the range of 1–100 s− 1. All
measurements were performed at 25℃ (Chen et al., 2023).
2.4.5. Long-term stability measurement
The long-term stability of emulsions was evaluated using LUMiSizer
stability analyzer (Berlin, Germany). About 400 µL fresh emulsion was
added to the bottom of the sample cell, and then placed horizontally into
the instrument. According to the standard procedure, the wavelength
was 870 nm, and emulsions were centrifuged at 4000 rpm for 50 mins.
All measurements were taken at 25℃. The transmission profile was
recorded with time evolution. Integral transmission and instability index
were calculated by SEPView 6.0 software after measurement (Niu et al.,
2022b).
2.5. Dilatational rheology measurement
2.5.1. Dynamic interfacial tension and interfacial dilatational rheology
Dilatational rheology measurement was performed at an optical
contact anglemeter (Dataphysics Instruments GmbH, Germany) ac­
cording to a previously established procedure with some modification
(Niu et al., 2022a). The droplet volume on the test of dynamic interfacial
tension was 25 µL. The interfacial pressure (π, mN/m) was calculated by
the equation as follows (Yang et al., 2023):
π = γ0 − γ (2)
where γ0 is the interfacial tension of buffer solution and MCT oil
(20.35 mN/m), and γ represents the interfacial tension of CG and CGF
complexes with MCT oil.
The diffusion rate of CG and CGF at oil–water interface was analyzed
by Ward and Tordai equation:
π = 2C0 KT(Dt/3.14)1/2 (3)
3
Food Chemistry 430 (2024) 137033
where C0 is the concentration of the continuous phase, K is the
Boltzman constant, T is the absolute temperature, t is the adsorption
time, and D is the diffusion coefficient.
Furthermore, the penetration and rearrangement rate of CG and CGF
at O/W interface were analyzed by the first-order equation:
[( )/( )]
ln πf − π t π f − π 0 = − ki t (4)
where πf, π0, πt represent the interfacial pressure at the final
adsorption time (10800 s), at the initial time and t time respectively.
The interfacial viscoelastic modulus of CG and CGF solutions (15 µL)
at the O/W interface was measured, the deformation amplitude (ΔA/A)
was set to 10% at 0.1 Hz frequency for 3 h. All measurements were
carried out at 25℃. tan φ was calculated according to following Eqs:
tanφ = Ev /Ed (5)
where φ represents phase angle between stress and strain. Ed is the
interfacial elastic modulus, and Ev is the interfacial viscous modulus.
2.5.2. Amplitude sweep
The deformation amplitude (ΔA/A) from 2.5% to 30% was set to
study the perturbation of amplitude sweep on the structure of interfacial
layer, the frequency was set to 0.1 Hz, and the droplet volume was 15 µL.
The interface was balanced for 3 h before measurement. To intuitively
show the structure change of interface caused by drop deformation, the
Lissajous plots were created (Ma et al., 2023).
2.5.3. Frequency sweep
The structure change of interface layer under frequency perturbation
was measured from 0.005 to 1 Hz. The droplet volume was 15 µL, and
deformation amplitude (ΔA/A) was 10%. The interface on oil–water was
balanced for 10800 s before measurement (Niu et al., 2022b).
2.6. QCM-D analysis
The adsorption behavior of CG and CGF on the oil–water interface
were studied by dissipative quartz crystal microbalance (QCM-D) (Biolin
Scientific AB, Sweden). Frequency and dissipation signal were recorded
simultaneously. According to previously established procedure (Niu
et al., 2022a), 300 µL MCT oil (0.5%, dissolved in chloroform) was
coated on the gold quartz sensor (QSX 301, 5 MHz), then the sensor was
dried overnight in a vaccum drying oven at 40℃. At experiment, the
sensors were cleaned and calibrated the baseline using buffer solution
firstly, and then the sample solutions were pumped into the measure­
ment cell by a peristaltic pump, the flow rate was set to 50 µL/min. After
adsorption equilibrium, the sensors were rinsed by buffer solution to
remove the unbound samples. The data was calculated by the smartfit
model of Qsense Dfind. The frequency and dissipation shifts were based
on the 3rd and 3th overtones.
2.7. Molecular docking
The interaction between CG and fucoidan was simulated by molec­
ular docking method (Feng et al., 2023). The main globular protein
(cocosin) in CG and the oligosaccharide structure simplified from
fucoidan were chosen to build the CGF conjugated structure. Crystal
structure of cocosin (5WPW) was downloaded from the Protein Data
Bank (https://www.rcsb.org/). The oligosaccharide structure of fucoi­
dan was mapped by the software of ChemOffice. First, we used PyMOL
to remove water molecules and add hydrogen atoms on cocosin struc­
ture, and then the protein and oligosaccharide unit structure were
further modified by AutoDock Tools 1.5.6 software. The molecular
docking was launched in AutoGrid and AutoDock. After 3D molecular
docking procedure was finished, the pdbqt format file was imported in
LigPlot software to generate 2D graph of CGF molecular docking map.
Q. Zhu et al.
Fig. 1. The grafting degree and SDS-PAGE pattern (A), Zeta potential (B), FTIR spectroscopy (C) and fluorescence emission spectra (D) of CG and glycated CGF
complexes. Different letters in each column indicate significant differences (P < 0.05).
2.8. Statistical analysis
All the experiments were conducted in triplicate, and the data was
expressed as average values ± standard deviations. Statistical differ­
ences between the experimental groups were calculated using the least
significant difference (LSD) test (P < 0.05) by Origin 2023 software (EI-
Hadary et al., 2023).
3. Results and discussion
3.1. Characterize glycated CGF complexes
3.1.1. Glycation degree of CG with fucoidan
A train of glycated CGF samples with different mass ratios (10:1, 2:1,
1:1, 1:2, 1:3, 1:4) was prepared. The characterization of glycated CGF
complexes was shown in Fig. 1, including the degree of grafting (DG),
SDS-PAGE pattern, Zeta potential, FTIR and fluorescence emission
spectra. The glycosylation of CG and fucoidan depends on the covalent
reaction of the amino groups of protein with the carbonyl groups of the
reducing sugar (Ai et al., 2021). Therefore, DG can be evaluated by
measuring the amount of free amino groups in the system. Fig. 1A de­
picts the DG values of glycated samples with different mass ratio of
4
Food Chemistry 430 (2024) 137033
fucoidan. The DG of glycated CGF complexes increased gradually as the
ratio of fucoidan increased (P < 0.05), and reached the maximum at a
ratio of 1:4, which was 55.70%. DG could be affected by the properties
of proteins and polysaccharides (Zhao et al., 2023), it reported that the
DG of apple pectin, citrus pectin, mango pectin, sugar beet pectin and
coconut protein isolate were 59.11%, 52.80%, 41.39%, and 39.2%
respectively (Li et al., 2022). The differences of DG maybe due to
different molecular weight of polysaccharide. Generally, the lower
molecular weight will cause the greater degree of glycosylation at the
same reaction condition.
SDS-PAGE was used to evaluated the glycosylation degree of CG and
fucoidan. As reviewed previous study, CG are composed of 11S globulins
or cocosin (86%), and 7S globulins (14%). Cocosin is a hexamer of 55
kDa subunit linked by disulfide bonds. The 7S globulins is a trimeric
protein comprised by 16, 22 and 24 kDa subunits (Ma et al., 2022). As
shown in Fig. 1A, the main bands of CG were mainly distributed in
15–55 kDa, a clear 11S globulins subunit was found in the lane of un­
treated CG and CG-heated, which was consistence with the results of Ma
et al. (2022). For the glycated CGF complexes, the intensity of 55 kDa
band decreased gradually with increasing the DG. Compared with CG or
CG-heated, there were obviously darker bands at the top of the stocking
gel and separation gel, which may be attributed to the conjugation of CG
Q. Zhu et al.
and fucoidan. With the increase of fucoidan ratio, the band of 55 kDa
gradually diminished, indicating the increase of glycation degree. The
SDS-PAGE pattern was consistent with the result of DG. Protein pattern
of glycated coconut protein isolates with pectin also exhibited a similar
phenomenon (Li et al., 2022).
3.1.2. Zeta potential
The zeta potential was used to measure the surface charge density of
particles, which could offer an indication of electrostatic repulsion
interaction. Fig. 1B shows the zeta potential of CG and CGF complexes
after glycosylation. The higher absolute value of zeta potential can in­
crease the electrostatic repulsion interaction, which lead to the greater
distance between particles. On the contrary, particles will tend to
coagulation or flocculation. The zeta potential of CG without treatment
was − 14.4 mV, and the CG-heated was − 11.0 mV, which reflected the
solution aggregation after heat treatment to some extent. The absolute
value of zeta potential of CG heated at 80℃ has also been reported to be
smaller than the control, the electrostatic repulsion interactions weak­
ened (Ma et al., 2022). When the value of DG and fucoidan ratio
increased, the absolute value of zeta potential increased significantly,
which was attributed to the increase of fucoidan grafting on the CG
surface. It is generally considered that when zeta potential value is
greater than ± 30 mV, the system is more stable (Lu et al., 2023).
Therefore, glycated CGF complexes with ratios of 1:1, 1:2, 1:3 and 1:4
were more stable with zeta potentials of − 35.53 ± 1.79 mV, − 46.03 ±
2.51 mV, − 49.93 ± 4.31 mV and − 46.63 ± 5.20 mV, respectively.
3.1.3. Structural characteristics of CGF
FTIR spectra reflect the secondary structure and hydrogen bonding
forces of protein, which is widely used to analyze the glycosylation
modification of protein. To verify the conjugate formation of CG and
fucoidan, FTIR analysis was used to reveal the characteristics of the
modified proteins. As shown in Fig. 1C, untreated CG showed typical
diffraction peaks at wavenumber of 3437.9, 1648.3, 1157.5, 863.8
cm− 1, that represented the O–H stretching vibrational, amide I bands
(C–– O stretching vibrational), NH2 inward osway and bending vibration
of NH groups, respectively. These bands are consistent with the reported
spectrum of coconut protein isolate (Li et al., 2022). There was not
obvious change in FTIR spectrum between CG-heated and natural CG.
After glycosylation modification, a new diffraction peak appeared at the
wavenumber of 1422.8 cm− 1, which represented the formation of C–N
covalent bond at the CG to fucoidan mass ratios of 10:1, 2:1, 1:1, 1:2,
1:3, 1:4 (Su et al., 2010). This was a typical characteristic of carbonyl­
amine binding between protein and carbohydrate by Millard reaction,
which was consistent with the phenomenon of canola protein isolate-
gum Arabic Maillard conjugate (Pirestani et al., 2018).
The fluorescence of aromatic amino acid residues is very sensitive to
surrounding environment (Li et al., 2019). Conformational changes of
CG modified by grafting fucoidan can be revealed by fluorescence
quenching experiment. The emission spectra of CG and CGF are shown
in Fig. 1D, CG exhibited the maximum emission peak at 339.6 nm when
stimulated by 280 nm excitation light. After glycosylation, the intensity
of fluorescence decreased with the increase in fucoidan ratio, which was
attributed to the shielding effect of polysaccharide chain (Fan et al.,
2017). For the covalent grafting between CG and fucoidan, carbonyl
groups could act as quenching agents causing the fluorescence
quenching. Zhang et al. (2022) also showed that glycated egg protein-
based complexes caused the fluorescence quenching phenomenon,
when conjugated with lactose, fructose, xylose and glucose.
3.2. Emulsifying properties of CGF complexes
3.2.1. Droplet size and microstructure of emulsions
Droplet size is one of important indicators to evaluate the stability of
emulsions. The effect of different concentration of fucoidan glycated CG
on the droplet size (d3,2 and d4,3) and CI index were studied in the O/W
5
Food Chemistry 430 (2024) 137033
Table 1
Droplet size and CI index of emulsion stabilized by CG and glycated CGF
complexes.
Sample Stored for 24 h
d3,2 (µm) d4,3 (µm) CI (%)
CG 2.88 ± 0.07a 13.43 ± 0.73a 19.27 ± 0.17a
CG-heated 2.95 ± 0.04a 13.48 ± 0.41a 18.35 ± 0.10b
CGF10:1 2.49 ± 0.33b 10.09 ± 2.89bcd 17.50 ± 0.13c
CGF2:1 2.34 ± 0.01b 8.73 ± 0.06d 15.34 ± 0.11e
CGF1:1 2.37 ± 0.01b 8.90 ± 0.04 cd 16.35 ± 0.13d
CGF1:2 2.77 ± 0.03a 10.60 ± 0.17bc ND
CGF1:3 2.82 ± 0.02a 10.35 ± 0.13bcd ND
CGF1:4 2.88 ± 0.04a 11.29 ± 0.17b ND
All data are expressed as mean ± standard deviation, and different letters
indicate significant difference (P < 0.05). ND: not detected.
emulsions. As shown in Table 1, the CI index of emulsions prepared by
CGF1:2, CGF1:3, CGF1:4 could not be detected after storage for 24 h,
which were relatively stable. The mean size of emulsion stabilized by CG
was 2.88 ± 0.07 µm. As the ratio of fucoidan gradually increased (from
10:1 to 1:1), the droplet size of oil was significantly (P < 0.05) decreased
to 2.49 ± 0.33 µm (CGF10:1), 2.34 ± 0.01 µm (CGF2:1) and 2.37 ± 0.01
µm (CGF1:1). This was due to the glycated CGF complexes could quickly
adsorb on the surface of oil droplets during high speed dispersion, and
formed a viscoelastic film to inhibit the small oil droplets from merging
into larger droplets (Niu et al., 2023). Interestingly, the size of oil
droplets increased to 2.77 ± 0.03 µm (CGF1:2), 2.82 ± 0.02 µm (CGF1:3),
2.88 ± 0.04 µm (CGF1:4) as the ratio of fucoidan increase continuously.
The trend was consistence with the value of d4,3. It is also believed that a
good emulsifier tended to provide a small droplet size. However, a
similar phenomenon has been found in the high internal phase emulsion
gels stabilized by insoluble soy polysaccharide nanoparticles (c = 0.25
wt%) (Yang et al., 2020b), which may be explained by the phenomenon
of bridging flocculation. Flocculation refers to the combination of two or
more oil droplets, but maintain their integrity. In general, flocculation is
detrimental to the stability of emulsion. However, moderate bridging
flocculation will cause an increase of emulsions viscosity, which is
beneficial to the stability of emulsions.
To confirm our hypothesis, the microstructure of oil droplets in
emulsions was observed by inverted fluorescence microscopy. The oil
phase was stained by Nile red. As depicted in Fig. 2A, O/W emulsions
were successfully prepared. Compared to the emulsion stabilized by CG
or CG-heated, the oil droplet size stabilized by glycated CGF complexes
decreased as the ratio of CG and fucoidan from 10:1 to 1:1. However, the
oil droplets occurred flocculation phenomenon when the CG and
fucoidan ratio was 1:2, 1:3, 1:4. This could explain why the droplet size
of oil phase increased. In addition, the binding activity between oil
droplets was strong to resist the stirring and shaking during the light
scattering measurements. The strong interaction between two or more
neighboring droplets may be attributed to the viscoelastic interfacial
layer on the oil–water interface, which can hinder the movement of oil
droplets and prevent the breakage of oil droplets (Niu et al., 2023).
3.2.2. Rheology properties of emulsions
The rheological properties of emulsions play an important role in
their stability and functionality for the application. The emulsions with
20% MCT oil were prepared to study the rheological properties of CG
and glycated CGF complexes. As can be seen in Fig. S1, the apparent
viscosity of emulsions with shear rate was measured. The viscosity
gradually decreased with the increasing shear rate in the range of 1–100
s− 1, the difference of viscosity between CGF and CG were significant in
high speed rate (10–100 s− 1) than that at low shear rate (1–10 s− 1). The
phenomenon of shear thinning demonstrated that all samples were non-
Newtonian fluid (Liu et al., 2023). Moreover, the viscosity of emulsions
stabilized by glycated CGF complexes was higher than CG and CG-
heated, which may be beneficial to the greater stability. With CGF1:4
Q. Zhu et al. Food Chemistry 430 (2024) 137033

Fig. 2. Microstructure images (A), Evolution of transmission profiles (B), Evolution of integral transmission (C), the change rates of the integral transmission (D),
evolution of instability index (E) and the instability index (F) of emulsions stabilized by 1% CG and glycated CGF complexes.

6
Q. Zhu et al. Food Chemistry 430 (2024) 137033

Fig. 3. Time dependence of interfacial tension (A), the square root of time dependence of interfacial pressure (π, mN/m) (B), time dependence of ln[(π10800-πt) /
(π10800-π0)] (C), interfacial viscoelastic modulus as a function of interfacial pressure (D), time evolution of interfacial elastic modulus (E) and time evolution of tanφ
(F), interfacial viscoelastic modulus as a function of amplitude (G) and frequency (H) of 1% CG and glycated CG complexes.

7
Q. Zhu et al.
having the highest viscosity, which was consistence with the macro
observation (data not shown). Chen et al. (2023) also verified that the
droplets movement was more easily limited by higher concentration of
Mesona chinensis Benth polysaccharide, and improved the stability of
emulsions.
3.2.3. Long-term stability analysis of emulsions
The emulsion was initially an opaque and uniform system. According
to the standard procedure, the emulsions were gradually centrifuged
using LUMiSizer to accelerate instability. The typical transmission pro­
file change of CG and glycated CGF complex with time is presented in
Fig. 2B. During the process of rapid centrifugation, the oil phase
migrated upward owing to low density, while the aqueous phase with
higher transparency remained at the bottom. As the area of transmission
profile decreased, the emulsion tended to be more stable. The first
transmission profile of emulsions prepared by natural CG and CG-heated
were far away from zero, which reflected that emulsions were extremely
unstable. Emulsions stabilized by glycated CGF complexes became sta­
ble with an increase in the ratio of fucoidan. In other words, the physical
stability of the emulsions was enhanced (Ma et al., 2023).
To facilitate the comparison of transmission changes, the integral
transmission profiles with time were plotted in Fig. 2C. The slopes of
transmission profiles were shown in Fig. 2D, which reflected the rates of
transmission. The value of slope is inversely proportional to the stability
of emulsion. The rate of integral transmission decreased with the in­
crease ratio of fucoidan to CG in glycated system, and emulsions became
more stable than the control. The change in the instability index over
time and instability index at the ending of separation were shown in
Fig. 2E and F. The range of the instability index is from 0 to 1, 0 means
that the sample is very opaque and stable, while 1 is extremely unstable
(Fernandes et al., 2017). Initially, the emulsion was relatively uniform,
which separated with time, so the clarification index increased. The
instability index of emulsion prepared by glycated CGF complexes
decreased compared to the control. It reflects that emulsion stabilized by
glycated CGF complexes was more stable. The instability index of
emulsion prepared by 1% CGF1:4 was 0.439 ± 0.029, which may be
owing to the higher viscosity and the thicker film on the interface. Ac­
cording to previous study, moderate bridging flocculation is beneficial
to the stability of emulsions owing to the higher viscosity (Li et al.,
2021).
3.3. Interfacial properties of CGF complexes on the oil–water interface
3.3.1. Dilatational interfacial tension and interfacial dilatational rheology
analysis
To better understand the behavior of CG and glycated CGF com­
plexes on oil–water interface, the interfacial tension was measured for 3
h. As shown in Fig. 3A, the interfacial tension of all samples decreased
gradually with the time due to the continuous and spontaneous
adsorption of CG and glycated CGF complexes on the surface of oil
droplets. After adsorption equilibrium for 3 h, the interfacial tension of
CG and CGF1:4 were 6.68 mN/m and 7.91 mN/m respectively. Glycated
CGF complexes showed higher interfacial tension than CG, indicating
the decrease in affinity on interface. A similar phenomenon was
demonstrated in soy protein isolate-gum Arabic composites stabilized
emulsion (Feng et al., 2023). This result suggested that reducing inter­
facial tension may not be the main reason for better emulsifying per­
formance of CGF complexes.
Fig. 3B displayed the curves of interfacial pressure (π) verus the
square root of time (t1/2) of CG and glycated CGF complexes. Compared
to CG, glycated CGF complexes had lower interfacial pressure, which
may be attributed to the larger hydrodynamic radii of glycated CGF
complexes. The characteristic limited the adsorption of protein on the
surface of oil droplets. Another study demonstrated that glycated whey
protein isolate and chitooligosaccharide exhibited lower initial π values
(Qin et al., 2022). In addition, slope of the curves is called diffusion rate
8
Food Chemistry 430 (2024) 137033
constant (Kdiff). The rate of adsorption slowed down after the short
disffusion-controlled adsorption process, which was attributed to the
presence of energy barriers (Niu et al., 2022a). Fig. 3C showed the
curves of ln[(π10800-πt)/(π10800-π0)] over time for CG and glycated CGF
complexes. The curve exhibited two linear regions generally, the initial
and second slope represented the penetration rate (Kp) and rearrange­
ment rate (Kr), respectively. The addition of fucoidan reduced the value
of Kdiff in solutions, Kp and Kr also exhibited the same tendency
(Table S1). It might be attributed to the molecular entanglement and
high viscosity in the system prolonged the diffusion, penetration and
rearrangement rate of CGF molecules.
As shown in Fig. 3D, the E-π curves were used to present the visco­
elasticity of interfacial layers on oil–water interface. E increased with
the increase in interfacial pressure, indicating the continuous adsorption
of CG and glycated CGF complexes at interface, forming viscoelastic
films. When the ratio of CG to fucoidan was 1:3 and 1:4, the E-π plots
shifted towards lower π values, which indicated an increased accumu­
lation of the CGF complex on the interface. In addition, the slopes of E-π
curves were greater than 1, which indicated strong molecular interac­
tion to form viscoelastic film (data not shown). Fig. 3E showed the
curves of elastic modulus over time. Ed gradually increased with
continuous adsorption of CG and glycated CGF complexes on the
interface, and reached equilibrium finally. Ed showed a slight growth in
later stages, indicating the molecules reorganization on the interface.
These results were also found in interfacial behavior of Mesona chinensis
Benth polysaccharide (Chen et al., 2023). Fig. 3F showed the change of
tanφ over time for CG and glycated CGF complexes. The tanφ value of all
samples is<1 (data not shown), indicating elasticity is the main property
of the interface (Bouyer et al., 2012). The smaller value of tanφ means
the better elastic properties.
3.3.2. Amplitude and frequency sweeps
Amplitude and frequency sweeps were adopted to evaluate me­
chanical properties of the interface layer. The change of viscoelastic
modulus with amplitude sweeps was shown in Fig. 3G. Viscoelastic
modulus of all samples gradually decreased with the amplitude defor­
mation from 5% to 30%, which indicated the interfacial layer was
damaged (Niu et al., 2022b). According to the principle of Fourier
transform that was used to calculate viscoelastic modulus, the nonline­
arity existed in original signal were ignored. Thus, the Lissajous plots
retain original data were used to illustrate the nonlinear dilatational
behavior (Fig. S2). As the deformation of ΔA/A increased, the curve of
Lissajous plots became wider, which was attributed to the response of
viscosity (Yang et al., 2020a). When the amplitude sweep (ΔA/A =
30%), the slope of all sample curves decreased during extension,
implying that the interfacial layers underwent strain softening. In
contrast, the slope of curves increased during compression deformation,
which suggested that the interfacial layer experienced strain stiffening.
Tang & Shen (2015) also showed BSA molecules exhibited the similar
phenomenon. Fig. 3H displayed the change of viscoelastic modulus with
frequency, E increased with frequency. This phenomenon can be
explained by diffusion relaxation. At low frequency of perturbation,
there were enough time for the CG and CGF complexes to the interface,
resulting a small viscoelastic modulus exhibited by the interfacial layer.
However, when subjected to high frequency perturbation, the short
detection time limited the diffusion relaxation of the interfacial layer,
showed a large viscoelastic modulus (Ma et al., 2023).
3.3.3. QCM-D analysis
QCM-D is an effective tool to study the adsorption processing of
molecules on the interface. According to Sauerbrey equation, there is a
linear relationship between the change of frequency and mass, so the
thickness of adsorption layer can be calculated. Moreover, dissipation
and energy loss are directly proportional, and can reflect the rheological
information about the adsorption layer. Based on macroscopic obser­
vation, we selected CG and four glycated CGF complexes that were
Q. Zhu et al. Food Chemistry 430 (2024) 137033

Fig. 4. Frequency change Δf (A) and dissipation change ΔD (B), ΔD-Δf curves of adsorption (C) and desorption (D) at n = 3 overtones, mass (E) and thickness (F) of
interfacial layer of glycated CGF complexes (1%).

unstable (CGF2:1) and relatively stable (CGF1:2, CGF1:3, CGF1:4) on
emulsions for QCM-D analysis. Fig. 4 shows the curve of Δf (A), ΔD (B),
ΔD-Δf of adsorption (C) and desorption (D), the change of mass (E) and
thickness (F) about 1% CG and glycated CGF complexes on the
adsorption layer (n = 3 overtones). Initially, the gold sensors coated
with MCT oil were balanced by buffer solution. With the evolution of
time, there was a continuous absorption and reorganization of CG and
CGF covalent complexes at the interface between oil and water, which
led to a gradual decrease in frequency and an increase in dissipation
(Fig. 4A, 4B). The dissipation increased rapidly with the molecules

9
Q. Zhu et al.
Fig. 5. Schematic mechanism of emulsions stabilized by CG and glycated CGF complexes.
adsorption on the interface continuously (Chen et al., 2023). The ΔD-Δf
curves of adsorption and desorption were shown in Fig. 4C and 4D, the
slope of K value indicates the energy loss with the change of per fre­
quency. In our results, the curves were continuous, which indicated that
glycated CGF complexes were a slow adsorption and desorption process
on the oil–water interface (Wei et al., 2020). In addition, the slopes of
ΔD-Δf curves were discontinuous, K2＜K1 (data not shown), indicated
that the conformation of glycated CGF complexes has been changed,
maybe owing to the multilayer adsorption of CGF covalent complexes on
the interface. And K3＞K4 at the desorption curve, the interfacial layer
became dense from loose. As reported before, whey protein also un­
dergone a conformational transition on the interface (Li et al., 2021).
The change of interfacial layer mass and thickness was consistent with
the Δf (Fig. 4E, F). The covalent complexes with high-ratio fucoidan had
obvious advantages in interfacial layer thickness, the layer thickness
was 32.05 ± 0.03 nm stabilized by 1% CGF1:4. The layer thickness of CG,
CGF2:1, CGF1:2, CGF1:3 were 15.87 ± 0.01 nm, 15.16 ± 0.03 nm, 20.23
± 0.02 nm, 29.33 ± 0.04 nm, respectively. This could explain why
glycated CGF complexes with the better emulsifying stability.
3.4. Schematic mechanism of glycated CGF complexes formation and
their behavior at o/w interface
The interaction of fucoidan with cocosin was simulated by molecular
docking method. As shown in Fig. 5, the aldehyde group of fucoidan
might be attached to the amino acid residue (Arg 196) on the surface of
cocosin, with a distance of 2.94 Å. The binding energy was − 5.42 kcal/
mol, indicating that it was easy to form glycated conjugates between
cocosin and fucoidan at these locations. In addition, as confirmed by the
interfacial dilatational rheology and QCM-D analysis, glycated CGF
complexes with a high ratio of fucoidan could form a thicker elastic film
at the O/W interface. In this regard, stronger electrostatic repulsions and
steric hindrance between droplets were beneficial to the better
10
Food Chemistry 430 (2024) 137033
emulsifying performance of glycated CGF complexes.
4. Conclusion
In conclusion, glycated coconut globulins-fucoidan (CGF) complexes
was fabricated to evaluate their emulsifying performance and interfacial
behavior. The results showed that glycated CGF complexes with high
grafting degree exhibited better emulsifying stability due to the higher
viscosity and a thicker elastic interfacial layer formed on the interface,
simultaneously provided higher electronegativity in emulsions. It was
suggested that appropriate glycosylation of CG with fucoidan could
improve the physicochemical characterize and long-term stability of
emulsions. These findings are useful for the application of glycated
protein-polysaccharide in food emulsions, and can serve as reference for
high-value utilization of coconut protein.
CRediT authorship contribution statement
Qianqian Zhu: Methodology, Investigation, Data curation, Writing –
original draft. Haiming Chen: Writing – review & editing, Funding
acquisition. Weijun Chen: Data curation. Ming Zhang: Methodology,
Data curation. Qiuping Zhong: Validation. Zixin Chen: Investigation.
Jianfei Pei: Software. Wenxue Chen: Conceptualization, Supervision.
Declaration of Competing Interest
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influence
the work reported in this paper.
Data availability
No data was used for the research described in the article.
Q. Zhu et al. Food Chemistry 430 (2024) 137033

Acknowledgements Liu, X., Zhang, X., Ding, L., Jin, H., Chen, N., Huang, X., Jin, Y., & Cai, Z. (2023). Natural
egg yolk emulsion as wall material to encapsulate DHA by two-stage
homogenization: Emulsion stability, rheology analysis and powder properties. Food
This work was supported by Hainan Province Science and Technol­ Research International, 167, Article 112658. https://doi.org/10.1016/j.
ogy Special Fund (ZDYF2022XDNY166) and Hainan Provincial Natural foodres.2023.112658
Science Foundation of China (223RC402). Lu, F., Ma, Y., Zang, J., Qing, M., Ma, Z., Chi, Y., & Chi, Y. (2023). High-temperature
glycosylation modifies the molecular structure of ovalbumin to improve the freeze-
thaw stability of its high internal phase emulsion. International Journal of Biological
Appendix A. Supplementary data Macromolecules, 233, Article 123560. https://doi.org/10.1016/j.
ijbiomac.2023.123560
Ma, J., Chen, H., Chen, W., Wu, J., Li, Z., Zhang, M., Zhong, Q., & Chen, W. (2022).
Supplementary data to this article can be found online at https://doi. Effects of heat treatment and pH on the physicochemical and emulsifying properties
org/10.1016/j.foodchem.2023.137033. of coconut (Cocos nucifera L.) globulins. Food Chemistry, 388, Article 133031.
https://doi.org/10.1016/j.foodchem.2022.133031
Ma, J., Pan, C., Chen, H., Chen, Y., Chen, W., Pei, J., … Chen, W. (2023). Interfacial
References: behavior of coconut (Cocos nucifera L.) globulins at different pH: Relation to
emulsion stability. Food Hydrocolloids, 144, Article 108958. https://doi.org/
10.1016/j.foodhyd.2023.108958
Ai, M., Xiao, N., & Jiang, A. (2021). Molecular structural modification of duck egg white
Martinez-Padilla, L. P., Hernandez-Rojas, F. S., Sosa-Herrera, M. G., & Juliano, P. (2022).
protein conjugates with monosaccharides for improving emulsifying capacity. Food
Novel application of ultrasound and microwave-assisted methods for aqueous
Hydrocolloids, 111, Article 106271. https://doi.org/10.1016/j.
extraction of coconut oil and proteins. Journal of Food Science and Technology, 59
foodhyd.2020.106271
(10), 3857–3866. https://doi.org/10.1007/s13197-022-05409-0
Alghazwi, M., Smid, S., Karpiniec, S., & Zhang, W. (2019). Comparative study on
Niu, H., Chen, X., Luo, T., Chen, H., & Fu, X. (2022a). Relationships between the
neuroprotective activities of fucoidans from Fucus vesiculosus and Undaria pinnatifida.
behavior of three different sources of pectin at the oil-water interface and the
International Journal of Biological Macromolecules, 122, 255–264. https://doi.org/
stability of the emulsion. Food Hydrocolloids, 128, Article 107566. https://doi.org/
10.1016/j.ijbiomac.2018.10.168
10.1016/j.foodhyd.2022.107566
Bouyer, E., Mekhloufi, G., Rosilio, V., Grossiord, J. L., & Agnely, F. (2012). Proteins,
Niu, H., Chen, X., Luo, T., Chen, H., & Fu, X. (2022b). The interfacial behavior and long-
polysaccharides, and their complexes used as stabilizers for emulsions: Alternatives
term stability of emulsions stabilized by gum arabic and sugar beet pectin.
to synthetic surfactants in the pharmaceutical field? International Journal of
Carbohydrate Polymers, 291, Article 119623. https://doi.org/10.1016/j.
Pharmaceutics, 436(1–2), 359–378. https://doi.org/10.1016/j.ijpharm.2012.06.052
carbpol.2022.119623
Chen, X., Niu, H., Chen, H., Zhu, X., Shen, M., & Xie, J. (2023). Interfacial behavior of
Niu, H., Wang, W., Dou, Z., Chen, X., Chen, X., Chen, H., & Fu, X. (2023). Multiscale
Mesona chinensis Benth polysaccharide at the oil-water interface and the stability of
combined techniques for evaluating emulsion stability: A critical review. Advances in
the emulsion. Food Hydrocolloids, 143, Article 108847. https://doi.org/10.1016/j.
Colloid and Interface Science, 311, Article 102813. https://doi.org/10.1016/j.
foodhyd.2023.108847
cis.2022.102813
Ding, Y., Chen, L., Shi, Y., Akhtar, M., Chen, J., & Ettelaie, R. (2021). Emulsifying and
Patil, U., & Benjakul, S. (2017). Characteristics of albumin and globulin from coconut
emulsion stabilizing properties of soy protein hydrolysates, covalently bonded to
meat and their role in emulsion stability without and with proteolysis. Food
polysaccharides: The impact of enzyme choice and the degree of hydrolysis. Food
Hydrocolloids, 69, 220–228. https://doi.org/10.1016/j.foodhyd.2017.02.006
Hydrocolloids, 113, Article 106519. https://doi.org/10.1016/j.
Patil, U., & Benjakul, S. (2018). Coconut milk and coconut oil: Their manufacture
foodhyd.2020.106519
associated with protein functionality. Journal of Food Science, 83(8), 2019–2027.
Drusch, S., Klost, M., & Kieserling, H. (2021). Current knowledge on the interfacial
https://doi.org/10.1111/1750-3841.14223
behaviour limits our understanding of plant protein functionality in emulsions.
Pirestani, S., Nasirpour, A., Keramat, J., Desobry, S., & Jasniewski, J. (2018). Structural
Current Opinion in Colloid & Interface Science, 56, Article 101503. https://doi.org/
properties of canola protein isolate-gum Arabic Maillard conjugate in an aqueous
10.1016/j.cocis.2021.101503
model system. Food Hydrocolloids, 79, 228–234. https://doi.org/10.1016/j.
EI-Hadary, A. R. E., Sulieman, A. M., & EI-Shorbagy, G. A. (2023). Comparative effects of
foodhyd.2018.01.001
hibiscus leaves and potato peel extracts on characteristics of fermented orange juice.
Tang, C.-H., & Shen, L. (2015). Dynamic adsorption and dilatational properties of BSA at
Journal of Food Quality and Hazards Control. 10, 39-50. https://doi.org/10.18502/
oil/water interface: Role of conformational flexibility. Food Hydrocolloids, 43,
jfqhc.10.1.11988.
388–399. https://doi.org/10.1016/j.foodhyd.2014.06.014
Fan, Y., Yi, J., Zhang, Y., Wen, Z., & Zhao, L. (2017). Physicochemical stability and in
Qin, X., Yu, J., Wang, Q., Zhang, H., Chen, H., Hu, Z., Lv, Q., & Liu, G. (2022).
vitro bioaccessibility of β-carotene nanoemulsions stabilized with whey protein-
Preparation of camellia oil Pickering emulsion stabilized by glycated whey protein
dextran conjugates. Food Hydrocolloids, 63, 256–264. https://doi.org/10.1016/j.
isolate and chitooligosaccharide: Effect on interfacial behavior and emulsion
foodhyd.2016.09.008
stability. LWT, 153, Article 112515. https://doi.org/10.1016/j.lwt.2021.112515
Feng, S., Guo, Y., Liu, F., Li, Z., Chen, K., Handa, A., & Zhang, Y. (2023). The impacts of
Raghavendra, A. N. S. N., & Raghavarao, K. S. M. S. (2012). Production of coconut
complexation and glycated conjugation on the performance of soy protein isolate-
protein powder from coconut wet processing waste and its characterization. Applied
gum Arabic composites at the o/w interface for emulsion-based delivery systems.
Biochemistry and Biotechnology, 167(5), 1290–1302. https://doi.org/10.1007/
Food Hydrocolloids, 135, Article 108168. https://doi.org/10.1016/j.
s12010-012-9632-9
ijbiomac.2023.123554
Su, J., Huang, Z., Yuan, X., Wang, X., & Li, M. (2010). Structure and properties of
Fernandes, A. R., Ferreira, N. R., Fangueiro, J. F., Santos, A. C., Veiga, F. J., Cabral, C.,
carboxymethyl cellulose/soy protein isolate blend edible films crosslinked by
Silva, A. M., & Souto, E. B. (2017). Ibuprofen nanocrystals developed by 22 factorial
Maillard reactions. Carbohydrate Polymers, 79(1), 145–153. https://doi.org/
design experiment: A new approach for poorly water-soluble drugs. Saudi
10.1016/j.carbpol.2009.07.035
Pharmaceutical Journal, 25(8), 1117–1124. https://doi.org/10.1016/j.
Thaiphanit, S., & Anprung, P. (2016). Physicochemical and emulsion properties of edible
jsps.2017.07.004
protein concentrate from coconut (Cocos nucifera L.) processing by-products and the
Gerliani, N., Hammami, R., & Aïder, M. (2019). Assessment of the extractability of
influence of heat treatment. Food Hydrocolloids, 52, 756–765. https://doi.org/
protein-carbohydrate concentrate from soybean meal under acidic and alkaline
10.1016/j.foodhyd.2015.08.017
conditions. Food Bioscience, 28, 116–124. https://doi.org/10.1016/j.
Wei, Y., Xie, Y., Cai, Z., Guo, Y., Wu, M., Wang, P., Li, R., & Zhang, H. (2020). Interfacial
fbio.2019.01.004
and emulsion characterisation of chemically modified polysaccharides through a
Lad, V. N., & Murthy, Z. V. P. (2012). Enhancing the stability of oil-in-water emulsions
multiscale approach. Journal of Colloid and Interface Science, 580, 480–492. https://
emulsified by coconut milk protein with the application of acoustic cavitation.
doi.org/10.1016/j.jcis.2020.07.048
Industrial & Engineering Chemistry Research, 51(11), 4222–4229. https://doi.org/
Xia, S., Li, Y., Xia, Q., Zhang, X., & Huang, Q. (2015). Glycosylation of bovine serum
10.1021/ie202764f
albumin via Maillard reaction prevents epigallocatechin-3-gallate-induced protein
Li, S., Sun, J., Yan, J., Zhang, S., Shi, C., McClements, D. J., Liu, X., & Liu, F. (2021).
aggregation. Food Hydrocolloids, 43, 228–235. https://doi.org/10.1016/j.
Development of antibacterial nanoemulsions incorporating thyme oil: Layer-by-layer
foodhyd.2014.05.022
self-assembly of whey protein isolate and chitosan hydrochloride. Food Chemistry,
Yang, H., Wang, S., Xu, Y., Wang, S., Yang, L., Song, H., He, Y., & Liu, H. (2023). Storage
339, Article 128016. https://doi.org/10.1016/j.foodchem.2020.128016
stability and interfacial rheology analysis of high-internal-phase emulsions stabilized
Li, Y., Liu, B., Jiang, L., Regenstein, J. M., Jiang, N., Poias, V., … Wang, Z. (2019).
by soy hull polysaccharide. Food Chemistry, 418, Article 135956. https://doi.org/
Interaction of soybean protein isolate and phosphatidylcholine in nanoemulsions: A
10.1016/j.foodchem.2023.135956
fluorescence analysis. Food Hydrocolloids, 87, 814–829. https://doi.org/10.1016/j.
Yang, J., Thielen, I., Berton-Carabin, C. C., van der Linden, E., & Sagis, L. M. C. (2020a).
foodhyd.2018.09.006
Nonlinear interfacial rheology and atomic force microscopy of air-water interfaces
Li, Z., Xi, J., Chen, H., Chen, W., Chen, W., Zhong, Q., & Zhang, M. (2022). Effect of
stabilized by whey protein beads and their constituents. Food Hydrocolloids, 101,
glycosylation with apple pectin, citrus pectin, mango pectin and sugar beet pectin on
Article 105466. https://doi.org/10.1016/j.foodhyd.2019.105466
the physicochemical, interfacial and emulsifying properties of coconut protein
Yang, T., Li, X.-T., & Tang, C.-H. (2020b). Novel edible pickering high-internal-phase-
isolate. Food Research International, 156, Article 111363. https://doi.org/10.1016/j.
emulsion gels efficiently stabilized by unique polysaccharide-protein hybrid
foodres.2022.111363
nanoparticles from Okara. Food Hydrocolloids, 98, Article 105285. https://doi.org/
Liu, Q., Chen, J., Qin, Y., Jiang, B., & Zhang, T. (2020). Zein/fucoidan-based composite
10.1016/j.foodhyd.2019.105285
nanoparticles for the encapsulation of pterostilbene: Preparation, characterization,
Zhang, Y., Tan, C., Abbas, S., Eric, K., Zhang, X., Xia, S., & Jia, C. (2014). The effect of soy
physicochemical stability, and formation mechanism. International Journal of
protein structural modification on emulsion properties and oxidative stability of fish
Biological Macromolecules, 158, 461–470. https://doi.org/10.1016/j.
ijbiomac.2020.04.128

11
Q. Zhu et al.
oil microcapsules. Colloids and surfaces. B, Biointerfaces, 120, 63–70. https://doi.org/
10.1016/j.colsurfb.2014.05.006
Zhang, Y., Zhang, Y., Chen, N., Xin, N., Li, Q., Ye, H., … Zhang, T. (2022). Glycated
modification of the protein from Rana chensinensis eggs by Millard reaction and its
stability analysis in curcumin encapsulated emulsion system. Food Chemistry, 382,
Article 132299. https://doi.org/10.1016/j.foodchem.2022.132299
12
Food Chemistry 430 (2024) 137033
Zhao, C., Chu, Z., Mao, Y., Xu, Y., Fei, P., Zhang, H., … Liu, J. (2023). Structural
characteristics and acid-induced emulsion gel properties of heated soy protein
isolate–soy oligosaccharide glycation conjugates. Food Hydrocolloids, 137, Article
108408. https://doi.org/10.1016/j.foodhyd.2022.108408