Food Hydrocolloids 95 (2019) 378–384

Contents lists available at ScienceDirect

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

The effects of amino acids on the gel properties of potassium iota T

carrageenan
Yingying Donga, Chengrong Wena,∗, Tingting Lia, Chunhua Wub, Hang Qia, Miao Liua,
Zhongli Wanga, Beiwei Zhua, Shuang Songa,∗∗
a
National Engineering Research Center of Seafood, National & Local Joint Engineering Laboratory for Marine Bioactive Polysaccharide Development and Application,
School of Food Science and Technology, Dalian Polytechnic University, Dalian, 116034, China
b
School of Food Science and Technology, Fujian Agriculture and Forestry University, Fuzhou, 116034, China

ARTICLE INFO ABSTRACT

Keywords: The effects of positive and neutral charged amino acids (Arginine (Arg) and Glutamine (Gln)) on the gel
Potassium iota carrageenan properties and microstructure of potassium iota Carrageenan (KIC) were investigated. The findings suggested
Glutamine that the gel melting temperature (Tg) of KIC gels increased from 36 °C to 43 °C with the Gln concentration
Arginine increasing, and the elastic modulus (G’) increased as well. However, with the increase Arg concentration, Tg and
Gel properties
G’ of gels increased gradually and then decreased, and the maximum Tg (46 °C) and G’ were obtained at con-
centrations of 0.05% Arg. LF-NMR results showed that the addition of Gln decreased the position of T23, in-
dicating a negative effect of Gln on the water holding capacity and water mobility of KIC gels, while the addition
of Arg increased the position of T23, showing the opposite effect. Compared to the pure KIC and Gln-KIC (GKIC),
Arg-KIC (AKIC) showed a more regular network structure and a stronger water holding capacity. In addition, few
chemical bonds were formed between amino acid and KIC. It is electrostatic attraction between KIC and Arg and
the crowding of Gln that contributed to better gel properties. Taken together, our results provide insights for the
further application of GKIC or AKIC gels in food and medical applications.

1. Introduction carrageenans. It has been known that KC and IC have very different
gelling behaviours, which depends on their structures, and are affected
Carrageenan is water-soluble linear sulfated polysaccharides ex- by cations, protein, polysaccharides and emulsifier through direct
tracted from red seaweeds of Rhodophyceae class (Prajapati, Maheriya, structural bridging, indirect structural bridging, helical interactions and
Jani, & Solanki, 2014), whose structure is based on a disaccharide units accumulation et al. (Liu, Zhan, Wan, Wang, & Wang, 2015; Rhein-
with alternating (1 → 3)-linked-β-D-galactose and (1 → 4)-linked-α-D- Knudsen, Ale, & Meyer, 2015). At high temperature, carrageenan mo-
galactose (Liu & Li, 2016; Paşcalău et al., 2012). It has been widely used lecules exist as random coil structures, and upon cooling, the polymer
as thickening, stabilizing and gelling agent in food (Campo, Kawano, Jr, chains change conformation to helical structure. Further cooling, KC
& Carvalho, 2009; Velde, Knutsen, Usov, Rollema, & Cerezo, 2002) and will appear spiral dimer aggregation and form stable three-dimensional
pharmaceutical industries (Janaswamy, Gill, Campanella, & Pinal, network, which can form strong and rigid gels. For IC, the helices are
2013; Li, Ni, Shao, & Mao, 2014). According to the position and number not (or only very weakly) stabilized by aggregation, which can form soft
of sulfate groups and non-dehydrated α-D-galactopyranose rings, car- gels (Brenner, Tuvikene, Parker, Matsukawa, & Nishinari, 2014; Nanna,
rageenan is divided into various types (Du, Brenner, Xie, & Matsukawa, Marcel, & Anne, 2015).
2016), while only kappa carrageenan (KC), lambda carrageenan (LC) IC is the most highly sulfated of the helix-forming polysaccharides,
and iota carrageenan (IC) are the most commercially exploited containing two sulfate groups per disaccharide repeat unit, being a

Abbreviations: IC, iota carrageenan; KC, kappa carrageenan; LC, lambda carrageenan; KIC, potassium iota Carrageenan; Gln, glutamine; Arg, arginine; GKIC,
glutamine and potassium iota Carrageenan combined gels; AKIC, arginine and potassium iota Carrageenan combined gels; LF-NMR, low field nuclear magnetic
resonance; DSC, differential scanning calorimetry; Cryo-SEM, cryogenic scanning electron microscopy; FT-IR, fourier transform infrared spectroscopy; Tg, the gel
melting temperature
∗
Corresponding author. School of Food Science and Technology, Dalian Polytechnic University, No. 1 Qinggongyuan, Ganjingzi district, Dalian, 116034, PR China.
∗∗
Corresponding author.
E-mail addresses: [email protected] (C. Wen), [email protected] (S. Song).

https://doi.org/10.1016/j.foodhyd.2019.04.023
Received 22 September 2018; Received in revised form 8 April 2019; Accepted 12 April 2019
Available online 13 April 2019
0268-005X/ © 2019 Published by Elsevier Ltd.
Y. Dong, et al. Food Hydrocolloids 95 (2019) 378–384

high-molecular weight linear polymer (Ataman & Pekcan, 2007). 1.152%–7.226%, 0.138%, and 0.73%, respectively, as determined by
However, higher levels of sulfate groups resulted in lower gel strength, atomic emission spectrometry.
thus the IC has weaker gel properties than that of KC (Janaswamy &
Chandrasekaran, 2008). Metal ions were contained in commercial IC 2.3. Gel preparation
and play an important role in accelerating gelation of IC by different
action mechanisms according to charge number (Robal et al., 2016). The gels of amino acid and KIC were prepared as follows: 100 mg of
The sodium IC formed double helices in a trigonal net and stabilized by KIC was dissolved in 10 mL of deionized water to prepare 1% (w/v) KIC
either SO3−-NaeSO3- or SO3−-Na-water-NaeSO3- interactions solution. Amount of amino acid was dispersed in KIC solution for
(Janaswamy & Chandrasekaran, 2001). The calcium IC formed a 12 h at room temperature and heated with magnetic stirring at 90 °C for
threefold, right-handed, half-staggered, parallel, double helix stabilized 30 min. Then the solution was kept at 4 °C before measurements. The
by inter-chain hydrogen bonds, in which Ca2+ not only connect and concentrations of amino acids were 0.02%, 0.05%, 0.1% and 0.2% (w/
balance the charge on two sulfate groups from different helices, but also v), respectively. The gels of Gln-KIC and Arg-KIC were labeled as
brought the helices closer (Janaswamy & Chandrasekaran, 2002). GKIC1, GKIC2, GKIC3, GKIC4, AKIC1, AKIC2, AKIC3 and AKIC4, re-
Furthermore, the potassium IC was also confirmed to form double helix spectively. KIC gel without amino acid was prepared in the same con-
dimers by using High-sensitivity differential scanning calorimetry dition and used as a control group. As the concentration of Gln and Arg
(Grinberg et al., 2001). The X-ray fiber diffraction pattern of potassium increased, the pH of KIC changed from 9.72 to 8.5 and 10.66, respec-
IC shows that it is a polycrystalline and well-oriented fiber, whose unit tively.
cell is 9 times as large as the sodium cell and the rubidium cell, and is
four times that of the calcium cell, which can contain up to 27 helices in 2.4. Rheological measurements
its trigonal unit cell (Janaswamy & Chandrasekaran, 2002,2005), so
potassium IC (KIC) is selected to prepare the gel. The rheological properties were evaluated using a DHR-1 Discovery
Amino acids are increasingly applied as nutrient supplements and Rheometer (TA-Instruments-Waters LLC, US). An aluminum parallel
additives in the food industry, which are always used together with plate crosshatched geometry of 40 mm diameter was used and the
carrageenan in food. Amino acids showed significant influences on the geometry gap was set at 1000 μm. Linear viscoelastic region was
gel properties of hydrocolloids and the addition of charge-carrying identified from strain sweep measurements in the range 0.01–100%
amino acids had stronger effect than the neutral one. However, there is strain at 1 Hz and 25 °C. The obtained linear viscoelastic region range
no systematic study about effects of charge-carrying or neutral amino was from 0.1% to 10%. Thereby the strain amplitude was carried out at
acids on gel properties of carrageenan. It has been reported that the 1% to ensure that all of the measurements were carried out within the
sulfate groups on polysaccharides could interact with positively linear viscoelastic range. Frequency sweep tests in the range of
charged sites available from protein moieties, such as ε-amino, α- 0.1–10 Hz were obtained at 25 °C. Temperature ramp tests were con-
amino, guanidinium, and imidazole (Palanisamy, Töpfl, Aganovic, & ducted in the range 5–60 °C at 1 Hz, and the rate of heating was 3 °C/
Berger, 2018). However, the interaction mechanism between carra- min. All experiments were performed at least in triplicate.
geenan and amino acids has rarely been reported. The Gln and Arg are
two typical amino acids that contain α-amino groups, whose pI points 2.5. LF-NMR spin-spin relaxation time (T2) measurements
are 5.65 and 10.76, respectively. As the increasing concentration of Gln
and Arg, the pH of KIC should be between 5.65 and 10.76, which affects Transverse relaxation T2 measurement was performed following the
the chargeability of Gln and Arg, so it may potentially influence the gel method (Bi et al., 2016) with some modification. In brief, the tem-
properties of carrageenan. Thus, the objective of this work is to in- perature of the LF-NMR analyzer (Shanghai Niumag Corporation,
vestigate the effects of Gln and Arg on the gel properties and micro- Shanghai China) was set and maintained at 32 °C during the spin-spin
structure of potassium iota Carrageenan (KIC). The findings should relaxation time (T2) measurements, which were performed using a se-
provide some useful information for the design of KIC-amino acid quence based on the Carr-Purcell-Meiboom-Gill (CPMG) sequence. The
complex to achieve desired rheological, gelation and thermal properties instrument equipped with a 0.5 T permanent magnet corresponding to a
for food application. proton resonance frequency of 21 MHz. Data from 10000 echoes were
acquired as 4 scan repetitions, and pulse widths at 90° (P1) and 180°
2. Materials and methods (P2) of 27 and 54 μs. The repetition time between subsequent scans was
0.5s, and each measurement was performed in triplicate.
2.1. Materials
2.6. Differential scanning calorimetry (DSC) measurements
Iota carrageenan was purchased from Aladdin Co. Ltd. (Shanghai,
China). Glutamine and Arginine were purchased from Bioengineering The thermal properties of freeze-dried solution samples were mea-
Co. Ltd. (Shanghai, China). Isopropyl alcohol and KCl were purchased sured according to the method (Zhao et al., 2015) with some mod-
from Tianjin Damao chemical reagent plant (Tianjin, China). All che- ification by a calorimeter (DSC250, TA Instruments, USA). Briefly, 8 mg
micals were analytical grade. sample was put into an aluminum pan, and heated from 30 to 300 °C at
a heating rate of 5 °C/min. At the same time, an empty pan was per-
2.2. Potassium iota carrageenan preparation formed as control.

The KIC was prepared by the method (Polowsky & Janaswamy, 2.7. Cryo-SEM
2015) with some modification. 500 mg of IC was dissolved in 500 mL of
deionized water under constant magnetic stirring and heated to 95 °C The micro-structure of the gels was elucidated with cryogenic
and held for 60 min. Then 3 g of KCl was added to the hot solution with scanning electron microscopy (Cryo-SEM). The solution samples were
constant magnetic stirring at 95 °C for 60 min. The hot solution was placed in the slots of a stub, then placed at 4 °C for 10 min to form gels
poured into 1000 mL of pre-colling isopropyl alcohol (0 °C) to collect and then plunge-frozen in liquid nitrogen slush. Then the samples were
the white fibrous material. Later, the white fibrous material was rinsed transferred into the Cryo-preparation chamber (PP3010T Cryo-SEM
thoroughly in 80% and 100% isopropyl alcohol to remove unbound preparation system, Quorum Technologies, UK) with a treatment of
cations, and freeze dried for further study. The K+, Na+, and Ca2+ freeze-fractured, sublimated at −60 °C for 15min and sputtered with
contents of IC were changed from 4.05%, 1.896%, and gold at 10 mA for 40 s, examined with a SU8010 SEM (Hitachi Ltd.,

379
Y. Dong, et al. Food Hydrocolloids 95 (2019) 378–384

Japan) at an accelerating voltage of 12 kV. unstable AKIC gel network structure and easily destroyed. In short, Arg
could enhance KIC gel formation and the best gel stability was at 0.05%
2.8. FT-IR spectroscopy Arg concentration.
The G’ and tanδ (G''/G’) of GKIC and AKIC in the temperature range
Freeze-drying gel (1 mg) was mixed with 100 mg of dry KBr. The from 5 to 60 °C at 1 Hz were showed in Fig. 2. At low temperatures,
mixture was pressed into a disk for spectrum recording. The FTIR GKIC could maintain its structure, showing high G’ values (Fig. 2a).
spectra was recorded in the frequency range of 450–4000 cm−1 on a However, thermal motion of molecules in GKIC system were intensified
Frontier FTIR Spectrometer (Perkin Elmer, Waltham, USA), and the with the temperature increasing, which enlarged the intermolecular
resolution was set as 4 cm−1. distance and weakened the interactions (eg, hydrogen, electrostatic and
hydrophobic bonds), resulting in a slow decrease in the G’ values. It is
2.9. Statistical analyses known that tanδ is an important evaluation criterion for describing the
sol-gel transition. The temperature of tan (δ) = 1 was the gel melting
All measurements were carried out in triplicate, and spectra draw- temperature (Tg) (Torres, Chenlo, & Moreira, 2017). The Tg of KIC,
ings were performed using OriginPro 8.5. Significant differences be- GKIC1, GKIC2, GKIC3, and GKIC4 were around 36, 38, 39, 41 and
tween means (p < 0.05) were studied by one way ANOVA test using 43 °C, respectively, which increased with the concentration of Gln in-
the SPSS 22.0 statistical analysis program (SPSS Inc., Chicago, IL). creasing (Fig. 2b). The results suggested that Gln could enhance the
stability of the gel. As shown in Fig. 2c, the G’ of AKIC also increased
3. Results and discussion first and then decreased with the increase of Arg concentration like
Fig. 1c. The Tg of AKIC1, AKIC2, AKIC3 and AKIC4 were around 45, 47,
3.1. Rheological properties 44 and 43 °C, respectively (Fig. 2d). The maximum G’ and Tg of AKIC
were achieved at the concentration of 0.05% Arg. Moreover, G’ rapidly
The frequency dependence of GKIC and AKIC on elastic modulus decreased with the increase of temperature, indicated the weak inter-
(G’) and viscosity modulus (G″) in the frequency range of 0.1–10 Hz action between Arg and KIC.
were showed in Fig. 1. The concentration of Gln had an obvious effect Compared GKIC with AKIC, the G’ values of AKIC were higher than
on the elastic modulus (G’) (Fig. 1a) and viscosity modulus (G″) that of GKIC both in the frequency dependence and temperature de-
(Fig. 1b) of KIC, and the higher concentration of the Gln, the greater G’ pendence. In addition, the Tg of AKIC2 was higher than that of GKIC4.
and G″ were obtained. Moreover, G″ showed a weak dependence on However, there was a difference in the change rate of G’ with frequency
frequency and was always lower than G’ with the different concentra- and temperature, which was probably attributed to the different in-
tions of Gln, confirming a gel-like behavior of GKIC. It suggested that teraction between Gln and Arg with KIC. It has been reported that the
Gln could promote the formation of KIC gels with a concentration de- formation of mixed gel networks was related to many factors such as
pendence. In addition, G’ increased slowly with the increase of fre- electrostatic complexation, hydrogen bonding, hydrophobic interaction
quency, indicating a low frequency dependence and good gel stability (Antonov, Zhuravleva, Cardinaels, & Moldenaers, 2018). The formation
of GKIC gels. of K+ bridges in the KIC molecule exposed a large amount of negative
As shown in Fig. 1c and d, the G’ and G” values increased with the charge. Therefore, the rheological result of AKIC was probably due to
addition of Arg first, achieving a maximum curve at the Arg con- the electrostatic interaction between the positive charges of Arg and the
centration of 0.05%, and then decreased. G″ was always lower than G’ negative charge sulfate groups of KIC at low concentration of Arg.
with the different concentrations of Arg, which suggested a gel-like When the concentration of Arg was overmuch, the positive charge
behavior of AKIC. Compared with GKIC (Fig. 1a), G’ of AKIC showed a would excessive accumulation, which disrupted the structure of AKIC
large increase rate with the increase of frequency, indicating that an gels and formed discontinuous internal structures, resulting in a

Fig. 1. Effect of frequency on the elastic modulus (G’) and viscosity modulus (G″) of the GKIC (a and b) and AKIC (c and d) gels at 1% strain and 25 °C.

380
Y. Dong, et al. Food Hydrocolloids 95 (2019) 378–384

Fig. 2. Effect of temperature on the elastic modulus G’ and tanδ of the GKIC (a and b) and AKIC (c and d) gels at 1% strain and 1 Hz.

tendency for G’ and Tg to increase first and then decrease. However, Gln with macromolecules, the immobilized water and free water, respec-
is a neutral charged amino acid with negative charges in the GKIC gel tively (Zhang, Yang, Tang, Chen, & Yuan, 2015). GKIC and AKIC gel
system, which could have indirect interactions with the anionic carra- systems both were dominated by free water. Compared with KIC, the
geenan via trace divalent ions to promote the formation of KIC gel. addition of Gln induced an addition in the position of T23 and corre-
While due to the relatively low content of divalent ions in KIC system, sponding proportions (A23) (p < 0.05), which showed that the water
Gln may dominantly crowd in the gel network structure, which may be holding capacity of the gel system decreased with Gln concentration
the main cause of the increase in G’ and Tg of GKIC with the Gln increasing. This was probably due to the crowding of Gln in the GKIC
concentration increased. gel network, which weakened the water-holding capacity of GKIC gel.
In the AKIC gel system, Arg induced a reduction in the position and an
addition in A23 compared with the KIC (p < 0.05). Since the electro-
3.2. Low-field nuclear magnetic resonance (LF-NMR) relaxation
static attraction between Arg and KIC enhanced the formation of the gel
network, the water-holding capacity of the AKIC gel was improved. At
LF-NMR is a practical way to analyze proton relaxation in food
the same time, the water content of the gel network was increased
systems because of its non-invasive characteristics, high reproduci-
because of the water absorption of Arg. Compared to Gln, Arg had a
bility, and sensitivity (Bi et al., 2016; Geng et al., 2015). T2, the spin-
strong effect on the water-holding capacity of KIC gels.
spin relaxation time, provides information about the relaxation and
migration of hydrogen molecules, and is often regarded as an important
index estimating the mobility and distribution of different fractions of 3.3. DSC analysis
water molecules in the gel system (Ozel, Uguz, Kilercioglu, Grunin, &
Oztop, 2017; Pan, Guo, Li, Song, & Ren, 2017). T2 and A2 (the area The effects of Gln and Arg on the thermal properties of KIC were
proportion of each peak) of GKIC and AKIC gels are listed in Table 1. examined by DSC in the temperature range from 30 to 300 °C. The DSC
Three water fractions were distinguished in GKIC and AKIC gels. T21, curves of GKIC and AKIC were shown in Fig. 3. The exothermic peaks
T22 and T23 were identified to represent the water closely associated and endothermic peaks were observed on the DSC curves of GKIC

Table 1
The LF-NMR relaxation spectrum parameters (T21, T22, T23, A21, A22, and A23) of KIC as affected by Gln and Arg concentration.
Groups Time of relaxation (ms) The normalized peak area

T21 T22 T23 A21 A22 A23

ab a a a a
KIC 3.29 ± 0.89 39.53 ± 3.26 613.59 2.87 ± 0.64 7.18 ± 0.30 631.53 ± 2.79a
GKIC1 3.60 ± 1.00ab 52.25 ± 4.30b 705.48b 2.24 ± 1.16a 9.18 ± 0.60b 641.83 ± 1.90b
GKIC2 3.53 ± 0.49ab 60.08 ± 4.95bc 811.13c 2.60 ± 0.61a 9.91 ± 0.25c 641.76 ± 1.60b
GKIC3 4.94 ± 1.08a 83.20 ± 6.54d 932.60d 2.64 ± 0.28a 10.37 ± 0.27c 626.24 ± 1.38c
GKIC4 2.19 ± 0.78b 69.08 ± 5.69c 1072.27e 2.71 ± 0.60a 6.81 ± 0.30a 659.70 ± 2.37d
KIC 3.29 ± 0.89a 39.53 ± 3.26a 613.59a 2.87 ± 0.64a 7.18 ± 0.3a 631.53 ± 2.79a
AKIC1 2.95 ± 1.15a 27.43 ± 4.60b 533.67b 0.94 ± 0.65a 7.35 ± 0.33a 708.37 ± 3.48b
AKIC2 2.22 ± 1.20a 27.24 ± 2.14b 533.67b 1.82 ± 0.25a 6.35 ± 0.31b 682.60 ± 1.40c
AKIC3 1.66 ± 0.59a 15.59 ± 1.23c 464.16c 1.27 ± 0.24a 5.71 ± 0.38c 674.02 ± 8.36c
AKIC4 1.09 ± 0.58a 14.18c 403.70d 3.72 ± 2.85a 4.94 ± 0.37d 672.07 ± 12.95c

a-e
Means with different letters in the same column are significantly different (p＜0.05).

381
Y. Dong, et al. Food Hydrocolloids 95 (2019) 378–384

Fig. 3. The DSC curves of GKIC (a) and AKIC (b).

(Fig. 3a) and AKIC (Fig. 3b). The endothermic peak corresponded to the
evaporation of the sample powder (free water). The exothermic peak
indicated thermal degradation owing to the random breakage of the
glyosidic bonds and material devolatilization with evolution of the
volatile matter. And the higher the exothermic peak temperature, the
more stable thermal property of the material (Xu, Xue, Chang, Wang, &
Jiang, 2016). The endothermic peaks of GKIC and AKIC were very
small, indicating low water content in the samples. The exothermic
peaks of GKIC increased with the addition of Gln first, and then de-
creased, which were started at 216.7, 237.4, 235.6, 222.3 and 214.8 °C,
respectively, and the maximum temperature were 225.6, 245.3, 245.0,
231.9 and 222.1 °C for KIC, GKIC1, GKIC2, GKIC3 and GKIC4, respec-
tively. These results were probably owning to the crowding of Gln in
the GKIC gel network, which weakened the interaction between Gln
and KIC with the concentration of Gln increasing, resulting the thermal
properties first increased and then decreased. While the exothermic
Fig. 4. Cryo-SEM images of KIC, GKIC4, and AKIC4.
peaks were started at 247.0, 251.40, 254.0 and 256.4 °C, respectively,
and observed at the maximum temperature of 255.1, 259.0, 263.1 and
265.9 °C for AKIC1, AKIC2, AKIC3 and AKIC4, respectively. The in-
crease in the maximum temperature of exothermic peaks was due to
electrostatic interaction between KIC and Arg, resulting in an increase
in the thermal property of the gel.

3.4. Cryo-SEM

Electron microscopy is generally used as a technique for observing

the microstructure directly of the dry samples, which may change the
structure and original morphology of the samples. The cryogenic elec-
tron microscope can directly show the microstructure of aqueous
samples such as proteins and cells by rapidly freezing water-containing
samples and observing them in a frozen state at cryogenic temperatures
(Shimanuki et al., 2017). The microstructure of the gels observed by
Cryo-SEM was shown in Fig. 4. The three-dimensional network-like
microstructure of KIC was promoted by adding Gln and Arg. There were
large pores in the network structure of GKIC4, and the network struc-
tures were filled with many filamentous structures of Gln, which proved
that Gln crowded in the KIC gel network. Compared to the micro- Fig. 5. FT-IR of freeze-dried powders of KIC, GKIC, and AKIC gels.
structure of GKIC4, the microstructure of AKIC4 was denser and more
uniform. The pores of AKIC4 contained a small amount of filamentous
3.5. FT-IR spectroscopy
substances, suggesting that electrostatic interaction was generated be-
tween Arg and KIC. And the wall of the AKIC pore was more complete
Chemical interaction of KIC with Gln and Arg were evaluated using
and thicker than that of GKIC. The integrity of the network structure is
FT-IR spectroscopy, and the results were shown in Fig. 5. The broad
conducive to the enhancement of the water-holding capacity of gels.
band from 3200 to 3500 cm−1 was attributed to the stretching of inter-
Therefore, as well as LF-NMR, the Cryo-SEM results indicated that both
and intra-molecular bound hydroxyl groups (Paula et al., 2015). The
the gel formation ability and water-holding capacity of AKIC were
prominent peak around 2800-3000 cm−1 was corresponding to the
stronger than GKIC.
stretching vibration of CeH bonds (Martins et al., 2012). The char-
acteristic absorption peaks of hydroxyl group of KIC, GKIC2, GKIC4,
AKIC2 and AKIC4 were at 3431.51 cm−1, 3428.07 cm−1,

382
Y. Dong, et al. Food Hydrocolloids 95 (2019) 378–384

Fig. 6. The schematic mechanism for the effect of Gln (a) and Arg (b) on the gelation of KIC.

3429.01 cm−1, 3432.56 cm−1, and 3426.23 cm−1, respectively. The 4. Conclusion
addition of Gln and Arg caused the slight blue shift of the hydroxyl
absorption peak, which could prove that there was a weak hydrogen The present study has provided important insights in the gel prop-
bond interaction between Gln, Arg and KIC (Kong et al., 2014). The erties and microstructure of Gln-KIC and Arg-KIC mixed gels. The ad-
peak of KIC, GKIC2, GKIC4, AKIC2 and AKIC4 at 1636.66 cm−1, dition of Gln could induce KIC to form gels with a concentration de-
1631.87 cm−1, 1627.39 cm−1, 1630.85 cm−1, and 1629.27 cm−1 were pendence, but decreased the position of T23 and water holding capacity
both attributed to the structural water deformation band (Jumaah, in the gel system. Cryo-SEM imaging showed that the KIC gel network
Mobaraka, Ahmad, & Ramli, 2013). The presence of amide I in the structure changed from random to regular by adding Gln. The Gln was
range of 1700–1600 cm−1 indicated the electrostatic interaction be- negatively charged in the KIC solution, and the crowding of Gln played
tween the NH2 groups and the KIC sulfate groups (Du et al., 2013). a dominant role in improving KIC gel properties. However, the Arg was
There were no formation of other new functional groups, which in- positively charged in the KIC solution, which had different effect on the
dicated that there were no formation of new chemical bonds between properties of KIC gel. The addition of Arg increased water holding ca-
KIC with Gln and Arg. FTIR results indicated that there was a weak pacity and thermal stability with an optimal concentration, and the
hydrogen bonding interactions between Gln and Arg with KIC, and optimum rheological properties of AKIC were observed at a con-
confirmed the correctness of the hypothesis: Gln mainly crowded in the centration of 0.05% Arg. Compare to pure KIC and GKIC gels, AKIC gel
KIC gel network and there was electrostatic interaction between Arg had a more regular network structure. The different results suggested
and KIC. that the weak electrostatic interaction between Arg and KIC was the
primary factor to improve the properties of KIC gels. There is little re-
search on the gel properties of amino acid and KIC blends. Although the
3.6. The schematic mechanism improvement of KIC gel properties by adding Gln and Arg has been
observed in a limited number of tests, further studies must be per-
On the basis of the above experimental results, we propose a sche- formed before conclusions can be drawn on the effect of amino acid on
matic mechanism for the effect of Gln and Arg on the gelation of KIC as cation form carrageenan. And the blend gels could be applied as a new
shown in Fig. 6. It is well known that K+ can form inter- or in- find into the food, pharmaceutical and medicinal fields.
tramolecular cation-bridges with some of the sulfate groups of the IC,
which can connect the molecular chains of the IC to induce the for- Acknowledgements
mation of helices and space network structure. K+ ions acted as the
junctions of the gel network to promote the formation of KIC helixes, This work was financially supported by The National Natural
resulting in a large amount of negative charges exposed to the outside Science Foundation of China (No. 31501439). The authors are also
of KIC helixes. As a neutral charged amino acid, Gln mainly crowded in grateful to Dr. Haitao Wang for the technical assistance of Cryo-SEM
the gel network to improve the gel properties of KIC, rather than gen- measurements experiment.
erated electrostatic interaction with KIC. The positive charged Arg at-
tracted electrostatically with KIC, which promoted and intensified the References
formation of the gel network, but the excessive concentration of argi-
nine would cause charge repulsion, weakening the interaction between Antonov, Y. A., Zhuravleva, I. L., Cardinaels, R., & Moldenaers, P. (2018).
Arg and KIC, resulting in partial Arg dispersion in the gel network Macromolecular complexes of lysozyme with kappa carrageenan. Food Hydrocolloids,
74, 227–238.
structure. It is known that IC has a distinctive soft gel profiles due to the Ataman, E., & Pekcan, Ö. (2007). Small molecule sorption and desorption in and out of
differences in sulfate position, their proportion, and the presence of iota‐carrageenan gels. Journal of Macromolecular Science Part B, 46(4), 705–715.
anhydro-bridges. Many materials have been reported to promote IC gel Bi, J., Li, Y., Cheng, S., Dong, X., Kamal, T., Zhou, D., et al. (2016). Changes in body wall
of sea cucumber ( Stichopus japonicus ) during a two-step heating process assessed by
properties, such as divalent metal ions (Kara, Arda, & Pekcan, 2007), rheology, LF-NMR, and texture profile Analysis. Food Biophysics, 11(3), 257–265.
KC (Brenner et al., 2014), whey protein (Lam & Nickerson, 2014), Brenner, T., Tuvikene, R., Parker, A., Matsukawa, S., & Nishinari, K. (2014). Rheology
starch (Tischer, Noseda, Freitas, Sierakowski, & Duarte, 2006) et al. In and structure of mixed kappa-carrageenan/iota-carrageenan gels. Food Hydrocolloids,
39, 272–279.
this study, a monovalent cation form of IC was prepared, and the new
Campo, V. L., Kawano, D. F., Jr, D. B. D. S., & Carvalho, I. (2009). Carrageenans:
gels were formed by blending with amino acids. The new gels will Biological properties, chemical modifications and structural analysis - a review.
provide a new potential for IC applications in the food industry. Carbohydrate Polymers, 77(2), 167–180.
Du, L., Brenner, T., Xie, J., & Matsukawa, S. (2016). A study on phase separation behavior
in kappa/iota carrageenan mixtures by micro DSC, rheological measurements and
simulating water and cations migration between phases. Food Hydrocolloids, 55,

383
Y. Dong, et al. Food Hydrocolloids 95 (2019) 378–384

81–88. Ozel, B., Uguz, S. S., Kilercioglu, M., Grunin, L., & Oztop, M. H. (2017). Effect of different
Du, Y., Shi, S., Jiang, Y., Xiong, H., Woo, M. W., Zhao, Q., et al. (2013). Physicochemical polysaccharides on swelling of composite whey protein hydrogels: A low field (LF)
properties and emulsion stabilization of rice dreg glutelin conjugated with κ-carra- NMR relaxometry study. Journal of Food Process Engineering, 40(3), e12465. https://
geenan through Maillard reaction. Journal of the Science of Food and Agriculture, 93(1), doi.org/10.1111/jfpe.12465.
125–133. Palanisamy, M., Töpfl, S., Aganovic, K., & Berger, R. G. (2018). Influence of iota carra-
Geng, S., Wang, H., Wang, X., Ma, X. J., Xiao, S., Wang, J., et al. (2015). A non-invasive geenan addition on the properties of soya protein meat analogues. LWT- Food Science
NMR and MRI method to analyze the rehydration of dried sea cucumber. Analytical and Technology, 87, 546–552.
Methods, 7(6), 2413–2419. Pan, T., Guo, H., Li, Y., Song, J., & Ren, F. (2017). The effects of calcium chloride on the
Grinberg, V. Y., Grinberg, N. V., Usov, A. I., Shusharina, N. P., Khokhlov, A. R., & Kruif, K. gel properties of porcine myosin–κ-carrageenan mixtures. Food Hydrocolloids, 63,
G. d. (2001). Thermodynamics of conformational ordering of ι-carrageenan in KCl 467–477.
solutions using high-sensitivity differential scanning calorimetry. Biomacromolecules, Paşcalău, V., Popescu, V., Popescu, G. L., Dudescu, M. C., Borodi, G., Dinescu, A., et al.
2(3), 864–873. (2012). The alginate/k-carrageenan ratio's influence on the properties of the cross-
Janaswamy, S., & Chandrasekaran, R. (2001). Three-dimensional structure of the sodium linked composite films. Journal of Alloys and Compounds, 536, 418–423.
salt of iota-carrageenan. Carbohydrate Research, 335(3), 181–194. Paula, G. A., Benevides, N. M. B., Cunha, A. P., Oliveira, A. V. D., Pinto, A. M. B., Morais,
Janaswamy, S., & Chandrasekaran, R. (2002). Effect of calcium ions on the organization J. P. S., et al. (2015). Development and characterization of edible films from mixtures
of iota-carrageenan helices: An X-ray investigation. Carbohydrate Research, 337(6), ofκ-carrageenan, ι-carrageenan, and alginate. Food Hydrocolloids, 47, 140–145.
523–535. Polowsky, P. J., & Janaswamy, S. (2015). Hydrocolloid-based nutraceutical delivery
Janaswamy, S., & Chandrasekaran, R. (2005). Cation-induced polymorphism in iota- systems: Effect of counter-ions on the encapsulation and release. Food Hydrocolloids,
carrageenan. Carbohydrate Polymers, 60(4), 499–505. 43(43), 658–663.
Janaswamy, S., & Chandrasekaran, R. (2008). Heterogeneity in iota-carrageenan mole- Prajapati, V. D., Maheriya, P. M., Jani, G. K., & Solanki, H. K. (2014). Carrageenan: A
cular structure: Insights for polymorph II - > III transition in the presence of calcium natural seaweed polysaccharide and its applications. Carbohydrate Polymers, 105(1),
ions. Carbohydrate Research, 343(2), 364–373. 97–112.
Janaswamy, S., Gill, K. L., Campanella, O. H., & Pinal, R. (2013). Organized poly- Rhein-Knudsen, N., Ale, M. T., & Meyer, A. S. (2015). Seaweed hydrocolloid production:
saccharide fibers as stable drug carriers. Carbohydrate Polymers, 94(1), 209–215. An update on enzyme assisted extraction and modification technologies. Marine
Jumaah, F. N., Mobaraka, N. N., Ahmad, A., & Ramli, N. (2013). Characterization of ɽ Drugs, 13(6), 3340–3359.
-carrageenan and its derivative based green polymer electrolytes. Ukm fst postgraduate Robal, M., Brenner, T., Matsukawa, S., Ogawa, H., Truus, K., Rudolph, B., et al. (2016).
colloquium (pp. 768–774). . Monocationic salts of carrageenans: Preparation and physico-chemical properties.
Kara, S., Arda, E., & Pekcan, Ö. (2007). Monovalent and divalent cation effects on phase Food Hydrocolloids, 63, 656–667.
transitions of ι-carrageenan. Journal of Bioactive and Compatible Polymers, 22(1), Shimanuki, J., Takahashi, S., Tohma, H., Ohma, A., Ishihara, A., Ito, Y., et al. (2017).
42–61. Microstructural observation of fuel cell catalyst inks by Cryo-SEM and Cryo-TEM.
Kong, H., Yang, J., Zhang, Y., Fang, Y., Nishinari, K., & Phillips, G. O. (2014). Synthesis Microscopy, 66(3), 204–208.
and antioxidant properties of gum Arabic-stabilized selenium nanoparticles. Tischer, P. C. S. F., Noseda, M. D., Freitas, R. A. D., Sierakowski, M. R., & Duarte, M. E. R.
International Journal of Biological Macromolecules, 65, 155–162. (2006). Effects of iota-carrageenan on the rheological properties of starches.
Lam, R. S. H., & Nickerson, M. T. (2014). The properties of whey protein-carrageenan Carbohydrate Polymers, 65(1), 49–57.
mixtures during the formation of electrostatic coupled biopolymer and emulsion gels. Torres, M. D., Chenlo, F., & Moreira, R. (2017). Thermal reversibility of kappa/iota-hy-
Food Research International, 66(66), 140–149. brid carrageenan gels extracted from Mastocarpus stellatus at different ionic
Li, L., Ni, R., Shao, Y., & Mao, S. (2014). Carrageenan and its applications in drug de- strengths. Journal of the Taiwan Institute of Chemical Engineers, 71, 414–420.
livery. Carbohydrate Polymers, 103(1), 1–11. Velde, F. V. D., Knutsen, S. H., Usov, A. I., Rollema, H. S., & Cerezo, A. S. (2002). 1 H and
Liu, S., & Li, L. (2016). Thermoreversible gelation and scaling behavior of Ca2+ -induced 13 C high resolution NMR spectroscopy of carrageenans: Application in research and
κ-carrageenan hydrogels. Food Hydrocolloids, 61, 793–800. industry. Trends in Food Science & Technology, 13(3), 73–92.
Liu, J., Zhan, X., Wan, J., Wang, Y., & Wang, C. (2015). Review for carrageenan-based Xu, X., Xue, C., Chang, Y., Wang, J., & Jiang, K. (2016). Chain conformational and
pharmaceutical biomaterials: Favourable physical features versus adverse biological physicochemical properties of fucoidans from sea cucumber. Carbohydrate Polymers,
effects. Carbohydrate Polymers, 121, 27–36. 152, 433–440.
Martins, J. T., Cerqueira, M. A., Bourbon, A. I., Pinheiro, A. C., Bws, S., & Vicente, A. A. Zhang, Z., Yang, Y., Tang, X., Chen, Y., & Yuan, Y. (2015). Chemical forces and water
(2012). Synergistic effects between κ-carrageenan and locust bean gum on physico- holding capacity study of heat-induced myofibrillar protein gel as affected by high
chemical properties of edible films made thereof. Food Hydrocolloids, 29(2), 280–289. pressure. Food Chemistry, 188, 111–118.
Nanna, R. K., Marcel, A., & Anne, M. (2015). Seaweed hydrocolloid production: An up- Zhao, H., Zhou, F., Peng, W., Zheng, J., Dziugan, P., & Zhang, B. (2015). The effects of
date on enzyme assisted extraction and modification technologies. Marine Drugs, carrageenan on stability of arachin and the interactions between them. Food
13(6), 3340–3359. Hydrocolloids, 43, 763–768.

384