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Effects of wheat gliadin and glutenin fractions on dough properties, oil uptake
and microstructure of instant noodles

Article in Journal of Food Processing and Preservation · August 2022

DOI: 10.1111/jfpp.17100

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DOI: 10.1111/jfpp.17100

ORIGINAL ARTICLE

Effects of wheat gliadin and glutenin fractions on dough

properties, oil uptake, and microstructure of instant noodles

Aastha Dewan1 | Nisha Chaudhary2 | Bhupendar Singh Khatkar1

1
Department of Food Technology, Guru
Jambheshwar University of Science and Abstract
Technology, Hisar, India
In lieu to understand the functionality of individual gluten fractions along with vari-
2
Department of Food Science &
Technology, College of Agriculture
etal variation in dough and instant noodles quality characteristics, this project was
(Nagaur), Agriculture University, Jodhpur, designed. Glutenins addition enhanced rheological properties such as dough develop-
Rajasthan, India
ment time (DDT), dough stability (DS), and energy at peak (EP) more significantly in
Correspondence DBW88 as compared to HI1500. Gliadins addition amplified dough softening (DSO)
B. S. Khatkar, Department of Food
Technology, Guru Jambheshwar
more prominently in HI1500 than DBW88. Instant noodles cooking and textural qual-
University of Science and Technology, ity improved with glutenins addition and deteriorated with gliadins incorporation.
Hisar, India.
Email: [email protected]
Dough microstructure revealed denser and compact network with extended strands
on glutenins addition, whereas gliadins supplementation produced shorter sheeted
network. Surface microstructure of developed instant noodles displayed smooth con-
tinuous sheet on glutenins incorporation which served as barrier for oil absorption; on
the contrary, gliadins addition created porous and rough surface that ensued higher
oil intake. Thus, strengthening and weakening effect of gluten fractions was eluci-
dated in generating better quality instant noodles.
Practical applications
Selection of wheat variety with appropriate protein content or incorporation of
strength providing gluten fraction in weak variety for improving instant noodles cook-
ing and textural quality with reduced oil uptake.

1 | I NTRO D U C TI O N flours with gliadins and glutenins and alliance of secondary structure
of storage proteins with noodle's textural attributes. Yao et al. (2020)
Instant noodles serve as the solution owing to its palatability, ease studied the varying influence of gluten content on the textural and
of cooking, economic price, shelf stability, and sensory appeal. In inner structure, which revealed that the hardness of the noodle was
terms of global demand for instant noodles, India stands in fourth increased till 14% of gluten content, while nonsignificant difference
place after China/Hong Kong, Indonesia, Vietnam as per statistics existed between the texture of noodles and gluten content when
of World Instant Noodle Association (WINA) (2021). The major and gluten ranged from 8.0% to 11.8%. Therefore, the further influence
prime ingredient is wheat flour combined with minor ingredients of gluten composition for evaluating noodles' texture, cooking qual-
like salt, water, and gums for manufacturing of instant noodles. The ity, and inner structure to establish a relationship with gluten protein
functional components of flour like starch and protein fractions in- fractions is requisite.
fluence dough rheology, thus playing a critical role in the textural and Exploration of different microscopic techniques like confocal
cooking attributes of instant noodles. Therefore, a wise selection of laser scanning microscopy (CLSM), scanning electron microscopy
wheat varieties to manufacture the product with desired quality is (SEM), etc., in depicting the microstructure has been exploited. In
a vital condition (Gulia et al., 2014). Kaur et al. (2015) demonstrated this regard, the influence and interaction of various ingredients
the strong relationships of physicochemical properties of durum and formulations as well as processing techniques on instant

J Food Process Preserv. 2022;00:e17100. wileyonlinelibrary.com/journal/jfpp © 2022 Wiley Periodicals LLC. | 1 of 12

https://doi.org/10.1111/jfpp.17100
2 of 12 | DEWAN et al.

noodles' texture have also been investigated. Using CLSM and gliadins (R 2 = −0.567). The researcher also revealed the relevance
SEM, Qi et al. (2019) analyzed surface properties and oil distribu- of HMW-­GS composition where varieties with 5+10 subunit are
tion in instant noodles and demonstrated the subsequent effects likely to produce the best noodle with firm and nonsticky texture,
of four different frying oils and three emulsifiers on its oil ab- while the varieties with HMW-­GS composition of Null, 2+12, and
sorption. Rise in quinoa proportion along with 40% feed moisture 20 alleles at Glu-­A1, B1, and D1 produced the poor quality of instant
adjustment revealed more smooth surfaces as depicted through noodles with more breakage and rough surface.
microstructure assessment (SEM) of extruded instant noodles Influence of gluten protein fractions on noodle quality in terms
(Tiga et al., 2021). In vitro digestion for fried and nonfried instant of rheology and microstructure has not been explored by prepar-
noodles showed varied honeycomb and smooth microstructural ing instant noodles with isolated gluten fractions. Although forti-
patterns, respectively, on its evaluation by way of SEM (Wang fication of isolated gliadin and glutenin fractions in the dough has
et al., 2022). Thus, recognition to consider microstructural as- been studied (Barak et al., 2014b), but the influence of individual
sessment has been taken care of in this research to have in-­d epth gluten fractions was missing regarding instant noodle quality. In lieu
knowledge about instant noodles. to understand the functionality of individual gluten fractions along
Micro-­doughlab has been a boon, being efficient, and reproduc- with varietal variation in dough and instant noodles quality charac-
ible, while requires a small amount of sample to determine rheological teristics, this project was designed. Therefore, in the present study,
properties, when the base flour has been fortified with gluten and its the effects of isolated gluten fractions on dough rheology and its
subfractions (glutenin, LMW-­GS [low molecular weight glutenin sub- microstructure were assessed. Also, the influence of isolated gliadin
units], gliadin, and its subfractions) (Chaudhary et al., 2022; Dangi and glutenin fractions on instant noodle quality parameters and its
et al., 2019; Khatkar et al., 2013). Researchers have made significant microstructure was studied.
discoveries about the functionality of individual wheat protein frac-
tions and successfully established the relations with the end prod-
uct quality (Barak et al., 2014a; Chaudhary et al., 2016b; Chaudhary 2 | M ATE R I A L S A N D M E TH O DS
et al., 2022). The relationship of gliadin and glutenin fractions was
established in significant correlation with dough rheological param- 2.1 | Flour preparation
eters like dough development time and stability, where a positive
association was found with glutenins, While negative relationship Two different wheat varieties DBW88 and HI1500 were selected
with gliadins (Barak et al., 2013). The role and interaction among the for this research based on its diverse physicochemical characteris-
individual gluten fractions (gliadins, high molecular weight glutenin tics and Glu-­1 score as mentioned in Table 1. Cleaned wheat grains
subunits (HMW-­GS), and low molecular weight glutenin subunits of both varieties were milled into straight grade flour in chopin
(LMW-­GS)) were revealed through the rheological and microstruc- laboratory mill (Model CD1). Flour of both varieties was utilized to
tural analysis (Bonilla et al., 2020). Fractionation and reconstitution extract gliadins and glutenins and as the respective base flours for
of wheat flour gluten protein and its subunit have made a significant fortification.
effect on the functionality of gluten and its fraction in baking per-
formance (Graßberger et al., 2003). Successful relationship of wheat TA B L E 1 Flour quality parameters of strong (DBW88) and weak
fractions such as starch, gluten, and lipids on chapatti quality has (HI1500) wheat varieties
been established through fractionation and reconstitution (Arya
Quality parameters DBW88 HI1500
et al., 2016).
Grain hardness (N) 88.70 90.76
Instant noodle features (oil uptake [OU], color, and textural
Falling number (s) 618 605
properties), as well as noodle dough properties (water absorption
SDS sedimentation volume (ml) 66 47
and color), are influenced by the amount and quality of proteins
in wheat flours. Instant noodles made from wheat flour with high Protein content (%) 12.98 10.50

protein content, high SDS sedimentation volume, and low alcohol Resistance to extension ratio (R/E) 3.34 0.83
and salt-­soluble protein content have a bright color, low-­fat absorp- Dry gluten (%) 11.50 9.50
tion during the frying process, and a firm and elastic cooked noodle Gluten index 97.33 58.66
texture (Park & Baik, 2004). Gluten proteins composed of gliadins Gliadin (g/100 g) 4.6 5.4
and glutenins contribute to the elasticity and viscosity of noodles, Glutenin (g/100 g) 6.9 4.1
respectively. The quantity and quality of gluten protein greatly Gliadin/glutenin ratio 0.66 1.39
influence the dough and noodle quality (Gulia et al., 2014). Park
HMW-­GS composition
et al. (2003) established significant correlations of textural proper-
Glu-­A1 2* Null
ties of cooked noodles with gluten quantity and quality including
Glu-­B1 17+18 20
dough rheological properties, SDS sedimentation volume, and score
Glu-­D1 5+10 2+12
of HMW-­GS. Gulia and Khatkar (2015) highlighted a significant cor-
2 Glu-­1 score 10 4
relation of instant noodle texture with glutenins (R = 0.619) and
DEWAN et al. | 3 of 12

2.2 | Grain and flour quality analysis development time (min), dough stability (min), dough softening
(FU), and peak energy (Wh/kg) were estimated.
Single Kernel Characterization System was used to examine the
hardness of the kernel (SKCS, Model 4100, Perten Instruments). The
protein content and falling number of the wheat flour were evalu- 2.5 | Noodle making
ated using the (AOAC, 2000) internationally recognized method.
The volume of sodium dodecyl sulfate (SDS) sedimentation volume Instant noodles were developed utilizing optimized processing
was determined using the Axford et al. (1979) technique. The resist- conditions and formula by Gulia and Khatkar (2013). The dough
ance to extension ratio was calculated using Kieffer dough and the was prepared by mixing flour with water containing dissolved
TA-­X T plus Texture Analyzer's gluten extensibility rig (Ktenioudaki salts (Kansui and sodium chloride) and guar gum for 4 min in the
et al., 2010). mixer. The prepared dough was sheeted to 3.2 mm thickness and
then rested for 10 min in a zip lock pouch to prevent moisture
loss. Finally, the thickness of dough sheet was made to 1.2 mm
2.3 | Gluten fractionation and rested for 30 min in a zip lock pouch. The sheets were cut
into strands and placed on a steamer tray for 6.4 min at 100°C.
Gluten proteins were separated into gliadins and glutenins by fol- The steamed noodles were deep fried in soybean oil at 142°C for
lowing the modified Osborne (1907) technique. Gluten isolation 2 min and followed by cooling for 15 min. Prepared instant noo-
was accomplished on defatted flour using the glutomatic device dles' samples were wiped to remove excess oil and stored in zip
and the AOAC (2000) technique. Isolated gluten was lyophilized lock pouches for further analysis.
by using a Lark lyophilizer. Fifty grams of powdered gluten was
extracted in 1 L of 70% (v/v) ethanol and mixed for 3 h at room
temperature (22°C) on a magnetic stirrer. The mixture was then 2.6 | Quality evaluation of instant noodles
centrifuged at 1000 g for 30 min at 4°C in a cooling centrifuge.
The extraction was carried out three times. The alcohol insolubles 2.6.1 | Oil uptake (OU)
were collected as glutenins, and the alcohol-­s oluble gliadins were
recovered from the supernatant by evaporating the ethanol with The Soxhlet method was used to determine oil uptake (AOAC, 2000).
a rotary evaporator at 30°C. In a pestle mortar, the freeze-­dried Preweighed thimble with 3 g of ground sample was introduced in a
gliadin and glutenin fractions were pulverized. Gluten content (%), soxhlet assembly for 6 h of petroleum ether extraction (60–­8 0°C).
gliadin (%), glutenin (%), and gliadin/glutenin ratio were calculated Defatted samples were dried and weighed. Oil uptake on dry weight
on dry matter basis after freeze-­drying. basis was determined as follows.

(Initial weight of sample − Final weight of sample)

Oil uptake ( % ) = × 100
2.4 | Dough rheology analysis of dough with added Initial weight of sample

gliadin and glutenin fractions

2.6.2 | Cooking quality (CQ)
A 4-­g Micro-­d oughlab (Perten) was used to assess the effects of
gliadin and glutenin fractions on dough rheological parameters. CQ like cooking time (CT), cooking loss (CL), and cooked weight (CW)
Strong (DBW88) and weak (HI1500) variety flours were added was determined as per Gulia and Khatkar (2013). In a 500-­ml beaker,
with gliadin and glutenin fractions at levels of 3% and 5%, respec- fried noodle stands (10 g) were added to 400 ml boiling water and
tively. 3% and 5% levels were selected on the basis of maximum cook until the disappearance of the inner white core. Drained and air-­
protein percentage existing in the wheat varieties. The isolated dried noodle strands were weighed for calculation of cooked weight
gliadin and glutenin fractions were incorporated into their respec- with mentioned formula indicating the weight gain during cooking. To
tive base flour to evaluate their effects on dough rheology and quantify cooking loss, a portion of 50 ml of the water that remained
end-­p roduct quality. Addition of 3% and 5% gluten proteins was after cooking, along with rinsings, was collected and dried in an oven
added based on preliminary trials. The resulted flour was placed at 105°C for 4 h. The results were expressed as a percentage weight
in the micro-­d oughlab mixing bowl and given dry mix for 1 min decrease during the cooking process.
before adding water to attain the 500 FU force line on the chart.
All of the runs took 15 min, employing the default micro-­d oughlab Cooking loss ( % ) =
Dried residues left after cooking
× 100
Weight of sample taken for cooking
profile. However, because the dough was excessively firm and
the departure time could not be estimated within 15-­min cycle,
the mixing time for the glutenin fortified flours was manually ex- Weight of cooked sample
Cooking weight ( % ) = × 100
tended to 30 min. Rheological properties of dough, such as dough Weight of raw sample taken for cooking
4 of 12 | DEWAN et al.

TABLE 2 Influence of gliadins and glutenins on rheological parameters, oil uptake, cooking quality, and textural properties of instant noodles

Sample DDT (min)* DSO (FU)* DS (min)* PE (Wh/kg)* Oil uptake (%)* Cooking time (min)*

Control (DBW 88) 5.33f ± 0.11 65.00 d ± 0.18 7.53g ± 0.35 10.60 f ± 0.40 17.77d ± 0.16 2.31b ± 0.01
+3% Gliadins 4.10 e ± 0.17 85.00 e ± 0.00 4.27e ± 0.46 6.87e ± 0.11 21.42f ± 0.15 2.44d ± 0.01
c e c cd h
+5% Gliadins 3.50 ± 0.01 90.00 ± 0.00 3.27 ± 0.11 6.23 ± 0.05 25.73 ± 0.41 2.51e ± 0.01
+3% Glutenins 8.33g ± 0.23 −6.67b ± 0.88 11.93h ± 0.28 16.37g ± 0.57 15.33b ± 0.35 4.05i ± 0.01
h a i h a
+5% Glutenins 10.00 ± 0.01 −28.33 ± 0.88 13.03 ± 0.51 17.63 ± 0.45 13.61 ± 0.13 4.16j ± 0.01
Control (HI 1500) 2.73b ± 0.05 56.67d ± 0.88 4.20 d ± 0.17 4.50 b ± 0.00 19.85e ± 0.11 2.05a ± 0.03
a e b a g
+3% Gliadins 2.30 ± 0.01 93.33 ± 0.11 2.63 ± 0.05 3.57 ± 0.20 23.36 ± 0.15 2.41c ± 0.01
+5% Gliadins 2.13a ± 0.05 115.00 f ± 0.00 2.17a ± 0.05 3.53a ± 0.20 29.08j ± 0.15 3.01f ± 0.01
d c e c d
+3% Glutenins 3.73 ± 0.05 35.00 ± 0.00 5.43 ± 0.05 6.13 ± 0.23 17.52 ± 0.56 3.41g ± 0.01
+5% Glutenins 4.13e ± 0.05 25.00 c ± 0.00 7.03f ± 0.35 6.70 de ± 0.00 15.97c ± 0.18 4.01h ± 0.01

Abbreviations: DDT, dough development time (min); DSO, dough softening (FU); DS, dough stability (min); PE, energy at peak (Wh/kg).
*Mean values followed by different letters within a same column differ significantly (p > 0.05) and expressed as mean ± standard deviation.

2.6.3 | Instrumental texture profile analysis sulfate (SDS) sedimentation volume was higher (66 ml) for flour of
DBW88 indicating as strong variety, while variety HI1500 resulted
The textural profile of instant noodles in terms of hardness, springi- in 47 ml, indicating it as a weak variety. Falling number is a meas-
ness, adhesiveness, cohesion, and chewiness was analyzed employ- ure for the amylase activity, which was 618 s for variety DBW88
ing texture analyzer (Stable micro Systems TA-­X T 2i) as per Gulia and and 605 s for variety HI1500. Grain hardness was more for variety
Khatkar (2013) method. HI1500 than variety DBW88. Gluten index signifies the strength
and quality of gluten protein as the strong and elastic gluten will not
pass through the sieve when subjected to centrifugation, whereas
2.7 | Microstructural analysis weak gluten will easily pass through the sieve. The higher value of
gluten index for variety DBW88 indicated higher gluten strength
For microstructural analysis of freeze-­dried dough and instant noo- with 97.33, whereas variety HI1500 displayed weaker gluten with
dles, fractured samples were fixed on carbon tape and pasted on 58.66 gluten index. This was supported by the gluten composition
the sample stub. The exposed fractured area of sample was coated where the presence of a higher proportion of glutenins (6.9%) and
with gold nanoparticles using sppurtek coater (DIT-­290305 CIR). lower proportion of gliadins (4.6%) in variety DBW88, while vice
The coating was done for 0.5 s with a thickness of 2–­3 nm to make versa for variety HI1500. Similarly, Dangi et al. (2019) also worked
the entire sample conductive. These gold-­coated samples were ana- on two wheat varieties C306 and PBW550, where the former had
lyzed on scanning electron microscope (JSM-­7610F Plus JEOL) with higher gliadin content (5.7%) and lower glutenin content (3.8%),
EDAX-­APEX software. The micrographs of freeze-­dried dough were exhibited the weaker gluten and dough strength, while the latter
viewed at a magnification of 2000× and for instant noodles at 100×. one with a higher proportion of glutenins (5.4%) and lower propor-
tions of gliadins (4.2%) unfolded the stronger gluten and dough
rheology. The HMW-­GS composition of variety DBW88 expressed
2.8 | Statistical analysis subunits 2*, 17+18, and 5+10 with Glu-­1 score 10, while variety
HI1500 expressed subunits Null, 20, and 2+12 with Glu-­1 score 4.
The data were subjected to anova using Duncan's multiple range test Zhao et al. (2020) have proven that the genotypes with 5+10 subu-
for statistical significance, and correlation employing SPSS software nit have shown to exhibit strong dough rheology and good baking
version 16.0 (SPSS Inc.). performance.

3 | R E S U LT S A N D D I S CU S S I O N 3.2 | Dough rheological properties

3.1 | Grain and flour analysis The effect of gliadins and glutenins addition in the base flour at
3% and 5% levels on dough rheological properties is presented
The grain and flour analysis of varieties DBW88 and HI1500 is in Table 2. An increase of gliadin content reduced the DDT from
summed up in Table 1. Varieties DBW88 and HI1500 had protein 5.33 to 3.50 min for variety DBW88, while for variety HI1500,
contents of 12.98% and 10.50%, respectively. The sodium dodecyl DDT was reduced from 2.73 to 2.13 min. However, a significant
DEWAN et al. | 5 of 12

Cooking weight Adhesiveness

(g/100 g)* Cooking loss (%)* Hardness (N)* Springiness* Cohesiveness* Chewiness (N)* (N/s)*

235.60 f ± 2.02 4.52d ± 0.07 39.35f ± 0.92 0.97c ± 0.04 0.86b ± 0.03 32.82c ± 0.21 −0.13f ± 0.02
246.67g ± 0.18 4.99e ± 0.01 36.94e ± 0.88 0.94c ± 0.03 0.82b ± 0.04 28.47c ± 0.12 −0.77d ± 0.10
h f c b b b
250.44 ± 0.02 5.20 ± 0.01 32.10 ± 0.96 0.81 ± 0.03 0.80 ± 0.05 20.80 ± 0.59 −1.45b ± 0.01
198.90 b ± 0.30 3.63b ± 0.01 40.90 g ± 0.91 1.08e ± 0.07 1.16d ± 0.02 51.24f ± 0.07 −0.02g ± 0.01
a a h f d g
187.56 ± 0.33 3.18 ± 0.05 44.84 ± 0.64 1.20 ± 0.06 1.20 ± 0.04 64.57 ± 1.15 −0.01g ± 0.01
222.23e ± 1.24 6.16g ± 0.08 32.92c ± 0.80 1.06de ± 0.01 0.99c ± 0.01 34.55c ± 1.02 −0.21e ± 0.00
gh h b a a a
248.12 ± 1.99 6.54 ± 0.09 30.34 ± 1.02 0.71 ± 0.05 0.70 ± 0.05 15.07 ± 0.59 −1.18c ± 0.01
259.73i ± 1.08 6.92i ± 0.11 27.85a ± 0.58 0.71a ± 0.05 0.69a ± 0.01 13.64a ± 0.82 −1.63a ± 0.05
d c d cd d d
213.29 ± 1.62 3.98 ± 0.01 34.93 ± 0.43 1.01 ± 0.04 1.19 ± 0.02 41.98 ± 0.19 −0.08fg ± 0.01
207.82c ± 0.46 3.24a ± 0.05 38.23ef ± 0.57 1.02cde ± 0.01 1.19d ± 0.04 46.25e ± 1.75 −0.03g ± 0.01

increment was revealed with a rising percentage of glutenins from of HI1500+5% glutenins) in comparison with control and dough
5.33 to 10.00 min in variety DBW88 and from 2.73 to 4.13 min supplemented with gliadins. The reason lies in the protein's na-
in variety HI1500. The time difference between the arrival time ture, that is, gliadins are monomeric viscous proteins while glu-
and departure time of the curve in the mixograph of flour is de- tenins are polymeric elastic proteins forming the gluten network
fined as dough stability (DS). Decline in DS was observed from in the dough. Gliadins fortification resulted in developing soft
7.53 min in control to 3.27 min in the dough with 5% gliadins for and weak dough may be due to the absence of cysteine residues
DBW88 variety, while in the case of variety HI1500, pronounced as exhibited by Chaudhary et al. (2022) revealed the decline in
decline was noticed from 4.20 min in control to 2.17 min in the DS and DDT with higher values for DSO through the rheological
dough with 5% gliadins. This was clearly observed that increasing analysis of isolated pure gliadins and its subfractions. Strong and
gliadin proportion made the dough less stable, by virtue of its vis- dense gluten association was created in dough supplemented with
cous and fluidic nature. Whereas DS was increased with increased glutenins due to its extensive intermolecular disulfide linkages
concentration of glutenins, due to the development of more intra-­ (Chaudhary et al., 2016a). The inclusion of gliadins and glutenins
and inter-­disulfide bonds. This was supported by the researchers in variety DBW88 showed pronounced effects as compared to va-
(Oszvald et al. (2011), Dangi et al. (2019)) stating large amount of riety HI1500.
mixing energy requirement and dough stability was observed with
the supplementation of polymeric wheat proteins (i.e., glutenins)
in the dough. DS surged significantly from 11.93 to 13.03 min with 3.3 | Microstructure of dough
the rising percentage of glutenins in the flour of DBW88 from
3% to 5%, respectively. While in variety HI1500 at 3% and 5% The effect of gliadins and glutenins supplementation on the dough
levels of glutenins addition, DS was raised to 5.43 and 7.03 min, microstructure was studied using SEM micrograph as shown in
respectively. This may be due to the difference in its high mo- Figure 1. On 3% gliadin fortification in base flour of variety DBW88,
lecular weight glutenin composition and their number of cysteine an open gluten network with short strands and broken small sheets
residue, considering variety DBW88 comprised of subunit 5+10 holding starch granules were observed. Furthermore, increasing
and variety HI1500 with 2+12 as subunit. The presence of extra gliadins addition to 5% displayed the appearance of shorter sheets
cysteine residue in the 5+10 subunit (Wieser, 2007) is account- enfolding starch molecules with a more increased number of cavi-
able for the dough strength, therefore requiring more time to de- ties. As a result, the appearance of hollow gaps due to the fluidic and
velop dough by virtue of creating more disulfide bonds resulting viscous nature of globular gliadins was noted, leading to the shorten-
in greater dough stability. Similar results have been observed by ing of the gluten network. The strong dough characteristics of va-
Barak et al. (2013) and Dangi et al. (2019) where wheat varieties riety DBW88 were further enhanced with glutenin fortification at
PWB550 and PBW590 with 5+10 subunit exhibited strong dough the levels of 3% and 5%, where in the former case, thin long strands
rheology. The quantity of energy required by dough to reach its with continuous sheets of gluten network were clearly visible, while
maximum height is denoted by energy at peak (EP). Strong dough in the latter case, increased larger and uninterrupted gluten network
requires a large amount of energy than weak dough. The dough sheets were visible entrapping the starch molecules. The possible
supplemented with glutenins showed the highest EP (17.63 Wh/ reason for dough strength is glutenin's nature and structure, which is
kg for flour of DBW88+5% glutenins and 6.70 Wh/kg for flour a polymeric protein comprising cysteine residues capable of forming
6 of 12 | DEWAN et al.

DBW88 control DBW88 + 3% gliadins DBW88 + 5% gliadins DBW88 + 3% glutenins DBW88 + 5% glutenins

HI1500 control HI1500 + 3% gliadins HI1500 + 5% gliadins HI1500 + 3% glutenins HI1500 + 5% glutenins

F I G U R E 1 Micrographs of control flour dough of DBW88 and HI1500 wheat varieties along with 3% and 5% of gliadin and glutenin
fractions.

inter-­and intra-­disulfide bond leading to the formation of a well-­ with good and poor bread-­making quality, Dhaka and Khatkar (2016)
structured dense gluten network, thus serving as the backbone of revealed the structure of gluten and its fraction (gliadins and glute-
the gluten (Khatkar et al., 2013). nins) with micrographs and they produced a more ordered network
By 3% fortification of gliadins in the base flour of HI1500, which of glutenins and gluten of strong variety HI977. Furthermore, the
is natively composed of higher gliadin content with low Glu-­1 score, author's results were coinciding with gliadins behavior in C306 (poor
resistance extension ratio, and SDS sedimentation volume devel- bread-­making quality) forming self-­assembled sheet like network
oped short sheets of the gluten network. An increase in the quan- and acting as plasticizer in gluten association as it was observed in
tity of shorter film-­like structures covering the starch granules was micrographs of HI1500 dough. The enhanced concentration of glu-
discovered when the percentage of gliadins surged from 3% to 5%. tenins created a more uniform, dense, and closely intact dough net-
HI1500 base flour fortified with 3% and 5% of glutenins content work. Inclusion of purified gliadin fractions was shown to produce
showed a visible larger sheet structure with the continuous network shorter strands, thus resulting in the open gluten structure due to its
with short strands but in comparison with strong variety DBW88, monomeric nature and property of providing fluidity to the devel-
sheets were comparatively smaller in HI1500 dough structure. oped dough (Chaudhary et al., 2022; Dewan et al., 2022).
With the increased proportion of gliadins in the strong variety Starch association with gliadins and glutenins in dough occurs
DBW88, the presence of excelling monomeric molecules created through chemical interaction (ionic bonds, hydrogen bonds, disul-
more hollow spaces in the network was remarked, while in weak fide bonds as well as hydrophobic interactions). Hydrophobic in-
flour of HI1500, continuous sheeted matrix was revealed. The teractions come into force due to the presence of nonpolar amino
strong variety was predicted to entrap the gliadins monomeric pro- acids of gluten molecules. When gluten is hydrated, the formation
tein within its intense gluten matrix (due to more glutenins content), of unstable hydrogen bond forces takes place between hydro-
whereas variety HI1500 with additional gliadin proteins resulted in philic amino acids present on the surface of gliadins or glutenins,
a more sheeted network due to increase in the viscosity. As a result, or hydroxyl groups free from starch granules (Li et al., 2021). On
the native dough created a more open and weak gluten network, increasing the proportion of gliadins to glutenins, hydrophobic
suggestively due to its viscous nature and globular morphology as interaction decreases. This is because glutenin being fibrous in
conferred by Khatkar et al. (2013). The inclusion of glutenins in both structure has many repetitive hydrophobic regions existing on
the varieties developed a more continuous, strong, compact, and the surface of the molecule, and gliadin is spherical in morphol-
more well-­organized dough matrix conferring the dough strength- ogy with many hydrophobic amino acid side chains exposed on the
ening. Similar results were observed by Dewan et al. (2022) and surface of the molecule (Markgren et al., 2020). Glutenins form
Dangi et al. (2019) where flour was fortified with glutenins depicting an organized fibrous macromolecular polymer through disulfide
a strong and compact gluten matrix. The results were strengthened bonding, where starch molecules are embedded through nonco-
by the microdough lab values, where glutenins supplementation to valent bonding in the ordered glutenin network. While gliadins are
both the varieties depicted greater dough stability and strength. monomeric proteins that interact through intramolecular disulfide
More continuous and dense structure was observed in variety bonding and a large number of hydrophobic amino acid residues
DBW88, while in variety HI1500, few cavities and hollow gaps were located on the spherical interior. Gliadins and starch granules get
observed, which highlighted the difference of glutenin subunit com- embedded in the fibrous network of glutenin polymer leading to
position of both the varieties. To differentiate between the varieties the formation of a loop-­like structure through intra-­chain covalent
DEWAN et al. | 7 of 12

and noncovalent bonds. With an increasing proportion of gliadin a less permeable outer layer, which could act as an efficient barrier
content, gliadins compete with starch to absorb water, forming against oil absorption.
phase separation or adsorption of gliadin on the surface of starch CT was observed to be maximized with the increase in the
(Li et al., 2021). protein content, although the result revealed a significant in-
crease from 2.31 to 4.16 min and 2.05 to 4.01 min in both variet-
ies DBW88 and HI1500, respectively. Significant increment was
3.4 | Oil uptake (OU) and cooking quality (CQ) of observed in glutenins addition as compared to gliadins due to the
instant noodles development of tough and strong gluten network by polymeric
wheat protein fraction requiring more time for cooking. Increment
Instant noodles that were prepared from varieties DBW88 and in CW was observed from 235.60% (control) to 250.44% (5 g of
HI1500 along with 3% and 5% fortification of gliadins and gluten- gliadins/100 g flour) in variety DBW88 and from 222.23% (con-
ins in their respective flour are shown in Figure 2. These prepared trol) to 259.73% (5 g of gliadins/100 g flour) in variety HI1500 with
instant noodles were subjected to OU and CQ evaluation summa- increasing gliadin content. CW declined with increasing glutenin
rized in Table 2. The instant noodles prepared from control dough % from 198.90% (3 g of glutenin/100 g flour) to 187.56% (5 g of
of variety DBW88 were found to have 17.77% OU, which escalated glutenin/100 g flour) for variety DBW88, while 213.29% (3 g of
from 21.42% with 3% gliadins to 25.73% with 5% gliadins addition. glutenin/100 g flour) to 207.82% (5 g of glutenin/100 g flour) for
OU in instant noodles prepared from control of variety HI1500 was variety HI1500. Inclusion of glutenin proteins creates a confined
19.85%, which elevated from 23.36% to 29.08% with gliadin con- and rigid network that is responsible for resisting the water up-
tent varying from 3% to 5%, respectively. Oil absorption raised with take of instant noodles during cooking, while with increased gli-
increasing proportion of gliadin content but on comparing both adin content, an open porous structure resulted in more water
the varieties, HI1500 natively comprising more gliadin content ex- absorption. This might be the possible reason for higher CW with
hibited higher OU than DBW88 comprising more glutenin content increased gliadin proportion, while vice versa for glutenins in-
natively. Gliadins created extensible dough with a more open and clusion. For variety DBW88, CL increased from 4.99% to 5.20%
porous structure resulting in a rough surface, which causes higher with a rise in gliadins from 3% to 5% in the native flour, whereas
oil absorption. Inclusion of glutenin fraction in the dough at 3% and declined from 3.63% to 3.18% with surging glutenins from 3% to
5% decreased the oil absorption from 15.33% to 13.61% in DBW88, 5% on compared with 4.52% (control). While a similar pattern was
while the same pattern of decline in OU from 17.52% to 15.97% with observed in HI1500, but the values were significantly higher due
3% and 5% of glutenins inclusion was revealed in HI1500. Reduction to the varietal and basic compositional difference in gluten com-
in oil absorption with surging glutenin percentage in both varieties position. Chaudhary et al. (2016b) findings imply that polymeric
was apparently due to the enhanced proportion of polymeric glu- proteins network with starch polymer slows the hydration of noo-
tenin fractions responsible for the dough strength and elasticity, dles by reducing the amount of water available during cooking and
thus, creating a smooth outer surface serving as a barrier for oil ab- lengthening the cooking time. Furthermore, the water diffusion
sorption (Gulia & Khatkar, 2015; Moreno et al., 2010). According to ability during the rehydration and cooking process is influenced
Kaur et al. (2015), a stronger gluten network is responsible to create by noodle porosity offering the framework for water to infiltrate

DBW 88 (control) DBW 88 + 3% gliadins DBW88 + 5% gliadins DBW88 + 3% glutenins DBW88 +5% glutenins

HI 1500 (control) HI1500 + 3% gliadins HI1500 + 5% gliadins HI1500 + 3% glutenins HI1500 +5% glutenins

F I G U R E 2 Instant noodles prepared from control flour of DBW88 and HI1500 along with 3% and 5% gliadin and glutenin fractions.
8 of 12 | DEWAN et al.

into the middle of the noodles (Jang, Kim, & Lee, 2016). In gen- and chewiness (15.07–­13.64 N, respectively), while no difference
eral, soluble constituents of wheat flour like starch, pentosans, was revealed with springiness values (0.71). The varietal differ-
and water-­s oluble protein fractions like albumin, and globulins are ences in the textural values were due to the native composition of
lost during the cooking of instant noodles. In the results, cooking the varieties as HI1500 has an initial higher proportion of gliadins
loss was increased with increasing gliadin content. This is because as compared to the glutenins exhibiting a weakening effect on the
gliadin incorporation leads to the loosening and opening of the textural attributes of instant noodles, whereas DBW88 exhibited
gluten network (Khatkar et al., 2013). Additionally, gliadin incorpo- strong instant noodles textural properties due to the higher per-
ration tends to alter glutenin–­gliadin ratio which in turn adhered to centage of glutenin proteins present in the native flour. Gliadins
the gluten network causing its weakening and it is easy for water viscous and fluid kind nature leads to the development of weak
molecules to invade and dissolve the soluble constituents (Cao soft dough and pliable noodles, while glutenin fortified dough and
et al., 2021). Therefore, noodle porosity is dependent on the com- noodles were strong and firm. The result was supported by the
position of the flour and the network created by the protein and study of Gulia and Khatkar (2015) where firm and elastic instant
starch interaction deciding the oil absorption and cooking quality noodles were prepared by wheat varieties with a higher propor-
of the developed noodles. It can be concluded that glutenins are tion of glutenin content, while low yield with poor textural quality
responsible for creating a strong gluten network, therefore, en- of instant noodles was produced by wheat varieties with a greater
hancing the cooking quality of instant noodles with less oil absorp- content of gliadin proteins.
tion, while gliadins incorporation results in weak and open gluten Adhesiveness is defined by the amount of force required to sep-
structure responsible for the deterioration of its cooking quality arate the noodles sticking in the mouth after chewing which is not
along with more OU. desirable as nonsticky noodles are preferred. Gliadins incorporation
increased the adhesiveness from −0.77 N/s (3% gliadins in DBW88)
to −1.45 N/s (3% gliadins in DBW88) and −1.18 N/s (3% gliadins in
3.5 | Textural characteristics of noodles HI1500) to −1.63 N/s (5% gliadins in HI1500), while decline from
−0.02 N/s (3% glutenins in DBW88) to −0.01 N/s (5% glutenins in
The textural parameters of cooked instant noodles are compiled DBW88) and −0.08 N/s (3% glutenins in HI1500) to −0.03 N/s (5%
in Table 2. The hardness of the cooked instant noodles prepared glutenins in HI1500) with increasing glutenin content. Varieties with
from DBW88 variety surged with glutenins incorporation from strong dough rheology and high glutenin content give nonsticky
40.90 N at 3% to 44.84 N at 5% level, while declined with gliadins noodles, while the varieties with high gliadin content give stickier
fortification from 36.94 N at 3% to 32.10 N at 5% level on compari- noodles. Noodles with a firm and elastic structure were produced
son with control 39.35 N. However, in variety HI1500, hardness employing strong wheat varieties like DBW16 and HI977 with higher
hiked from 34.93 N with 3% glutenins to 38.23 N with 5% gluten- glutenin and lower gliadin content, while varieties like HW2004 and
ins, but dropped down on gliadins addition from 30.34 N at 3% to C306 with weak dough rheology generated inferior quality noodles
27.85 N at 5% level on comparing with 32.92 N (control). Increase which were stickier and breaking due to a weak network (Chaudhary
in the hardness was observed in both the varieties with the in- et al., 2016b).
corporation of glutenin fraction. Noodles are produced by sheet-
ing, which is regulated by the adequate proportion of gliadins and
glutenins in the flour. The presence of higher proportion of glu- 3.6 | Microstructure of instant noodles
tenins responsible for elasticity will result in recoiling of noodle
sheets, while higher gliadins creating greater extensibility will lead Researchers have explored the microstructure of dough and noo-
to tearing or rough surface development (Gulia & Khatkar, 2015). dles to understand the interaction of ingredients and its composi-
Plasticizing effect of gliadins produced soft noodles, which was tion along with the effect of processing conditions on the noodle
also marked by Barak et al. (2014a); Gulia and Khatkar (2015). quality like OU, cooking, and textural quality using SEM (Gulia
Researchers observed that the wheat varieties with a higher pro- et al., 2014). The micrographs of instant noodles (Figure 3) are de-
portion of gliadins produced fragile noodles, while with greater picting the functional role of gliadins and glutenins in the starch
glutenins, harder noodles were produced. gluten network, which thereby are responsible for the oil intake
The same pattern was observed in other textural attributes mechanism, textural, and cooking quality. Micrographs of instant
like springiness, cohesiveness, and chewiness accounted by the noodles (gliadins were incorporated at the level of 3% and 5%)
instant noodles network created by specific protein owing to made from a variety DBW88 revealed the cavity-­like structure in-
its nature. With the enhancement in gliadins addition from 3% creased and more hollow spaces were observed. Whereas on the
to 5% in instant noodles prepared using DBW88, the decline in addition of increasing glutenins from 3% to 5%, denseness in the
springiness (0.94–­0 .81, respectively), cohesiveness (0.82–­0 .80, network with an enhanced number of strands with the increase in
respectively), and chewiness (28.47–­20.80 N, respectively) was the thickness was visible indicating a more bonded and compact
observed. Similarly, gliadins fortification from 3% to 5% in HI1500 structure. Nie et al. (2019) revealed the continuity and integrity of
resulted in decrement in cohesiveness (0.70–­0 .69, respectively) the gluten network comprising of gluten strands and gluten film
DEWAN et al. | 9 of 12

'%:FRQWURO '%:JOLDGLQV '%:JOLDGLQV '%:JOXWHQLQV '%:JOXWHQLQV

+,FRQWURO +,JOLDGLQV +,JOLDGLQV +,JOXWHQLQV +,JOXWHQLQV

F I G U R E 3 Micrographs of instant noodles prepared from control flour of DBW88 and HI1500 along with 3% and 5% fortification of
gliadin and glutenin fractions.

binding the starch granules. Glutenin fortified samples were ob- 3.7 | Correlation among rheological characteristics,
served with the continuous dense network of polymeric proteins oil uptake, cooking, and textural attributes of
with starch resulting in hard, firm, springy, and cohesive noodles instant noodles prepared with gliadin and glutenin
depicted by textural parameters in Table 2. HMW-­G S are proven fortification
to form the backbone of gluten network, which are present in
wheat glutenins responsible for strength of the dough and thereby Correlation among rheological and instant noodle quality char-
strong dense network results in a decrease in OU and CL as pre- acteristics like oil uptake (OU), cooking time (OU), cooking loss
sented in Table 2. Pongpichaiudom and Songsermpong (2018) (CL), cooked weight (CW), and textural attributes is summed up in
revealed the matrix of instant dried noodles where large starch Table 3. Strong significance was revealed among the dough rheol-
granules were entrapped by protein strands forming a network by ogy and instant noodle quality parameters. DDT produced highly
disulfide, hydrogen, and hydrophobic bonds, which diluted on with significant positive correlation with DS (R 2 = 0.97), EP (R 2 = 0.99),
the fortification of meat proteins leading to a more porous struc- hardness (R 2 = 0.91), and chewiness (R 2 = 0.85) at 99% significance
ture. However, Nawaz et al. (2019) elucidated that the starch–­ level. DS also exhibited a well-­b uilt correlation with EP (R 2 = 0.97),
protein matrix with compact, dense, and rigid microstructure with hardness (R 2 = 0.93), and chewiness (R 2 = 0.92) at the 99% sig-
fewer voids leads to less oil uptake in comparison with a structure nificance level. This indicated that the dough rheological proper-
comprising more intracellular spaces between the starch and pro- ties symbolizing the dough strength is in great coherence with
tein network responsible for higher oil absorption during frying. the textural properties of instant noodles. Polymeric wheat pro-
Micrographs of instant noodles prepared from HI1500 man- teins, which are responsible for dough strengthening, lead to the
ifested that there was a decline in the continuity of the network, development of strong, firm, cohesive, and springy instant noo-
with gliadins fortification. A few shorter strands with more open dles. Similar results were revealed by Yang et al. (2020) through
cavities were observed in the micrograph of the HI1500 variety with establishing a relationship among dough rheological parameters
5% gliadins when compared with the HI1500 control. Continuity in and noodles' quality. Tanδ which is the indicator of the balance
sheeted structure with fewer cavities and the hollow gap was ob- between viscous and elastic nature of dough was found in great
served in the micrographs of the dough of variety HI1500 with 3% coherence with the hardness of the noodles with R 2 = 0.786 at
and 5% glutenins. Gliadins supplemented samples showed more cav- 95% significance level. Other rheological features like R% (defor-
ity and open-­sheeted network in instant noodles resulting in more mation degree), η × 105 Pa.s (coefficient of viscosity), σmax × 10 4 Pa
OU and CL. Furthermore, textural properties of gliadins fortified (maximum stress), ε (flow behavior index), and KPa.s (consistency
noodles revealed soft and adhesive noodles due to the viscous and coefficient) also exhibited a strong positive correlation with the
monomeric characteristics of gliadin proteins. The effect of gliadins noodles textural properties like springiness, cohesion, and chewi-
addition on dough microstructure has been clarified by Chaudhary ness. The author also displayed that the noodles quality param-
et al. (2022) where the open network with short strands resulted eters such as optimal cooking time, water absorption, CL, and
from gliadins addition in the dough resulting in a soft and extensible breaking length were significantly in the positive influence of
gluten network. G′Pa (storage modulus) and G″Pa (loss modulus), while negatively
 10 of 12
|

TA B L E 3 Correlation among dough rheological, cooking quality, and textural parameters of instant noodles

Hardness Chewiness Adhesiveness

Correlation DDT DSO DS PE OU CT CW CL (N) Springiness Cohesiveness (N) (N/s)

DDT 1.00
DSO −0.86** 1.00
DS 0.97** −0.93** 1.00
** **
PE 0.99 −0.83 0.97** 1.00
OU −0.68* 0.87** −0.81** −0.66* 1.00
CT NS −0.54 NS NS NS 1.00
CW −0.78** 0.99** −0.87** −0.74* 0.89** −0.56 1.00
* ** ** * *
CL −0.75 0.85 −0.81 −0.72 0.72 −0.60 0.82** 1.00
Hardness 0.91** −0.86** 0.93** 0.90** −0.78** NS −0.81** −0.88** 1.00
* ** * * ** ** **
Springiness 0.76 −0.90 0.83 0.73 −0.79 NS −0.91 −0.78 0.86** 1.00
** * ** * ** *
Cohesiveness 0.53 −0.86 0.69 0.50 −0.87 0.69 −0.91 −0.70 0.57 0.73* 1.00
** ** ** ** ** ** ** **
Chewiness 0.85 −0.98 0.92 0.81 −0.86 0.55 −0.98 −0.88 0.80 0.94** 0.84** 1.00
** * ** ** ** ** ** **
Adhesiveness 0.58 −0.82 0.72 0.56 −0.90 NS −0.86 −0.77 0.78 0.89 0.81 0.86** 1.00

Abbreviations: DDT, dough development time (min); DSO, dough softening (FU); DS, dough stability (min); PE, energy at peak (Wh/kg); OU, oil uptake (%); CT, cooking time (min); CW, cooking weight (%); CL,
cooking loss (%); NS, nonsignificant.
**Correlation is significant at the 0.01 level (two-­t ailed).; *Correlation is significant at the 0.05 level (two-­t ailed).
DEWAN et al.
DEWAN et al. | 11 of 12

correlated with R% and tanδ. Thus, it was discovered that higher prominent in HI1500 than DBW88 due to the higher proportion of
values of G′, G″ with the lower value of tanδ displayed strong gliadins present in the former one. Instant noodles CQ enhanced
elastic behavior due to the presence of glutenin macropolymer re- with least CW, CL, and oil absorption with the supplementation of
sulting in high-­quality noodle texture as well as cooking behavior. glutenins, while vice versa was observed with gliadins supplementa-
Similar results were obtained in this research, where instant noo- tion. The supporting evidences were derived from SEM results of
dles' cooking parameters such as CW and CL were found in strong dough and instant noodles. Thus, fractionation and fortification of
negative correlation with hardness (R 2 = −0.81 and R 2 = −0.88, re- gliadins and glutenins at 3% and 5% in the native dough assisted in
spectively), springiness (R 2 = −0.91 and R 2 = −0.78, respectively), the dough rheology and instant noodles quality in terms of its oil
cohesiveness (R 2 = −0.91 and R 2 = −0.70, respectively), chewiness absorption, cooking, textural quality, and its microstructure.
2 2 2
(R = 0.98 and R = −0.88, respectively), DDT (R = −0.78 and
R 2 = −0.75, respectively), DS (R 2 = −0.87 and R 2 = −0.81, respec- C O N FL I C T O F I N T E R E S T
tively), and EP (R 2 = −0.74 and R 2 = −0.72, respectively). These The authors have declared no conflicts of interest for this article.
correlations signified the inverse relation of solid loss and weight
gain of cooked instant noodles with strength denoting textural DATA AVA I L A B I L I T Y S TAT E M E N T
and dough characteristics, which are directly proportional to the All the data used in the manuscript are available in the figures and
strength providing gluten fractions. It might be due to the strong tables
compact dense structure built by glutenins holding starch mol-
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