Effects of water-unextractable arabinoxylan from wheat processing wastewater on the quality characteristics of multigrain bread

Prévia do material em texto

Effects of water-unextractable arabinoxylan from wheat processing 
wastewater on the quality characteristics of multigrain bread
Sihai Han a,b, Yanbin Wang a, Yunhui Zhang a, Honglin Lan a, Xingguo Li a, Jiahao Wan a, 
Chonghui Yue a, Junwei Feng c, Denglin Luo a,b, Zhouya Bai a,b,*
a College of Food and Bioengineering, Henan University of Science and Technology, 471023, Luoyang, Henan, China
b Henan Food Raw Material Engineering Technology Research Center (Henan University of Science and Technology), Luoyang, 471023, China
c Henan Feitian Biotechnology Co., Ltd., Hebi, 456750, China
A R T I C L E I N F O
Keywords:
Arabinoxylan
Wheat processing wastewater
Water-unextractable arabinoxylan
Substitution
Functional bread
A B S T R A C T
The water-unextractable arabinoxylan (WUAX) was extracted from the wheat processing wastewater to replace 
the flour in the bread formula and improve the quality characteristics of the multigrain bread. We investigated 
the effects of WUAX substitution concentration (0, 2.5, 5, 7.5 and 10 g/100 g high gluten flour weight) on 
multigrain bread texture, microstructure, color difference variation, relative crystallinity, moisture distribution, 
and sensory evaluation. Within the specified range, the bread hardness increased by 4.0 N, a negative effect on 
the bound water in bread and significant changes in the color difference were observed. The optimal substitution 
level was 7.5%, increasing the crude fiber content by 33% (dry weight ratio) and reducing the starch content by 
10%, maintaining the specific volume and hardness similar to the control group. The scanning electron micro-
scopy X-ray diffraction (XRD) measurements showed that WUAX affected the aggregation of protein and starch, 
and reduced the relative crystallinity of bread, retarding bread staling. The WUAX replacing flour (7.5% 
replacement level) also provided good sensory evaluation. Overall, WUAX boosts bread’s dietary fiber content 
while lowering its starch proportion. Although WUAX had negatively impact on bread texture, a 7.5% substi-
tution level could minimize these effects. This study offered insights for the dietary applications of WUAX from 
wheat processing wastewater.
1. Introduction
The history of wheat cultivation in China can be traced back to the 
Neolithic period. With the continuous improvement of varieties and the 
promotion of ancient rulers, wheat has had a profound influence on 
China’s agricultural industrial structure, people’s diet structure and 
culture. In the process of wheat processing, a large amount of high- 
concentration organic wastewater is produced from the washing, pres-
sure filtration, concentration and other process steps. The wastewater 
contains a lot of soluble organic matter, such as starch, sugar, fat, amino 
acids, inorganic substances, etc (Bai & Lan, 2024; Geng and Li et al., 
2024). Among them, arabinoxylan (AX) is a highly researched 
non-starch polysaccharide, which has been widely used as nutritional 
supplement to regulate the quality of flour products (Li and Li et al., 
2023).
Arabinoxylan mainly consists of a 1,4-linked pyran xylose residue 
backbone with random O-2, O-3, or O-2 and O-3 arabinose substitutions, 
and can be classified as water-extractable arabinoxylan (WEAX) and 
water-unextractable arabinoxylan (WUAX) (Pandeirada and Merkx 
et al., 2021). The content of WUAX in wheat processing wastewater is 
much greater than that of WEAX, and has the possibility of extraction in 
large quantities. The extraction of WUAX from wheat processing 
wastewater offers a unique opportunity for both environmental reme-
diation and economic benefits (Li, Li & et al, 2023), and could develop a 
source of valuable dietary fiber for various functional food applications 
(Asif & Khan, 2014).
Bread is a staple food around the world, and has become a symbol of 
cultural diversity and culinary craftsmanship. Each country and region 
boast its own unique bread variety, reflecting local climate, agricultural 
conditions, and historical-cultural influences that have shaped the 
people’s taste preferences (Gao & Zhou, 2021). In recent years, the 
functionally fortified breads have received increased interests. There 
have been many novel functional breads, such as moringa seeds fortified 
bread (Bolarinwa and Aruna et al., 2019), bee pollen bread (Conte and 
* Corresponding author. College of Food and Bioengineering, Henan University of Science and Technology, 471023, Luoyang, Henan, China.
E-mail address: spbaizhouya@163.com (Z. Bai). 
Contents lists available at ScienceDirect
LWT
journal homepage: www.elsevier.com/locate/lwt
https://doi.org/10.1016/j.lwt.2024.116867
Received 31 July 2024; Received in revised form 24 September 2024; Accepted 5 October 2024 
LWT - Food Science and Technology 210 (2024) 116867 
Available online 9 October 2024 
0023-6438/© 2024 The Authors. Published by Elsevier Ltd. This is an open access article under the CC BY-NC-ND license ( http://creativecommons.org/licenses/by- 
nc-nd/4.0/ ). 
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Del et al., 2018), olive pulp bread (AlJuhaimi and Ahmed et al., 2024), 
and bread with sheep whey powder (Secchi and Fadda et al., 2018). 
With the rise of health consciousness, consumers are paying more 
attention to the nutritional value of bread. Whole-grain bread, enriched 
with fiber and essential nutrients, has become a popular choice (Mir and 
Farooq et al., 2022). The baking process has also undergone innovations 
to reduce the energy content while maintaining the flavor and texture 
(Kikuchi and Nozaki et al., 2018). Furthermore, the combination of 
traditional baking methods with modern technology has given birth to a 
range of innovative bread products. From gluten-free options for those 
with dietary restrictions to low-sugar and high-fiber varieties for 
health-conscious consumers, the possibilities are endless.
AX is a natural dietary fiber with many bioactivities such as regu-
lating intestinal microbiota (Paesani et al., 2020), reducing risk of type 2 
diabetes (Pereira et al., 2021) and enhancing immunomodulation 
(Zhang, Yang, Zhou, Shen, & Hu, 2021). In recent years, AXs have been 
applied to improve the quality of flour baking products (Bieniek & 
Buksa, 2024). It was reported that WEAX could improve the viscoelas-
ticity of gluten in dough while WUAX could induce fragile discontinuous 
structure of gluten (Li & Li et al., 2023). It was also reported that both 
WEAX and WUAX could impart positive influence on dough properties 
by increasing the water absorption capacities of flours and enhancing 
the stability of dough, and that WEAX could improve physical and 
sensory attributes of bread while WUAX exhibited negative effects (Arif, 
Ahmed, Chaudhry, & Hasnain, 2018). However, WUAX was a valuable 
dietary fiber with high content and availability, and could provide many 
health benefits such as regulating short-chain fatty acid in the colon, 
enhancing antioxidant capacity, and reducing blood glucose response 
(Huang & Bai et al., 2024).
Most studies about effects of WUAX on bread properties were per-
formed using the WUAX extracted from cereals, and few of them 
employed the WUAX from wheat processing wastewater. Therefore, the 
objective of this work is to evaluate the feasibility of fortifying a func-
tional bread with WUAX isolated from wheat processing wastewater. 
The effect of WUAX on multigrain bread texture, microstructure, color 
difference variation, moisture distribution, staling properties, as well as 
sensory evaluation, was investigated, which provided a scientific sup-
port for the application of WUAX in baking products.
2. Materialsand methods
2.1. Preparation of WUAX
The WUAX with particle size less than 100 mesh was extracted from 
wheat processing wastewater (produced by Henan Feitian Agriculture 
Development Co., Ltd., Hebi City, China). The wheat processing 
wastewater was centrifuged at 4000 rpm for 15 min, and the sediment 
was obtained. The WUAX was extracted from the sediment with an 
alkaline solution (pH 12) at 65 ◦C for 3 h. The extraction solution was 
adjusted to pH 7, standing for 2 h, then was centrifuged to obtain the 
supernatant. Subsequently, the WUAX was purified from the superna-
tant by condensing (to 1/20 of original volume) with rotary evapora-
tion, dialyzing at 4 ◦C for 24 h, and vacuum freeze-drying (− 20 ◦C for 24 
h) in sequence.
2.2. Bread making process
The high gluten flour and red kidney bean powder were purchased 
from Yihai Wheat Co., Ltd. (Henan, China). The butter was purchased 
from Jinshan Branch of Inner Mongolia Yili Industrial Group Co., Ltd 
(Inner Mongolia, China). The sugar (purity 99.5%) was purchased from 
Rizhao Lingyunhai Sugar Co., Ltd (Nanjing, China). The active dry yeast 
was purchased from Anqi yeast Co., Ltd (Hubei, China).
The WUAX was used instead of 2.5%, 5%, 7.5% and 10% of wheat 
flour (dry basis, w/w) for bread making, respectively. The blank control 
group was made using the same bread formula without the WUAX 
substitution. The base formulation of bread included 100g of high gluten 
flour, 20 g of egg mixture, 10 g of red kidney bean powder, 10 g of broad 
bean flour, 10 g of oat flour, 10 g of buckwheat flour, 10 g of gluten 
flour, 8 g of butter, 10 g of white sugar, 1.8 g of dry yeast and 60 g of 
water.
The formulation ingredients were mix thoroughly and kneaded for 
20 min with a multifunctional chef machine (PE4680, Guangdong Liran 
Electric Appliance Industry Co., Ltd., Guangdong, China). The dough 
transferred into molds for a brief relaxation period, following a 
fermentation at 37 ◦C and 80% relative humidity for 90 min in a 
fermentation chamber (FJX-16, Guangdong Demas Intelligent Kitchen 
Equipment Co., Ltd. Guangdong, China). After fermentation, the molds 
with dough were baked in a preheated (160 ◦C) oven (SM 522, Xinmai 
Machinery Co., Ltd.) and baked with temperature 150 ◦C (top and bot-
tom) for 10 min.
2.3. Determination of specific volume and porosity of bread
The specific volume of bread samples (cm3/g) was determined using 
the reference method (Wang and Chen et al., 2017). The bread core part 
with uneven pores in the bread was not used as the determination 
sample. The Image J software was used for the bread picture processing 
and the data calculation (Han and Liu et al., 2024). The results were 
presented as images and porosity values.
2.4. Color analysis
The color difference measurement referred to literature 
(Iglesias-Puig & Haros, 2013) with minor modifications. A square bread 
sample (20✕20✕20 mm3) was taken from the center of the bread core. 
The color values were measured using a colorimeter (X-rite color i5, 
USA) with a pulse xenon lamp illuminant and di:8◦ measurement ge-
ometry. The color values determined included L*, a* and b* 
(García-Hernández and Roldan-Cruz et al., 2023), which represent 
brightness (0 for black, 100 for white), red-green degree (-a for green, 
+a for red), and yellow-blue degree (-b for blue, +b for yellow), 
respectively. The total color change (ΔE) was calculated using formula 
(1). 
ΔE=
̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅
(ΔL*)2
+ (Δa*)2
+ (Δb*)2
√
(1) 
2.5. Texture profile analysis (TPA)
The texture characteristics of the bread were determined by the 
professional texture analyzer (TA.XT. Express, Stable Micro Systems, 
UK) equipped with a P/75 probe. to ensure the stable and reliable re-
sults. The bread sample was cut into a square (20✕20 ✕20 mm), and 
determined with trigger force of 5 g, compression ratio 50%, early 
extrusion rate of 12 mm/s and 2 mm/s in middle-late stages (Li & 
Gidley, 2022; Liu, Liu, Huang, & Zhang, 2021). The interval of extrusion 
was 1 s. The results were expressed in terms of hardness, springiness and 
other data.
2.6. X-ray diffraction (XRD) analysis
Grilled freeze-dried bread samples were analyzed using an X-ray 
diffractometer. The sample powder was tested at 40 mA, 40 kV, and 
room temperature. Diffraction angle ranged from 5◦ to 40◦, scanned at 
5◦/minute. The crystal properties of the peak and amorphous regions 
were analyzed using the Origin software, and the relative crystallinity 
was calculated using the crystal formula TC = lc/(lc + la). The results 
were presented as crystallinity data.
S. Han et al. LWT 210 (2024) 116867 
2 
2.7. Moisture mobility and distribution determination (LF-NMR)
The water distribution of the bread was determined using a low-field 
nuclear magnetic resonance instrument (LF-NMR, NMI 20-015 V-I, 
Shanghai Newmarket Electronics Technology Co., Ltd., China). The in-
strument mainly determines the moisture content of three different 
binding states by measuring the lateral relaxation time of the bread. The 
proportion of bound water, weakly bound water, and free water was 
determined. Experiments were performed at 32 ◦C, with each sample 
weight set to 1.5 g, and placed evenly in a glass bottle with a sampling 
frequency of 200 kHz and a sampling interval of 3500 ms. The results 
were expressed in terms of relaxation time and inverse peak area of NMR 
images.
2.8. Scanning electron microscopy (SEM) analysis
The protein and starch microstructure of bread were studied by 
electron scanning microscopy (TM3030Plus desktop scanning electron 
microscope: Hitachi, Japan). The bread was cut into large pieces and 
then sprayed with gold. Images were taken at a magnification of 400 and 
1000. The results were shown as SEM images.
2.9. Analysis of physicochemical properties of bread
The protein, fat, crude fiber, starch content and water absorption 
rate of bread were analyzed. Protein content was determined by Kjeldahl 
method (Qi and Li et al., 2013). Chloroform methanol measurement was 
used to measure fat (Elliott & Elliott, 2016). The determination of crude 
fiber was carried out using an acid-based solution method, taking 
advantage of the insolubility of cellulose in weak acids and bases (Taus 
and Tahuk et al., 2022). The starch content was measured with anthrone 
colorimetry kit (Nanjing Jiancheng Bioengineering Institute). Water 
absorption was analyzed as described in literature (Liu, Liu, & Huang 
et al., 2021). The results were presented in bar charts.
2.10. Sensory evaluation of bread
The sensory evaluation of bread samples was performed by a panel of 
10 trained panelists (5 males and 5 females, mean ages: 20–55 years) 
complying with National Bread Sensory Evaluation Standard GB/T 
20981-2021 of China (Han and Liu et al., 2024). All the panelists were 
from the College of Food and Bioengineering, Henan University of Sci-
ence and Technology. The informed consent from the participants was 
obtained. The sensory evaluation was carried out in a spacious and clean 
room with 360 Lx brightness, relative humidity 75%, and temperature 
25 ◦C. The bread was cut into slice (1 × 1 × 2 cm) and placed in a white 
porcelain plate randomly encoded. The order of tasting was random. 
Mouth rinsing was performed after each test and at 5-min intervals. The 
panelists were asked to rate and give score for different sensory pa-
rameters including shape (15 points), crust color (15 points), crumb 
color (15 points), texture (20 points), flavor/taste (20 points), impurity 
(15 points). The results were presented with the sum of the sensory 
evaluation score.
2.11. Statistical analysis
All experiments were conducted in triplicate, and the results were 
reported as mean ± standarddeviation (SD). Correlation and signifi-
cance analyses of the indicators were performed using SPSS software. 
The significance of the difference between control and treatment sam-
ples was assessed using the least significant difference (LSD) at pof crude WUAX substitution on the structural characteristics of bread.
Hardness 
(N)
Springiness Gumminess Chewiness Resilience
Control 8.3394 ±
0.21c
0.92 ±
0.09a
416.99 ±
27.87d
395.99 ±
9.78c
0.22 ±
0.01b
WUAX 
2.5%
11.4179 
± 0.30ab
0.88 ±
0.07ab
675.08 ±
16.25c
590.54 ±
29.89 b
0.22 ±
0.00b
WUAX 
5%
10.5035 
± 0.94b
0.82 ±
0.00ab
823.15 ±
32.09a
708.14 ±
49.27a
0.20 ±
0.00c
WUAX 
7.5%
10.7555 
± 0.87b
0.86 ±
0.00ab
632.05 ±
17.30c
544.94 ±
17.25b
0.24 ±
0.00a
WUAX 
10%
12.3233 
± 0.36a
0.80 ±
0.01b
750.40 ±
39.20b
591.68 ±
42.18b
0.22 ±
0.01bc
The data were expressed mean ± standard deviation (SD) (n = 3). Means within 
a row with different superscript letters are significantly different (pwas high. Both T22 and T21 achieved a minimum at a substitution value 
of 7.5%. Compared with the control group, all the A21 (indicating the 
content of bound water) of breads with WUAX addition showed a 
downward trend, indicating that WUAX had a negative effect on the 
binding water in bread. The A22 (indicating the weakly bound water 
content) values had no significant difference between the WUAX sub-
stitution group and the control. With the WUAX addition increased, the 
A23 (indicating the content free water) showed a growth trend, and had a 
minimum growth at 7.5% substitution level. Compared with the other 
substitution proportions, the 7.5% WUAX substitution level had the least 
negative effect on the moisture distribution in bread.
3.5. Effect of WUAX addition on bread microstructure
The microstructure of the bread samples with WUAX and without 
WUAX was analyzed by scanning electron microscopy (SEM) compari-
son. SEM micrographs of bread cores of compound bread supplemented 
with different proportions of WUAX are shown (Fig. 4). Compared with 
the control, the breads with WUAX addition had rough edges, local 
structure destruction and debris. This was due to the entanglement of 
gluten protein molecules, which hinders the formation of gluten 
network. WUAX has long molecular chains and many branches that may 
wrap around the gluten structure and organize it to form larger gluten 
polymers. As the substitution amount of WUAX increases, the ability of 
starch to bind between proteins decreases, because the high molecular 
weight of WUAX leads to a large steric hindrance, forming a physical 
barrier to the starch exposed outside of the gluten structure (Ling, Wang 
et al., 2017). In order to more clearly observe the differences between 
bread structures, SEM micrographs of the same dough magnified by 
1000× were shown. It could be seen that there were more cracks in the 
bread after the increase of substitution, and the surface became rougher 
and irregular, and some filamentous structures and discontinuous gluten 
networks appeared. This indicated that the substitution of WUAX 
destroyed the gluten network structure and made the starch exposed, 
which leaded to the poor quality of the bread.
3.6. Analysis of physicochemical properties of bread
Starch content is an important indicator of the energy source of food. 
Food with higher starch content will generally have a higher glycemic 
index, which is unfavorable for those who need to control blood sugar. 
In bread with WUAX substitution (2.5%, 5%, 7.5%, 10% substitution for 
flour), the starch content is much less than that of high-gluten flour 
bread. When the WUAX substitution level was 10%, the starch content 
reached the lowest value, which was about 12% lower than that of the 
control group (Fig. 5). The amount of dietary fiber in bread increased 
with WUAX substitution and reaching a minimum at 10%, about 35% 
higher than that in the blank control group. The protein content also 
decreased, achieving the lowest value at 10%, which was about 3% 
lower compared to the blank control group. There were no significant 
changes in fat content or water absorption. Water absorption could 
reflect the quality characteristics of bread from the side. The addition of 
AX increased the water absorption of bread (Koegelenberg & Chim-
phango, 2017). De Bondt and Hermans et al. studied the addition of 
modified bran to bread and found that bran addition caused a greater 
effect than water absorption (De Bondt and Hermans et al., 2021). The 
effect of WUAX addition was greater than the effect of water absorption 
on bread quality. Natal and de Souza Dantas et al. investigated the effect 
of adding whole soybean flour to potato bread on its physicochemical 
properties and found that the content of protein, dietary fiber, and the 
minerals calcium, zinc, magnesium, copper, and phosphorus increased 
as the concentration of soybean flour increased (Natal and de Souza 
Dantas et al., 2013). The physicochemical data of the bread changed 
mainly because of the different levels of nutrients contained in the added 
functional ingredients. The water absorption rate of WUAX substituted 
breads did not change significantly compared to the control group. 
Generally speaking, the stronger water absorption, the stronger the 
ability of bread to maintain its own combined water, the better the 
quality of bread (Schopf & Scherf, 2021).
The starch in bread is mainly derived from the high gluten flour used 
in the formula, and the starch content can be effectively reduced by 
replacing the high gluten flour with WUAX in bread. The main purpose 
is to reduce the fast digestion of starch in the bread. Fast-digestible 
Table 3 
Effect of crude WUAX on the lateral relaxation time (T2) and peak area (A2).
Group T21 T22 T23 A21 A22 A23
Control 0.21 ±
0.02ab
4.04 ±
0.00a
75.65 ±
0.00a
16.76 ±
0.36a
72.52 
± 0.35a
10.45 ±
0.32c
WUAX 
2.5%
0.20 ±
0.02b
3.86 ±
0.30ab
75.65 ±
5.69ab
16.16 ±
0.69a
71.97 
± 0.58a
11.32 ±
0.35b
WUAX 
5%
0.21 ±
0.02ab
4.24 ±
0.35a
75.65 ±
0.00a
15.76 ±
0.71b
72.04 
± 1.07a
11.85 ±
0.22a
WUAX 
7.5%
0.19 ±
0.00b
3.51 ±
0.00b
65.79 ±
0.00b
14.56 ±
0.63ab
73.15 
± 0.65a
11.53 ±
0.06ab
WUAX 
10%
0.23 ±
0.02a
4.04 ±
0.00a
75.65 ±
5.69ab
15.59 ±
0.79ab
72.10 
± 0.28a
11.77 ±
0.16ab
The data were expressed mean ± standard deviation (SD) (n = 3). Means within 
a row with different superscript letters are significantly different (pcertain health 
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	Effects of water-unextractable arabinoxylan from wheat processing wastewater on the quality characteristics of multigrain bread
	1 Introduction
	2 Materials and methods
	2.1 Preparation of WUAX
	2.2 Bread making process
	2.3 Determination of specific volume and porosity of bread
	2.4 Color analysis
	2.5 Texture profile analysis (TPA)
	2.6 X-ray diffraction (XRD) analysis
	2.7 Moisture mobility and distribution determination (LF-NMR)
	2.8 Scanning electron microscopy (SEM) analysis
	2.9 Analysis of physicochemical properties of bread
	2.10 Sensory evaluation of bread
	2.11 Statistical analysis
	3 Results and discussion
	3.1 Effect of adding WUAX on bread specific volume, color and texture
	3.1.1 Bread specific volume and porosity
	3.1.2 Color attributes
	3.2 Texture profile analysis (TPA)
	3.3 Crystal structure and relative crystallinity
	3.4 Moisture mobility and moisture distribution in bread
	3.5 Effect of WUAX addition on bread microstructure
	3.6 Analysis of physicochemical properties of bread
	3.7 Sensory evaluation
	4 Conclusion
	CRediT authorship contribution statement
	Declaration of competing interest
	Acknowledgments
	datalink4
	References