foods

Article
Influence of Adding Chinese Yam (Dioscorea opposita
Thunb.) Flour on Dough Rheology, Gluten Structure,
Baking Performance, and Antioxidant Properties
of Bread
Qing-Ming Li 1,2 , Yan Li 1 , Jin-Hao Zou 1 , Shi-Yin Guo 1 , Feng Wang 1,2 , Peng Yu 3 and
Xiao-Jun Su 1,2, *
1 College of Food Science and Technology, Hunan Agricultural University, Changsha 410128, China;
[email protected] (Q.-M.L.); [email protected] (Y.L.); [email protected] (J.-H.Z.);
[email protected] (S.-Y.G.); [email protected] (F.W.)
2 Hunan Provincial Research Center of Engineering and Technology for Fermented Food,
Changsha 410128, China
3 College of Science, Hunan Agricultural University, Changsha 410128, China; [email protected]
* Correspondence: [email protected]; Tel.: +86-731-8467-3522




Received: 5 February 2020; Accepted: 24 February 2020; Published: 28 February 2020


Abstract: Impacts of wheat flour substituted with various levels of Chinese yam (Dioscorea opposita
Thunb.) flour (from 0% to 25%) on the dough rheological characteristics, gluten structure, baking
performance, and antioxidant properties of bread were investigated. The water absorption increased
significantly (p < 0.05), while development time and stability decreased remarkably (p < 0.05) as the
proportion of yam flour increased. SEM results indicated that the addition of yam flour destroyed
the gluten network structure in the dough. Fourier Transform Infrared Spectroscopy (FTIR) spectra
showed that addition of yam flour decreased the content of α-helix and β-sheet in gluten. With
the increase in the proportion of yam flour, the specific volume and overall acceptability decreased
(p < 0.05) whereas the total phenolics content (TPC), polysaccharides content, total flavonoids content
(TFC), allantoin content, The 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging capability,
fractal dimension, and hardness increased (p < 0.05). Overall, breads made of wheat flour replacement
with no more than 15% Guihuai number 2 yam flour were of a high quality and had more antioxidant
properties. These showed that Guihuai number 2 had broad application prospects in baked products.

Keywords: yam; bread; gluten structure; antioxidant property; rheology

1. Introduction
Chinese yam (Dioscorea opposita Thunb.) has long been considered a food that improves health.
In China, it is a traditional Chinese herbal medicine. Due to its high protein and starch content,
yam is an important staple food in many subtropical and tropical regions and is the main source of
carbohydrates and proteins [1]. Fresh yams are perishable and not easy to store. Processing of yam
tubers into flour is the main mean to increase its economic value. Yam flour can be used as a food
ingredient, which will broaden the utilization of yam in food processing. Yam flour has been used to
prepare many traditional foods, such as amara, noodles, pasta, and baby food [2–4]. Yam flour has
outstanding potential for making bread as a result of its high nutritional value and good processing
properties [5–8]. Compared with wheat flour, yam flour is abundant in polysaccharide, phenolic,
flavonoid, and allantoin compounds, and has high antioxidant ability [9,10]. Previous studies have
reported that yam and wheat notably affect the functional properties and quality of bread varied with

Foods 2020, 9, 256; doi:10.3390/foods9030256 www.mdpi.com/journal/foods

Foods 2020, 9, 256 2 of 15

the level of substitution. Hsu et al. [6] and Liu et al. [11] reported that wheat substitution with yam
flour resulted in bread with high antioxidant ability and low digestibility. Hsu et al. [6] concluded
that breads made with no more than 20% yam flour did not have significant negative effects on the
bread quality. Gluten makes an important impact on bread making, but yam flour lacks gluten [12].
Understanding the influence of yam flour on the structure of gluten is necessary to optimize the recipe
and process of yam bread. However, there is no report on how the addition of yam flour affects the
changes in gluten structure.
Guihuai number 2 is one of the yam cultivars selected by Cash Crops Research Institute, Guangxi
Academy of Agricultural Sciences (Nanning, China) which has been widely planted for food use in
south China. Compared with other yam cultivars, Guihuai number 2 has the characteristics of high
yield, small starch molecular weight, high amylose content, and low pasting temperature. Guihuai
number 2 has not been fully utilized and well-studied compared to other yam cultivars. Evaluation
of Guihuai number 2 for bread making is beneficial to broaden applications and thus promote the
cultivation of yam. Therefore, it is necessary to evaluate the potential applicability of Guihuai number
2 as a partial substitution for wheat flour in bread making.
Thus, the objectives of this study were to evaluate the effect of bread making substituted with
Guihuai number 2 yam flour. The dough rheological properties and gluten protein structure were
studied using farinograph, extensograph, rheometer, rapid visco analyzer (RVA), texture analyzer,
scanning electron microscopy (SEM) and fourier transform infrared spectroscopy (FTIR). In addition,
the bioactive components and antioxidant properties of bread were also investigated.

2. Materials and Methods

2.1. Materials
Wheat flour was produced by Guangzhou Grain Enterprise Group (Guangzhou, China).
The composition analysis results as following (dry basis): 13.34% ± 0.04% moisture, 14.20% ± 0.10%
protein, 69.87% ± 0.43% starch, 1.70% ± 0.05% fat, and 0.73% ± 0.02% ash content. Fresh yams (Guihuai
number 2) were harvested from Baise city, Guangxi province. The yam tubers were washed, peeled,
and sliced to 3mm thickness pieces. Yam slices were dried in a 101-2AB hot air dryer oven (Tianjin
Taisite Instrument Co., Ltd, Tianjin, China) at 60 ◦ C [13]. After drying, the slices were pulverized using
a hammer (SF-130, Zhongnan Pharmaceutical Machinery Factory, Changsha, China) and sieved via a
mesh of 250 µm. The proximate composition of yam flour was as following (dry basis): 6.77% ± 0.06%
moisture, 12.76% ± 0.13% protein, 62.55% ± 0.33% starch, 0.56% ± 0.03% fat, and 3.95% ± 0.07% ash
content. Salt, sugar, instant dry yeast (Angel Yeast Co. Ltd, Chengdu, China), and shortening (Anchor,
Auckland, New Zealand) were purchased from METRO supermarket in Changsha, China. All other
chemicals and reagents were of analytical grade unless otherwise stated.

2.2. Dough Preparation and Laboratory Bread Making Method

The method described by Nindjin, Amani, and Sindic [7] was used in bread making. The bread
recipe was described as follows: 100 g of wheat flour, 0.7 g of instant dry yeast, 8 g of shortening, 18 g
of sugar, 1 g of salt, and 55 g of water. Wheat flour was substituted with 0% (W100Y0), 5% (W95Y5),
10% (W90Y10), 15% (W85Y15), 20% (W80Y20), or 25% (W75Y25) yam flour weight.

2.3. Dough Properties

2.3.1. Farinograph and Extensibility

Farinograph and extensibility were performed on farinograph and extensograph (Brabender E,
Duisburg, Germany) respectively, with the method described by Li [3] and Nindjin [7].
Foods 2020, 9, 256 3 of 15

2.3.2. Texture Properties

The dough was cut into 50 × 40 × 15 mm cubes. Texture properties of dough were measured using
texture profile analysis (TPA) on a TA.XT plus texture analyzer (Stable Micro Systems, Surrey, UK) with
a P100 probe. The detailed condition was described in a previous study with slight modification [14].
Measurement conditions: the pre-test speed (2.0 mm/s), test and post-test speed (1.0 mm/s), and
strain (65%).

2.3.3. Rheological Analysis

Dough rheological analysis was performed on a Kinexus Pro+ rheometer (Malvern Instruments,
Worcestershire, UK). Measurement conditions: parallel-plate geometry (20 mm diameter) with 1 mm
gap, frequency range of 0.1–20 Hz and a temperature of 25 °C. Furthermore, elastic modulus (G0 ),
viscous modulus (G”), and loss tangent (tan δ = G”/G0 ) were also determined [14].

2.3.4. Pasting Properties

The pasting analysis was performed on RVA (Model: RVA-S/N2112681, Perten, Stockholm,
Sweden). The method was presented in previous reports [15,16]. Mixed flour samples (3.5 ± 0.01 g)
were transferred into the canister where 25 mL of distilled water was added. The suspension was
heated from 50 to 95 ◦ C at a rate of 12 ◦ C/min, maintained at 95 ◦ C for 2.5 min, followed by cooling to
50 ◦ C at a rate of 12 ◦ C/min with another 2 min holding time.

2.4. Structure Analysis of Dough and Gluten

2.4.1. Drying of Dough and Gluten

Water-washed gluten was prepared by using the procedure of Tuhumury et al. [17]. The wet
gluten and dough were freeze-dried using a freeze-dryer (FD-1-50, Beijing Biocool Experimental
Instrument Co. Ltd, Beijing, China).

2.4.2. Scanning Electron Microscopy (SEM)

Micrographs of dough and gluten were visualized by a JSM-6380LV SEM (JEOL Co., Tokyo, Japan).
The freeze-dried dough and gluten samples were cut into 10 × 10 × 10 mm cubes. The samples were
fixed, coated with sputtered gold and examined under SEM at an energy of 25 kV.

2.4.3. Fourier Transform Infrared Spectroscopy (FTIR)

FTIR measurements of gluten were performed on an IRAffinity-1 spectrometer (Shimadzu, Kyoto,
Japan). The absorbance spectra was recorded between 4000–400 cm−1 , at a resolution of 4 cm−1 , and
32 scans. Baseline correction was performed on the amide I band (1600–1700 cm−1 ) using peakfit
4.12 software (SeaSolve Software Inc., Framingham, MA, USA). The Savitsk–Golay function was
used for smoothing and deconvolution. The relative percentage of each secondary structure was
calculated [18].

2.5. Evaluation of Bread Quality

2.5.1. Bread Image Analysis

Bread was cooled at room temperature for 2 hours and then cut into 12.5 mm thick slices.
The crumb images were taken with a color camera. The crumb characteristics were analyzed by the
method described by Gao et al. [19] and Farrera-Rebollo et al. [20].
Foods 2020, 9, 256 4 of 15

2.5.2. Specific Volume (SV)

Cooled bread was accurately weighed, and volume was determined by the millet replacement
method. The specific volume (mL/g) was calculated from the ratio of volume and mass of bread.

2.5.3. Crumb Texture

Crumb texture was performed on a TA-XT2 texture analyzer (Stable Microsystems, Surrey, UK).
Strain was 70% [21], other conditions were the same as Section 2.3.2.

2.5.4. Antioxidant Property

Moisture content was determined by heating sample at 105 ◦ C for 4 hours in a 101-2AB hot air dryer
oven (Tianjin Taisite Instrument Co. Ltd., Tianjin, China). Fresh bread was used for test and the extract
methods were described by Hsu et al. [1,6]. Total polyphenol content (TPC) and total flavonoid content
(TFC) were analyzed according to the analytical procedure described by Chiu [9] and Chen [22], the
results were expressed as mg gallic acid equivalent (GE)/g and rutin equivalent (RE)/g of bread sample
(dry basis ), respectively. Allantoin was determined on an Agilent 1260 HPLC with a C18 column
(150 mm × 4.6 mm, 5 µm). The conditions were described by Fu, Ferng and Huang [10]. Measurement
of polysaccharides content was conducted on the basis of the method reported by Yang et al. [23].
The results were expressed as mg/g of bread sample (dry basis). The 1,1-diphenyl-2-picrylhydrazyl
(DPPH) scavenging activity of bread was measured using the procedure described by Hsu et al. [6].
The EC50 value represented the effective concentration when 50% of radicals were scavenged.

2.5.5. Sensory Evaluation

The sensory panelists (5 males and 5 females; age 20–40) were recruited based on their previous
experience in the sensory evaluation of baked foods among postgraduate students and teachers of the
Hunan Provincial Research Center of Engineering and Technology for Fermented Food, China. Sensory
evaluation was determined according to a nine point hedonic rating scale method, the analytical
procedure as described by Nindjin [7].

2.6. Statistical Analysis

All trials were carried out in triplicate and the data expressed by average ± standard deviation
(SD) were analyzed by SPSS 22.0 (IBM, Chicago, IL, USA) and statistical significance was determined
at 0.05 levels (p ≤ 0.05).

3. Results and Discussion

3.1. Effect of Yam Flour Substitution on Pasting, Farinograph, Rheological, and Texture Properties of Dough
The dough properties were evaluated using RVA, farinograph, extensograph, and texture analyzer,
and the results were listed in Tables 1 and 2. Rheological properties were closely related to the
baking ability of the dough, which was an important index for evaluating the baking performance.
Substitution of wheat flour with yam flour had remarkable effects on pasting properties (Table 1).
Substitution of yam flour led to a remarkable (p < 0.05) decrease of trough viscosity (from 1540 to 350 cP
(1 cP = 10−3 Pa·s)), final viscosity (from 2737 to 745 cP), peak viscosity (from 2275 to 1285 cP), peak time
(from 6.60 to 5.02 min) and setback (from 1198 to 395 cP), while the overall trend of breakdown showed
remarkable (p < 0.05) increase. The decrease of viscosity values could be related to the composition,
starch structure and mucilage of yam. Similar trends were reported that the viscosity values were
significantly decreased by purple yam flour [3], purple sweet potato flour [24], and soybean flour [25]
addition. An increase in breakdown indicated a decrease in the thermal stability of the mixed flour,
which may be related to the amylose content. Addition of yam flour decreases the setback of the mixed
Foods 2020, 9, 256 5 of 15

flour (p < 0.05). This is due to amylose–amylopectin ratio changes in these mixed flours, which had
impact on the crumb structure and the specific volume of bread [14].

Table 1. Effects of yam flour addition on pasting, farinographic, and extensographic properties of
mixed flour.

Item W100Y0 W95Y5 W90Y10 W85Y15 W80Y20 W75Y25

Pasting temperature (◦ C) 69.10 ±0.05b 70.02 ±0.88ab 69.97 ±0.88ab 69.42 ± 0.51b 70.02 ±0.80ab 71.12 ± 1.0a
Peak time (min) 6.60 ± 0.07a 5.93 ± 0.07b 5.58 ± 0.08c 5.38 ± 0.04d 5.31 ± 0.03d 5.02 ± 0.02e
Peak viscosity (cP) 2275 ± 52.92a 1441 ± 6.66b 1271 ± 18.33c 1285 ± 7.00c 1232 ± 2.31d 1285 ± 9.54c
Trough viscosity (cP) 1540 ± 52.92a 629 ± 9.45b 440 ± 13.53c 391 ± 13.23d 355 ± 1.73d 350 ± 5.51d
Final viscosity (cP) 2737 ± 54.60a 1488 ± 9.02b 1068 ± 12.17c 911 ± 23.12d 791 ± 10.02e 745 ± 25.51e
Setback (cP) 1198 ± 15.56a 859 ± 17.44b 628 ± 5.00c 520 ± 10.69d 436 ± 9.61e 395 ± 20.30f
Breakdown (cP) 736 ± 43.50e 811 ± 7.51d 831 ± 25.51c 894 ± 9.17b 877 ± 0.58b 935 ± 8.08a
Water absorption (%) 63.4 ± 0.00d 63.6 ± 0.06c 64.5 ± 0.10b 65.4 ± 0.06a 65.5 ± 0.10a 65.4 ± 0.10a
Dough development time (min) 2.9 ± 0.06a 2.2 ± 0.06b 1.9 ± 0.10c 1.7 ± 0.00d 1.7 ± 0.06d 1.7 ± 0.06d
Dough stability (min) 18.1 ± 0.20a 15.3 ± 0.35b 15.0 ± 0.20b 1.5 ± 0.06c 1.5 ± 0.10c 1.5 ± 0.06c
Degree of softening (FU) 22 ± 0.58e 35 ± 1.53d 44 ± 2.52c 86 ± 2.65a 76 ± 3.06b 88 ± 2.00a
Area under the curve (energy, cm2 ) 114 ± 4.58a 82 ± 3.06c 93 ± 4.00b 68 ± 1.00d 66 ± 4.16d 58 ± 2.52e
Extensibility (mm) 108 ± 5.03a 89 ± 5.69b 105 ± 6.11a 83 ± 3.78bc 80 ± 4.51bc 76 ± 2.00c
Resistance to extension (EU) 878 ± 20.52a 843 ± 25.36a 831 ± 30.62a 736 ± 31.90b 713 ± 22.05b 634 ± 44.96c
Max-extensible resistance (EU) 944 ± 46.52a 858 ± 28.10b 844 ± 23.90b 737 ± 15.52c 733 ± 14.53c 658 ± 31.97d
Extensible rate 8.16 ± 0.35bc 9.53 ± 0.57a 7.90 ± 0.18c 8.91 ± 0.25ab 8.90 ± 0.64ab 8.35 ± 0.48bc
Max-extensible rate 8.77 ± 0.38ab 9.69 ± 0.30a 8.04 ± 0.68b 8.93 ± 0.58ab 9.15 ± 0.69a 8.67 ± 0.60ab
The values (mean ± standard deviation) followed by the same letter in the same row were not significantly different
(p > 0.05) as determined by Duncan’s multiple range test (n = 3).

Table 2. Effects of yam flour addition on texture properties of mixed flour dough.

Sample Hardness (g) Adhesiveness (g·s) Chewiness (g)

W100Y0 4891.46 ± 282.38c 5129.16 ± 398.46d 3932.19 ± 226.28c
W95Y5 3919.22 ± 157.88d 5328.83 ± 242.85d 3121.39 ± 193.17d
W90Y10 5084.16 ± 121.99c 5016.86 ± 301.85d 4031.96 ± 144.53c
W85Y15 5712.79 ± 475.73b 5709.71 ± 86.72c 4543.46 ± 318.80b
W80Y20 6722.81 ± 334.47a 7440.66 ± 351.81a 5551.42 ± 239.90a
W75Y25 5537.48 ± 178.36b 6356.22 ± 246.05b 4622.98 ± 189.89b
The values (mean ± standard deviation) followed by the same letter in the same column were not significantly
different (p > 0.05) as determined by Duncan’s multiple range test (n = 3).

The farinographic analysis could reflect the rheological properties of the dough during its
formation [26]. After adding yam flour, farinograph properties of dough changed significantly (p < 0.05).
According to Nindjin et al. [7], yam starch caused a decrease in water absorption, development time
and dough stability. On the contrary, this result showed that increasing the addition of yam flour,
the water absorption was increased. Similar findings were observed in purple yam flour [3], potato
flour [27]. The increase in water absorption of the mixed flour may be due to the yam flour containing
a highly absorbent mucilage. Development time of the dough reflects the quantity and quality of
gluten. The higher the quantity and quality of gluten, the longer the development time. Development
time and dough stability of different yam flour proportions were remarkably decreased. The softening
degree of W100Y0 was the lowest, which was increased significantly with yam flour addition. Because
gluten is not present in yam flour, mixed flour dough could not develop a strong gluten network,
which reduced the cohesivity and elasticity of the dough. Liu [27] reported that water absorption and
degree of softening increased with the increasing addition of potato flour in mixed flour (10%, 15%,
20%, 25%, 30%, and 35%).
The effect of yam addition on extensographic properties after 135 min was also outlined in Table 1.
The area under the curve, extensibility, resistance to extension, and max-extensible resistance of the
dough showed a significant downward trend with increasing yam flour proportion from 0% up to
25% (p < 0.05). According to Li et al. [3], wheat flour substituted with 5%–20% purple yam flour had
an increased resistance to extension and decreased extensibility. This may be related to the different
Foods 2020, 9, 256 6 of 15

varieties of yam. Energy area was the total energy demanded when the dough was broken down, and
it was an important parameter that reflects the elasticity of the dough. A higher energy area means
more dough elasticity [28]. The addition of yam flour reduced the gluten content of the yam–wheat
mixed flour, resulting in a decrease in the content of gliadin and glutenin, leading to a reduction
in viscoelasticity.
As showed in Table 2, In case of W95Y5 and W90Y10, the adhesiveness was similar to W100Y0
(p > 0.05). Compared with W100Y0, hardness and chewiness of W95Y5 were largely reduced (p < 0.05).
Hardness, adhesiveness and chewiness of dough showed a significant tendency for increasing first
and then decreasing with the addition of yam flour from 5% to 25% (p < 0.05). Similar trends were
also reported by Belghith Fendri et al. [29]. The higher strength of dough with yam flour addition
demonstrated that the bread obtained presented higher hardness.
The frequency sweep test was conducted to analyze the rheological properties of dough. Figure 1
showed the elastic modulus (G0 ), viscous modulus (G”), and loss tangent (tan δ) of the dough. G0
was larger than G” (tan δ < 1) at frequency range from 0.1 to 20 Hz. These phenomena indicated that
elastic properties were predominated and the dough had solid, elastic-like features. The G0 and G” of
all dough samples increased as frequency increased. Similar trends were reported by Cao [30] and
Foods 2020,
Balestra 9, x FOR PEER REVIEW
[31]. 7 of 15

120 Elastic modulus G′

100

80
G'/kPa

60 W100Y0
W95Y5
W90Y10
W85Y15
40
W80Y20
W75Y25

20
0 10 20
Frequency/Hz

Viscous modulus G″

60
G''/kPa

40

W100Y0
W95Y5
W90Y10
20 W85Y15
W80Y20
W75Y25

0 10 20
Frequency/Hz

0.54 Figure 1. Cont.

Loss tangent tan δ
0.52

0.50

0.48

0.46
a
 W100Y0
W95Y5
W90Y10
20 W85Y15
W80Y20
W75Y25
Foods 2020, 9, 256 7 of 15
0 10 20
Frequency/Hz

0.54
Loss tangent tan δ
0.52

0.50

0.48

0.46

G''/kPa 0.44

0.42
W100Y0
0.40 W95Y5
W90Y10
0.38 W85Y15
W80Y20
0.36 W75Y25

0 10 20
Frequency/Hz

Figure Frequency
1. 1.
Figure sweep
Frequency data
sweep forfor
data wheat dough
wheat with
dough different
with yam
different contents.
yam contents.

0 0
3.2.Compared to other
Effect of Yam samples,
Flour on DoughStructure
the Secondary W100Y0ofhad Protein G and G”. Meanwhile, G and G”
the lowest
Gluten
increased as yam flour content increased. Dough W75Y25 showed higher G0 and G” in comparison
FTIR spectra
with dough with lowof level
glutensubstitution
were measured to clarify
of yam flour. the effect
These of yam
results flour onthat
indicated gluten conformation
substitution of
(Figure 2). The secondary structure of gluten protein includes α-helix, β-sheet,
yam flour significantly affected the dough structure and gluten network, and the influence β-turn, random coil,
became
andpronounced
more so on. Amide asIthe
band was often
amount used
of yam to identify
flour theSimilar
increased. secondary
resultsstructure of protein.inIn
were illustrated the amide
studies of
0
adding potato flour [27]. Furthermore, the increasing trend of G and G” was different from that of
Cao [30] who argued that replacement of potato pulp decreased G0 and G”. These results could be
explained by the complex effects of protein, mucilage, and polysaccharides on dough viscoelasticity.
During bread making, the rheological characteristics were dominated by the cross-linking of gluten
network. Therefore, adding Yam flour to the dough, the gluten network had a higher crosslinking
density. As shown in Figure 1, tan δ increased in the dough when the level of yam flour addition
increased, which implied that the cross-linking degree in gluten network was reduced. Balestra [31]
reported that the higher value of tan δ might be due to the lower quantity of cross-linking in gluten
network. Tanδ of the dough decreased within 0.1–0.5 Hz. However, at 0.5–20 Hz, tanδ continued to
increase as the frequency continued to increase, indicating that the structure of the yam-wheat mixed
flour dough at higher frequencies was unstable and easily damaged.
The rheometer and texture data analyzer agreed with the farinograph results. These results
revealed that the dough with yam flour substitution had a reduced capacity to keep its shape during
proofing. Dough with yam flour substitution also decreased gas retention during the fermentation
process, which affects bread porosity.

3.2. Effect of Yam Flour on the Secondary Structure of Gluten Protein

FTIR spectra of gluten were measured to clarify the effect of yam flour on gluten conformation
(Figure 2). The secondary structure of gluten protein includes α-helix, β-sheet, β-turn, random coil,
and so on. Amide I band was often used to identify the secondary structure of protein. In the amide I
band, the secondary structure corresponding to each characteristic peaks were: 1615–1637 cm−1 and
1682–1700 cm−1 were β-sheet characteristic peaks; 1637–1645 cm−1 was random coil characteristic
peak; 1646–1664 cm−1 was the characteristic peak of α-helix; and 1664–1681 cm−1 was the characteristic
peak of β-turn [32]. α-helix and β-sheet were relatively ordered protein secondary structures, while
β-turn and random coil were disordered structures. The stability of α-helix and β-sheet was relatively
high. Figure 3 was the characteristic curve of gluten protein after fitting in the band of 1600–1700 cm−1 .
The content of secondary structure of gluten protein was listed in Table 3.
of β-turn [32]. α-helix and β-sheet were relatively ordered protein secondary structures, while β-turn
1646–1664 cm−1 was the characteristic peak of α-helix; and 1664–1681 cm−1 was the characteristic peak
and random coil were disordered structures. The stability of α-helix and β-sheet was relatively high.
of β-turn [32]. α-helix and β-sheet were relatively ordered protein secondary structures, while β-turn
Figure 3 was the characteristic curve of gluten protein after fitting in the band of 1600–1700 cm−1. The
and random coil were disordered structures. The stability of α-helix and β-sheet was relatively high.
content of secondary structure of gluten protein was listed in Table 3.
Figure 3 was the characteristic curve of gluten protein after fitting in the band of 1600–1700 cm−1. The
Foods 9, 256
2020, of
content secondary structure of gluten protein was listed in Table 3. 8 of 15
1600~1700 amide I band

1600~1700 amide I band

W75Y25
W75Y25
W80Y20
W80Y20
W85Y15
W85Y15
W90Y10
W90Y10
W95Y5
W95Y5
W100Y0
W100Y0

400 800 1200 1600 2000 2400 2800 3200 3600 4000
-1
(Wave number/cm )
400 800 1200 1600 2000 2400 2800 3200 3600 4000
-1
(Wave number/cm )
Figure 2. Fourier Transform Infrared Spectroscopy (FTIR) spectra of gluten protein in dough with
Figure 2. Fourier Transform Infrared
differentSpectroscopy (FTIR)
amounts of added spectra
yam flour.of gluten protein in dough with
Figureamounts
different 2. Fourier
ofTransform
added yamInfrared
flour. Spectroscopy (FTIR) spectra of gluten protein in dough with
different amounts of added yam flour.

Figure 3. Cont.
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9 of 1515

Figure 3. Second-derivative spectra in amide I band of gluten proteins enriched with different levels of
yam flour.
Figure 3. Second-derivative spectra in amide I band of gluten proteins enriched with different levels
of yam flour.
Table 3. Secondary structure of gluten protein in wheat dough enriched with different amounts of
yam flour.
Table 3. Secondary structure of gluten protein in wheat dough enriched with different amounts of
yam flour. Relative Proportion (%)
Sample
α-Helix β-Sheet β-Turn
Relative Proportion (%) Random Coil
Sample
W100Y0 α-Helix
32.24 β-Sheet
41.02 β-Turn
6.61 Random Coil
15.13
W95Y5
W100Y0 30.98
32.24 28.75
41.02 24.97
6.61 15.30
15.13
W90Y10 18.67 32.34 27.61 21.48
W95Y5 30.98 28.75 24.97 15.30
W85Y15 31.75 41.98 10.96 15.31
W90Y10
W80Y20 18.67
38.18 32.34
31.93 27.61
13.57 21.48
16.32
W85Y15
W75Y25 31.75
31.82 41.98
40.88 10.96
11.49 15.31
15.81
W80Y20 38.18 31.93 13.57 16.32
W75Y25 31.82 40.88 11.49 15.81
It has been generally accepted that the most stable secondary structure was β-sheets. Similarly,
It has beenin
an increasing generally accepted
the α-helix wouldthat alsothe most
lead to astable
moresecondary structure[33].
ordered structure wasAccording
β-sheets. Similarly,
to Table 3,
anthe
increasing in the α-helix would also lead to a more ordered structure
proportion of α-helix and β-sheet in W100Y0 gluten protein was 73.26%, and the proportion [33]. According to Table 3,
the proportion of α-helix and β-sheet in W100Y0 gluten protein was 73.26%,
of random coil was 15.13%. The secondary structure of gluten protein was dominated by β-sheet, and the proportion of
random
which iscoil was 15.13%.
consistent withThe secondary
previous reportsstructure of gluten
[34]. After adding protein was dominated
yam flour, the percentage by β-sheet, which
of α-helix and
isβ-sheet
consistent with previous reports [34]. After adding yam flour, the percentage
in gluten was lower than that in W100Y0, while the proportion of random coil was higher of α-helix and β-sheet
inthan
gluten was lower
W100Y0. A similarthanregularity
that in W100Y0, while the by
was demonstrated proportion
Chen [12].ofThe random
decrease coilinwas higher thanof
the proportion
W100Y0.
ordered A similar regularity
structures was demonstrated
and the increase in the proportion by Chen [12]. Thestructures
of disordered decrease in in gluten
the proportion
protein were of
ordered structures and the increase in the proportion
consistent with the results of dough rheological properties. of disordered structures in gluten protein were
consistent with the results of dough rheological properties.
3.3. Effect of Yam Flour on Antioxidant Capacity of Bread
3.3. Effect of Yam Flour on Antioxidant Capacity of Bread
Total phenolics contents (TPC), polysaccharides content, total flavonoids content (TFC), allantoin
Totaland
content phenolics contents (TPC),
radical scavenging capability polysaccharides
(IC50 Value) of bread content,withtotal flavonoids
different content
substitution ratio(TFC),
of yam
allantoin
flour werecontent
showed and inradical
Table 4.scavenging
As showedcapability
in Table 4,(IC 50 Value)
TPC, of bread with
polysaccharides, TFCdifferent substitution
and allantoin of bread
ratio of yamwith
increased flour were showed
substitution ratioindependence.
Table 4. As showed in Table
These results were4, in
TPC,
good polysaccharides,
agreement withTFC and
previous
allantoin
work [4].ofYam bread
flourincreased
addition with substitution
remarkably affected ratio DPPH radicalThese
the dependence. scavengingresults were inofgood
capability bread.
agreement
As could bewithseen previous
from Tablework
4, the [4].
IC50Yamvalues flour additionand
of W100Y0 remarkably
W95Y5 were affected the DPPH
the highest, radical
indicating that
scavenging
their radicalcapability
scavenging of bread. As could
capability was betheseen fromJing
lowest. Tableand4,Kitts
the IC50
[35] values
reported of that
W100Y0 and W95Y5
Maillard reaction
were the highest,
products showed indicating that their
certain radical radical scavenging
scavenging capability.capability was the that
It is interesting lowest. Jing and Kitts
a 10%–25% yam [35]
flour
reported thatcaused
substitution Maillard reactionimprovement
remarkable products showed in DPPHcertain radical
scavenging scavenging
capability. capability.
The radical It is
scavenging
interesting
capability that a 10%–25%
of bread yam dependent
was strongly flour substitution caused
on addition remarkable
of yam flour. DPPHimprovement in DPPH
radical scavenging
scavenging capability.
capability increase trendThe
wasradical
consistent scavenging
with TPC,capability of bread
polysaccharides, TFC,wasand strongly
allantoin dependent on
content of bread.
addition of yam flour. DPPH radical scavenging capability increase trend was consistent with TPC,
polysaccharides, TFC, and allantoin content of bread.
Foods 2020, 9, 256 10 of 15

Foods 2020, 9, x FOR PEER REVIEW 10 of 15

Table 4. Total phenolics content (TPC), polysaccharides, total flavonoids content, allantoin content, and
Tablevalues
IC50 4. Total phenolics
of bread withcontent (TPC),
different levelspolysaccharides,
of yam flour. total flavonoids content, allantoin content,
and IC50 values of bread with different levels of yam flour.
Total Phenolics Polysaccharides Total Flavonoids Allantoin IC50 Value
Sample
Content
Total Phenolics (mg GE/g)Polysaccharides
Content (mg/g) Total Content (mg RE/g) Allantoin
Flavonoids Content (mg/g)
Content (mg/mL)
Sample IC50Value（mg/mL）
Content（mg GE/g) Content（mg/g) Content（mg RE/g) e （mg/g)
W100Y0 0.467 ± 0.002 d 21.18 ± 0.14 f 0.162 ± 0.004 0.05 ± 0.011f 124.14 ± 1.62aa
W100Y0 0.467 ± 0.002d 21.18 ± 0.14f e 0.162 ± 0.004e 0.05 ± 0.011f 124.14 ± 1.62
W95Y5 0.470 ± 0.002 cd 22.45 ± 0.06 0.268 ± 0.006 d 0.279 ± 0.006e 130.09 ± 1.44aa
W95Y5 0.470 ± 0.002cd cd
22.45 ± 0.06e d
0.268 ± 0.006d c 0.279 ± 0.006e d
130.09 ± 1.44b
W90Y10
W90Y10 0.471 ±
0.471 ± 0.002cd 0.002 25.04
25.04 ± 0.11d± 0.11 0.378 ± 0.008
0.378 ± 0.008c 0.459 ± 0.066
0.459 ± 0.066 d 117.36±±0.82
117.36 0.82b
b c b c c
W85Y15 0.480 ±0.480
W85Y15 0.003±b 0.003 ± 0.04±
27.6527.65 c 0.04 0.475
0.475 ± 0.003
± 0.003 b 0.781
0.781 ± 0.012
± 0.012 c 113.37±±0.47
113.37 0.47c
W80Y20 b b 0.525 ± 0.015 a b d
W80Y20 0.481
0.481 ± 0.002±b 0.002 29.67
29.67 ± 0.09 ±
b 0.09 0.525 ± 0.015 a 0.894 ±
0.894 ± 0.0040.004
b 102.98 ± 0.95
102.98 ± 0.95d

W75Y25 0.002±
W75Y25 0.491 ±0.491 a 0.002a a 0.07a
± 0.07±
31.5231.52 0.516
0.516 ± 0.009
± 0.009 a a 1.063
1.063 ± 0.001
± 0.001 a a
0.95d
101.36±±0.95
101.36 d

The values
The values(mean
(mean ± standard
± standard deviation)
deviation) followed
followed by theby theletter
same sameinletter in the
the same same
column column
were were not
not significantly
different (p > 0.05) as determined by Duncan’s multiple range test (n = 3).
significantly different (p > 0.05) as determined by Duncan's multiple range test (n = 3).

3.4. Digital Images

3.4. Digital Images of
of Dough
Dough and
and Gluten
Gluten
As
As shown
shown in in Figure
Figure 4,
4, SEM
SEM of of W100Y0
W100Y0 showed
showed aa typical
typical structure
structure of
of wheat
wheat dough
dough [27].
[27]. The
The SEM
SEM
image
image ofof dough W95Y5 was
dough W95Y5 was similar
similar to
to W100Y0,
W100Y0, the the network
network structure
structure of
of gluten
gluten was
was well-established,
well-established,
with
with continuity,
continuity, thethe small
small and
and large
large starch
starch granules
granules were
were arranged
arranged tightly
tightly and evenly, and
and evenly, and they
they were
were
embedded in the network structure of gluten. These results agreed with the
embedded in the network structure of gluten. These results agreed with the changes in rheologicalchanges in rheological
properties
properties ofof dough
dough characterized
characterized by by rheometer
rheometer and and texture
texture analyzer. With the
analyzer. With the increase
increase ofof yam flour
yam flour
substitution,
substitution, the continuity of gluten network structure became worse, and the ability to wrap the
the continuity of gluten network structure became worse, and the ability to wrap the
starch granules became
starch granules becamelower.
lower.WhenWhenthe theadded
added amount
amount was
was 10%10% (W90Y10),
(W90Y10), thethe gluten
gluten protein
protein of
of the
the
doughdough began
began to appear
to appear discontinuous,
discontinuous, and
and thethestarch
starchgranules
granuleswere
wereexposed
exposed outside
outside the
the gluten
gluten
network. When the
network. When the added
added amount
amount was was 15%
15% (W85Y15),
(W85Y15), itit could
could be
be seen
seen that
that thethe starch
starch granules
granules
obviously
obviously separated
separated from from the
the gluten
gluten matrix.
matrix. When
When the the addition
addition amount
amount was
was 20%
20% or 25% (W80Y20
or 25% (W80Y20 or or
W75Y25), the gluten protein content decreased, and the gluten matrix was almost
W75Y25), the gluten protein content decreased, and the gluten matrix was almost invisible. The invisible. The number
of irregular
number voids increased,
of irregular and the gluten
voids increased, and thestructure was seriously
gluten structure damaged.damaged.
was seriously The scanning electron
The scanning
microscope results further
electron microscope confirmed
results further that substitution
confirmed of yam flour would
that substitution of yamaffectflourthewould
development
affect theof
continuous gluten network structure, destroyed the gluten network structure,
development of continuous gluten network structure, destroyed the gluten network structure, and and caused changes
in dough
caused characteristics.
changes in doughThese trends wereThese
characteristics. corresponding
trends were to previous results to
corresponding of reduction in dough
previous results of
stability
reductionand viscoelasticity
in dough stabilityof theviscoelasticity
and dough with yam of theflour addition.
dough Liu flour
with yam et al. addition.
[27] and Cao Liu et al. [30]
[27]
reported
and Cao that
et al.the substitution
[30] of potato
reported that flour and potato
the substitution pulp could
of potato breakpotato
flour and the gluten
pulpnetwork,
could break thereby
the
changing the viscoelasticity
gluten network, of dough.
thereby changing the viscoelasticity of dough.

W100Y0 W95Y5 WW95Y5

W10 SL
W90 G G
SS
SG
SS
G

W85Y15 G W80Y20 W75Y25

SL SL SS
SL
G
SS G
SS

Figure 4.4. Scanning

Scanningelectron
electron micrographs
micrographs of dough
of dough substituted
substituted with with different
different level oflevel
yam of yam
flour flour
(1000×).
(1000×).
SL: largeSL: large
starch starch granules,
granules, SS: smallSS: small
starch starch granules,
granules, and G:
and G: gluten gluten network.
network.

Figure 5 was a scanning electron microscope image of gluten at a magnification of 1000 times. It
can be seen from Figure 5 that W100Y0 showed a compact gluten structure with uniform pores, and
the edges of pores were smooth, indicating the formation of a solid gluten structure. When yam flour
Foods 2020, 9, 256 11 of 15

Figure 5 was a scanning electron microscope image of gluten at a magnification of 1000 times.
It can be 9,
Foods 2020, seen
x FORfrom
PEERFigure
REVIEW 5 that W100Y0 showed a compact gluten structure with uniform 11 pores,
of 15
and the edges of pores were smooth, indicating the formation of a solid gluten structure. When yam
was added
flour was addedat a at
low level
a low (W95Y5),
level (W95Y5),gluten protein
gluten protein could
couldstill
stillmaintain
maintainaagood
good structure.
structure. As
As the
proportion ofofyam
proportion yam flour
flour continued
continued to increase,
to increase, gluten structure
gluten structure changed significantly.
changed significantly. The smoothnessThe
smoothness
and uniformity andof uniformity
gluten were of gluten wereand
damaged, damaged,
the degreeandofthe degree increased
damage of damagewithincreased with the
the increase of
increase
ratio of ratio
of yam of yam
flour. flour.edges
The hole The hole
wereedges were smooth,
no longer no longerbut smooth, but were
were jagged, andjagged, and structure
the holes the holes
structure
began began to When
to collapse. collapse.
theWhen theofcontent
content of yam
yam flour flour25%
reached reached 25% (W75Y25),
(W75Y25), the glutenthe gluten
protein protein
structure
structure
was was honeycomb,
honeycomb, and the holesand the holesnoticeably
became became noticeably smaller, suggesting
smaller, suggesting a weakened a weakened gluten
gluten network
network structure.
structure. The changes The in
changes in the network
the network structurestructure
of glutenof gluten
may maytobe
be due thedue to the competition
competition between
between
the the in
mucilage mucilage
the yamin theand
flour yamtheflour
waterand the water
content content
of starch of starch
and protein, whichandredistributed
protein, whichthe
redistributed
moisture in thethe moisture
dough, causedin the dough,
partial caused partial
dehydration of thedehydration
gluten protein,of the glutenthe
changed protein,
glutenchanged
protein
the gluten
network protein and
structure, network
even structure, and collapse.
caused partial even caused partial
Several collapse.
reports Severalthat
have shown reports have of
the blend shown
bran
that
or thecould
salt blendhaveof bran or salt could
a significant have
impact onathe
significant impact on
gluten network the gluten network [12,36].
[12,36].

W100Y0 W95Y5 W90Y10

W85Y15 W80Y20 W75Y25

Figure 5.
Figure 5. SEM
SEM images
images of
of gluten
gluten substituted
substituted with
with different
different level
level of
of yam
yam flour.
flour. The
The magnifications
magnifications of
of
SEM images were 1000×.

3.5. Effect of
3.5. Effect of Yam
Yam Flour
Flour on
on Bread
Bread Baking
Baking Performance
Performance
The images of
The images of bread
bread crumb
crumbwere werepresented
presentedininFigure6.
Figure 6. It can
It can be be
seenseen
fromfrom Figure
Figure 6 that
6 that withwith
the
the increase of the yam flour addition ratio, the bread crumb showed more
increase of the yam flour addition ratio, the bread crumb showed more non-uniform and larger pores.non-uniform and larger
pores.
In orderIn to
order to further
further analyze analyze the structure
the structure of theofbread
the bread crumb,
crumb, a fractal
a fractal analysis
analysis waswas performed
performed on
on Figure 6, and the results were listed in Table 5. As the proportion of yam
Figure 6, and the results were listed in Table 5. As the proportion of yam flour increased, the fractal flour increased, the
fractal
dimensiondimension
of breadofincreased.
bread increased.
When yam Whenflouryamwasflour
addedwas
at added
5%, thereat 5%,
was there was no remarkable
no remarkable difference
difference in the fractal dimension of W95Y5 and W100Y0 (p
in the fractal dimension of W95Y5 and W100Y0 (p > 0.05). When the addition of yam > 0.05). When the addition of yam
flour reached
flour reached 10%, the fractal dimension was significantly higher than that
10%, the fractal dimension was significantly higher than that of W100Y0. When the addition of yam of W100Y0. When the
addition
flour wasof25% yam flour wasthe
(W75Y25), 25% (W75Y25),
fractal the fractal
dimension of the dimension of thewhich
bread was 1.80, breadwaswasan 1.80, whichofwas
increase an
5.26%
increase of 5.26% compared to W100Y0. The fractal dimension of the pore size
compared to W100Y0. The fractal dimension of the pore size distribution indicated the uniformity of distribution indicated
the
the uniformity of the pore size
pore size distribution. Thedistribution.
smaller the The smaller
fractal the fractalthe
dimension, dimension, the more
more uniform theuniform the
pore size
pore size distribution. On the other hand, the larger the fractal dimension,
distribution. On the other hand, the larger the fractal dimension, the more uneven the pore size the more uneven the pore
size distribution
distribution and and the the greater
greater thethe difference
difference between
between thepore
the poresizes.
sizes.TheTheincrease
increase of of the
the fractal
fractal
dimension indicates that the addition of yam flour changed the gas cell size formed
dimension indicates that the addition of yam flour changed the gas cell size formed inside the bread, inside the bread,
and the difference in gas cell size increased. This may be due to the addition
and the difference in gas cell size increased. This may be due to the addition of yam flour, which of yam flour, which
affects
affects the
the gas-holding
gas-holding properties
properties of of the
the dough. Crumb porosity
dough. Crumb porosity affects
affects the
the springiness
springiness and and hardness
hardness
of bread [37].
of bread [37].
It could be seen from Table 5 that as the ratio of yam flour increased, the hardness of bread
decreased first and then increased. When the amount of yam flour was more than 5%, the bread
hardness was higher than that of W100Y0, and it increased significantly as the ratio of yam flour
increased (p < 0.05). When the amount of yam flour was 25% (W75Y25), the bread hardness was 789.20
main reason was that the gluten content in the yam-wheat mixed flour decreased, the formation of
gluten network structure was affected and resulted in a reduction in the sensory quality of the bread.
Eke-Ejiofor J. et al. [8] reported that substitution of three leaf yam flour of no more than 5% would
produce bread without affecting the bread quality, while other researches showed that the yam flour
addition
Foods of256
2020, 9, no more than 20% had not significant negative impact on bread quality [6,11]. This
12may
of 15
be related to the differences in the composition of yam of different varieties.

W100Y0 W95Y5 W90Y10

W85Y15 W80Y20 W75Y25

Figure
Figure 6.
6. The
The effect of added yam flour on the characteristics of bread.

Table 5. Effects of yam flour addition on texture properties of bread.

Fractal Specific Overall

Sample Hardness (g) Springiness Cohesiveness Chewiness (g)
Dimension Volume (mL/g) Acceptability
W100Y0 1.71 ± 0.01c 472.32 ± 29.31de 0.639 ± 0.035a 0.567 ± 0.008a 170.90 ± 10.09b 4.47 ± 0.02a 98.0 ± 0.67a
W95Y5 1.72 ± 0.01c 422.87 ± 15.41e 0.618 ± 0.012ab 0.541 ± 0.019b 141.53 ± 12.37c 4.26 ± 0.06b 96.7 ± 1.64a
W90Y10 1.74 ± 0.01b 486.52 ± 23.30cd 0.590 ± 0.020bc 0.521 ± 0.010bc 149.41 ± 9.75c 4.06 ± 0.04c 92.2 ± 1.26b
W85Y15 1.75 ± 0.00b 530.83 ± 33.63bc 0.580 ± 0.005c 0.505 ± 0.011c 155.38 ± 10.24bc 3.97 ± 0.01d 89.1 ± 1.84c
W80Y20 1.76 ± 0.00b 575.94 ± 20.40b 0.582 ± 0.015c 0.510 ± 0.019c 171.22 ± 13.18b 3.89 ± 0.06d 80.8 ± 3.38d
W75Y25 1.80 ± 0.01a 789.20 ± 49.57a 0.586 ± 0.004bc 0.476 ± 0.010d 220.04 ± 12.24a 3.80 ± 0.07e 76.1 ± 2.23e
The values (mean ± standard deviation) followed by the same letter in the same column were not significantly
different (p > 0.05) as determined by Duncan’s multiple range test (n = 3).

It could be seen from Table 5 that as the ratio of yam flour increased, the hardness of bread
decreased first and then increased. When the amount of yam flour was more than 5%, the bread
hardness was higher than that of W100Y0, and it increased significantly as the ratio of yam flour
increased (p < 0.05). When the amount of yam flour was 25% (W75Y25), the bread hardness was
789.20 g, which was an increase of 67.10% compared with W100Y0. The increase in bread hardness was
consistent with the change in dough hardness. Hardness had a greater effect on the overall score of
bread, and had a negative correlation with bread quality. The smaller the hardness, the softer the bread.
Springiness decreased remarkably with the increase in the amount of yam flour (p < 0.05). There was a
positive correlation between the springiness and the quality of the bread. The larger the springiness,
the better the quality. The decrease in bread springiness may be caused by the increased proportion of
yam flour, the decreased gluten content of the mixed flour, the fact that the gluten network structure
was destroyed, and the gas retention of dough was reduced. Good air holding resulted in decreased
bread hardness and improved springiness. The tendency of bread cohesiveness and chewiness was
consistent with hardness.
The results of the effect of yam flour addition on specific volume were presented in Table 5.
The specific volume was remarkably (p < 0.05) affected by yam flour addition, the highest specific
volume was observed in W100Y0 and the lowest was in W75Y25. As Yam flour content increased, the
specific volume decreased. This showed that gas holding capacity of bread had decreased during
proofing and baking. This decrease would be due to destruction of gluten by adding yam flour, in
agreement with the change of gluten network. A similar trend was observed by addition of other
flours such as acha and bambara nut sourdough flour [38], bran [39,40], cassava, and three leaf yam [8].
Foods 2020, 9, 256 13 of 15

As can be seen from Table 5, the overall acceptability of bread W100Y0 was similar to W95Y5
(p > 0.05). When the amount of yam flour was more than 5%, there was a remarkable difference between
the overall acceptability of bread and bread W100Y0 (p < 0.05), and it decreased gradually when the
substitution of yam flour increased (p < 0.05). When the amount of yam flour was not more than
15%, the antioxidant activity of the bread was high and the degree of acceptance was relatively high.
The main reason was that the gluten content in the yam-wheat mixed flour decreased, the formation of
gluten network structure was affected and resulted in a reduction in the sensory quality of the bread.
Eke-Ejiofor J. et al. [8] reported that substitution of three leaf yam flour of no more than 5% would
produce bread without affecting the bread quality, while other researches showed that the yam flour
addition of no more than 20% had not significant negative impact on bread quality [6,11]. This may be
related to the differences in the composition of yam of different varieties.

4. Conclusions
Replacement of wheat flour with yam flour remarkably affected the dough characteristics and
bread qualities. With the increased percentage of yam flour, the breakdown, water absorption, degree
of softening, G0 and G” increased, while the peak time, viscosity value, setback, developing time,
stability, and extensibility decreased. Moreover, as the proportion of yam flour was increased, the
hardness, springiness and chewiness of dough decreased first and then increased. The microstructure
of dough and gluten was destroyed, and the proportion of a relatively stable structure in protein
secondary structure decreased by the addition of yam flour. In addition, the specific volume, hardness,
and springiness of bread were all affected by changes to dough rheological and structural properties,
following the addition of yam flour. The content of polyphenols, flavonoids, polysaccharides, allantoin,
and the scavenging ability of DPPH free radicals in bread showed an increase which depended on the
amount of yam flour. Compared with W100Y0, the addition of 5% yam flour didn’t exert significant
negative impact on overall acceptability of bread. Acceptable bread was made by the substitution of no
more than 15% yam flour for wheat flour in bread recipe. Adding Guihuai number 2 yam flour to bread
recipe will improve the nutritional value and antioxidant capability of bread. The results of the present
study demonstrated that Guihuai number 2 had broad application prospects in baked products.

Author Contributions: Conceptualization, Q.-M.L., X.-J.S., and Y.L.; validation, J.-H.Z.; formal analysis, P.Y.,
S.-Y.G., and F.W.; funding acquisition, Xiao-Jun Su; writing—original draft preparation, Q.-M.L. and Y.L.;
writing—review and editing, Q.-M.L. and X.-J.S. All authors have read and agreed to the published version of
the manuscript.
Funding: This research was funded by Scientific Research Fund of Hunan Provincial Education Department,
grant number16A100.
Conflicts of Interest: The authors declare no conflict of interest.

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