LWT - Food Science and Technology 174 (2023) 114438

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LWT
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Effects of mixed fermentation of different lactic acid bacteria and yeast on

phytic acid degradation and flavor compounds in sourdough
Luping Fang a, Weijun Wang b, Zhixia Dou a, Jie Chen a, Yuecheng Meng a, Liqin Cai a,
Yanhua Li a, *
a
College of Food & Biology Engineering, Zhejiang Gongshang University, Key Laboratory for Food Microbial Technology of Zhejiang Province, Hangzhou, 310018, China
b
Research & Development Institute, Nanjing Weigang Dairy Co., Ltd., Nanjing, 211100, China

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

Keywords: Sourdough plays an important role in improving product quality. In this study, single-strain and mixed
Sourdough fermentation of Lactobacillus paracasei LG0260, Lactobacillus plantarum LG1034, Lactococcus lactis LG0827,
Mixed fermentation Saccharomyces cerevisiae J2815 and Saccharomyces cerevisiae J8202 were used to prepare type II sourdough,
Phytic acid
which was used to evaluate the ability of mixed fermentation to improve the quality of sourdough and increase
Antioxidant activity
volatile flavor compounds. In sourdough, mixed fermentation reduced the number of lactic acid bacteria (LAB)
Volatile flavor compounds
and yeasts colonies, accompanied by a decrease in acidity. However, the organic acid analysis found that mixed
fermentation increased the content of acetic acid and reduced the content of lactic acid. Mixed fermentation
enhanced the degradation of phytic acid. It was found that sourdough MY1L3 degrades up to 96.6% of phytic
acid. In addition, the total polyphenol content of mixed fermentation was decreased, but the DPPH free radical
scavenging ability was enhanced. It was found by HS-SPME-GC-MS that mixed-fermented sourdough had more
complex flavor compounds, such as acids, aldehydes, and esters. The differences in the metabolic properties of
different starter cultures in sourdough will provide a theoretical reference for the improvement of sourdough
quality.

1. Introduction
Sourdough is the product of mixing wheat flour and water and fer­
mented by microorganisms such as lactic acid bacteria (LAB) and yeasts.
Its addition to bakery products could lead to products with higher
nutritional value, good organoleptic properties, and considerable tech­
nical properties (Gobbetti, Rizzello, Cagno, & Angelis, 2014), such as
increase free amino acid content (Curiel, Coda, Centomani, Summo, &
Rizzello, 2015), improve dough rheological properties (Pei et al., 2020)
and bread flavor (Xu et al., 2020), extend shelf life (Sun et al., 2020),
increase bioactive compounds (Gobbetti et al., 2018), etc. So it is very
popular.
Minerals such as potassium, phosphorus, magnesium, zinc, etc. Are
present in grains (Arendt, Moroni, & Zannini, 2011). Also, phytic acid is
present. However, phytic acid chelates with these minerals to form
phytate made them insoluble (Buddrick, Jones, Cornell, & Small, 2014),
Phytic acid degradation allowed an increase in mineral, free amino acid
(FAA) and protein bioavailability (Gobbetti et al., 2018). The use of
sourdough fermentation could effectively reduce phytic acid, which was
due to the production of some phytase by microbial metabolism, and the
pH reduction caused by fermentation, which activates the endogenous
enzyme system of the grain, thereby degrading phytic acid. Sourdough
wheat bread was considered a low glycemic index (GI) food (Wolter,
Hager, Zannini, & Arendt, 2014), and sourdough fermentation could
reduce the GI value of bread (Demirkesenbicak, Arici, Yaman, Karasu, &
Sagdic, 2021). This was mainly related to the production of organic
acids, especially lactic acid, which reduced starch digestibility, while
acetic and propionic acids prolonged gastric emptying time (Novotni
et al., 2011). Both the phenolic content and DPPH free radical scav­
enging ability of bread prepared from sourdough were improved (Sidari,
Martorana, Zappia, Mincione, & Giuffrè, 2020). This was because LAB
could activate some enzymes in the process of fermenting grains,
including phenolic acid reductase, phenolic acid dehydrogenase and
glucosidase, etc. The release of these enzymes increased the total
phenolic content of the dough, thereby improving the antioxidant ca­
pacity of the dough. At present, most of the use of sourdough is LAB

* Corresponding author.
E-mail address: liyanhua607@163.com (Y. Li).

https://doi.org/10.1016/j.lwt.2023.114438
Received 27 June 2022; Received in revised form 31 December 2022; Accepted 5 January 2023
Available online 6 January 2023
0023-6438/© 2023 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/).
L. Fang et al.
sourdough combined with baker’s yeast fermentation to prepare bread
dough. For the research on sourdough, most of the sourdough is fer­
mented by LAB, and it is the traditional type of fermentation type I
sourdough. The research on the co-fermentation of industrial type II
sourdough with LAB and yeasts is relatively weak. Type II sourdough is a
liquid sourdough that has several advantages over Type I (Carnevali,
Ciati, Leporati, & Paese, 2007): (1) easier management and reproduc­
ibility during operative conditions; (2) easier control fermentation pa­
rameters and additions nutrients for conditioning the microbial
performances; (3) Type II sourdough products have better sensory and
nutritional properties.
The flavor compounds produced during sourdough fermentation can
be mainly divided into non-volatile organic acids (for example lactic
acid) and volatile compounds, which are the result of co-fermentation of
lactic acid bacteria and yeasts. Yeasts in sourdough could produce
ethanol, propanol and other isoalcohols. Heterotypic lactic acid bacteria
mainly produce some specific esters, alcohols and aldehydes. While,
homotypic lactic acid bacteria mainly produce diacetyl and carbonyl
compounds (Salim-ur-Rehman, Paterson, & Piggott, 2006). It was found
that the content of volatile flavor compounds of the dough fermented by
sourdough was higher than that of the chemically acidified dough and
the products produced by sourdough fermentation were generally more
popular with consumers (Debonne et al., 2020).
In this study, LAB and yeasts with excellent comprehensive perfor­
mance screened in the laboratory were used to ferment sourdough with
single strain and mixed LAB and yeasts. To study the metabolites on
phytic acid degradation and flavor compounds, it provides a theoretical
basis for the mining of sourdough starter and the improvement of
sourdough fermentation technology.
2. Material and methods
2.1. Strains and flour
Five strains including three bacteria of LAB (Lactobacillus paracasei
LG0260, Lactobacillus plantarum LG1034, Lactococcus lactis LG0827) and
two strains of yeasts (Saccharomyces cerevisiae J2815, Saccharomyces
cerevisiae J2808) were isolated from the traditional fermented food in
our laboratory. The strain was stored in sterile 25% glycerol at − 80 ◦ C
before use. The flour was medium gluten bread flour produced by China
Oil Foodstuffs Corporation Company (Beijing, China).
2.2. Preparation of sourdough
LAB (10 mL/dL) were inoculated into MRS broth at 37 ◦ C for 12 h,
and yeasts (10 mL/dL) were inoculated into YPD liquid medium at 28 ◦ C
for 12 h, and were passaged twice. The bacterial suspension was
centrifuged (5000 g for 10 min at 4 ◦ C) to remove the supernatant, and
the bacteria were obtained after washing twice with sterile normal sa­
line. The bacterial cells were added to 100 g of UV-sterilized flour, and
125 g of sterile water at 30 ◦ C was added. Stirred and mixed the dough
evenly (DY value was 225, LAB content was 107 CFU/g, yeasts content
was 107 CFU/g). The number of sourdough involved in this study and
the composition of strains used for fermentation were shown in Table 1.
2.3. Determination of basic indexes of sourdough
The determination of basic physical and chemical indexes of sour­
dough according to the method of Liu et al. (2016). The different
fermentation time sourdough (10 g) was mixed with 90 mL aseptic sa­
line. The mixture was diluted to a suitable gradient by 10-fold gradient,
and 100 μL of different gradient dilutions were taken. The LAB were
counted in MRS agar by pouring method, and the yeast was counted by
plate coating method. They were placed in a constant temperature
incubator at 37 ◦ C and 28 ◦ C for 48 h, and the total number of microbial
colonies was counted in colony-forming units, expressed as CFU/g.
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LWT 174 (2023) 114438
Table 1
The number of sourdough and the composition of strains used for fermentation.
Sourdough number Strains used for fermentation
Y1 S.cerevisiae J2815
Y2 S.cerevisiae J2808
L1 L.paracasei LG0260
L2 L. plantarum LG1034
L3 Lactococcus lactis LG0827
MY1L1 S.cerevisiae J2815 and L.paracasei LG0260
MY1L2 S.cerevisiae J2815 and L. plantarum LG1034
MY1L3 S.cerevisiae J2815 and Lactococcus lactis LG0827
MY2L1 S.cerevisiae J2808 and L.paracasei LG0260
MY2L2 S.cerevisiae J2808 and L. plantarum LG1034
MY2L3 S.cerevisiae J2808 and Lactococcus lactis LG0827
The different fermentation time sourdough (10 g) was mixed with 90
mL distilled water. After fully mixing, the pH value of the mixture was
determined by pH instrument. The mixed solution was titrated with 0.1
mol/L NaOH standard solution to make pH reach 8.6. The total titration
acidity (TTA) of the sourdough was expressed in terms of consumption
volume.
2.4. Determination of organic acids
According to the method of Lefebvre, Gabriel, Vayssier, and Fon­
tagné-Faucher (2002) the high performance liquid chromatography
technology was used to determine the lactic acid and acetic acid in the
fermented sourdough extract. The sourdough sample (10 g) fermented
for 24 h was homogenized in 90 mL ultra-pure water at a speed of 12,
000 g and homogenized for 1 min. Homogenized solution (10 mL) was
mixed with 5 mL 0.5 mol/L H2SO4 and centrifuged at 8000 g for 10 min.
The supernatant was neutralized to pH 4.5 with 2 mol/L KOH, and the
volume was fixed to 25 mL with ultra-pure water and precipitated in ice
bath. The sample and standard solution were filtered with a 0.45 μm
filter membrane and then injected 10 μL into an Agilent 1260 Infinity
system (Boblingen, Germany). Chromatographic separation was ach­
ieved with a ZORBAX Eclipse XDB-C18 column (3.0 mm × 100 mm, 3
μm). The elution condition was that the volume ratio of 0.1 mol/L
K2HPO4 and methanol was 97:3 and the sample was eluted for 15 min.
The column temperature was 30 ◦ C. The detection wavelength was 210
nm, and the flow rate was 0.8 mL/min. The standard curve was con­
structed with organic acids standard solution (0.25–1.5 mg/mL).
2.5. Change of reducing sugar content in sourdough
Sourdough samples (5 g) with different fermentation time were
added with 45 mL distilled water, stirred at 50 ◦ C for 30 min to fully stir
the solution, and centrifuged at 5000 g for 5 min. The extract superna­
tant was added 1 mL DNS (3,5-dinitrosalicylic acid) and heated in a
boiling water bath for 5 min, and then the mixture was cooled to room
temperature with tap water and kept in the dark. The absorbance at 540
nm was measured with an automatic microplate reader. The standard
curve was made using glucose as reference.
2.6. Changes of phytic acid content in different sourdough
According to the method of Buddrick et al. (2014), the phytic acid
content of sour dough was determined. Sourdough sample (0.5 g) was
mixed with 30 mL of 0.5 mol/L HCl solution, and then extracted with
shaking at 150 g for 3 h. Then the mixture was centrifuged at 5000 g for
30 min to obtain supernatant. The supernatant (1 mL) was mixed with 2
mL of NH4Fe(SO4)2 solution (0.2 g of NH4Fe (SO4)2⋅12H2O was dis­
solved in 100 mL of 2 mol/L HCl solution, and dissolved in distilled
water to 1000 mL). The mixture was bathed in boiling water for 30 min,
cooled rapidly in ice bath and centrifuged at 6000 g for 30 min. The
supernatant (2 mL) was mixed with 3 mL of 1% bipyridine (1.0 g of 2,
L. Fang et al.
2′ -bipyridine and 1.0 mL of thioglycolic acid were dissolved in distilled
water, then the volume was adjusted to 1000 mL), and the absorbance
was measured at 519 nm. The standard curve was constructed with
phytic acids standard solution (0.00–0.20 mg/L).
2.7. Determination of total polyphenol and antioxidant activity
According to the method of Nionelli et al. (2014), the content of
soluble total polyphenol and antioxidant activity were determined. The
ethanol extract of 100 g/kg sourdough was prepared. The extract (0.5
mL) was placed in a 10 mL volumetric flask, followed by the addition of
1 mL Folin - phenol chromogenic agent, 2 mL 15% Na2CO3 solution, and
fixed volume to 10 mL with ultrapure water. The mixture was reacted at
room temperature for 2 h, and the absorbance value was determined at
765 nm. The standard curve was constructed with gallic acid standard
solution. The mixture of water and reagent was used as blank. The re­
sults were expressed as gallic acid equivalent (GAE) per g of sample.
The extract (0.1 mL) was mixed with 3.9 mL 25 mg/L DPPH ethanol
solution. The sample was reacted for 30 min under the condition of
avoiding light, and the absorbance was determined under 517 nm. The
mixture of 15% ethanol solution and reagent was used as blank. The
standard curve was constructed with vitamin E standard solution. The
results were expressed as vitamin E equivalent (VEE) per g of sample.
The extract (0.1 mL) was mixed with 3.9 mL ABTS+ working solution
(2.45 mmol/L K2(SO4)2 solution mixed with 7 mmol/L ABTS+ solution
in equal volume, reacted in dark for 12–16 h. And diluted to absorbance
of 0.7 ± 0.02 at 734 nm). It was placed under the condition of avoiding
light for 10 min, and the absorbance was determined under 517 nm. The
mixture of 15% ethanol solution and reagent was used as blank. The
standard curve was constructed with vitamin E standard solution. The
results were expressed as vitamin E equivalent (VEE) per g of sample.
2.8. Determination of volatile compounds
Volatile compounds in unfermented dough and fermented sour
dough were quantified by headspace solid-phase microextraction
coupled with gas chromatography-mass spectrometry (HS-SPME-
GCMS). In short, a mixture of 5 g sour dough freeze-dried powder, 10 mL
ultra-pure water and 100 μL 100 mg/L 4-methyl-2-pentanol (internal
standard) was filled into 20 mL screw vials fitted with
polytetrafluorethylene-silicone septa, reaction at 50 ◦ C for 30 min. The
volatiles were then sampled with an extraction fiber (UPELCO 50/30 μm
DVB/CAR/PDMS) into the headspace bottle for 30 min. After extraction,
the fiber was immediately inserted into the injection port of the GC-MS
for the analysis step at 250 ◦ C for 2 min. GC-MS analyses was carried out
on an Agilent 7890AGC 5975C system coupled with a quadrupole mass
filter for mass spectrometric detection (Agilent Technologies, Santa
Clara, CA). Chromatographic separation was carried out on the DB-WAX
capillary column (60.0 m × 0.25 mm × 0.5 μm, Agilent, USA). The
analysis parameters were as follows: the inlet temperature was 260 ◦ C.
The column flow rate was 1 mL/min. The column temperature was held
at 35 ◦ C for 3 min, then 2 ◦ C/min to 160 ◦ C, then 3 ◦ C/min to 200 ◦ C, and
finally 4 ◦ C/min to 240 ◦ C, held for 5 min. The temperature of the EI ion
source was 230 ◦ C, the temperature of the quadrupole was 150 ◦ C, and
the mass scanning range was 33–500 amu. The compounds were
tentatively identified by matching the mass spectra with the NIST14
mass spectral database.
2.9. Data analysis
All of the experiments were repeated at least three times. Means and
standard deviations were calculated. The results were analyzed by SPSS
version 26 software for Windows (SPSS Inc., USA). Duncan tests at a
significance level of P < 0.05 were performed for significance analysis.
Data analysis using Origin (Origin 2018, USA).
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LWT 174 (2023) 114438
3. Results and discussion
3.1. Changes of basic physical and chemical indicators during
fermentation
The acidification of LAB is the basis for the functional properties of
sourdough (Arendt, Ryan, & Bello, 2007), and it is of great significance
to study the acidification process of the system. In the early stage of
fermentation (0–4 h), the growth of LAB was slow, and it was in the
growth delay period, while the growth delay period of yeasts was not
obvious (Fig. 1A and B). Compared to the following stage, the rate of pH
decrease and the rate of TTA increase were both lower at this stage. After
4–8 h of fermentation, LAB and yeasts began to proliferate in large
quantities (Fig. 1C and D). At this stage, the growth and metabolism of
microorganisms were the most vigorous, and the rate of pH reduction
and the rate of TTA growth increased. From 8 to 12 h, Lactococcus lactis
LG0827 participated in the fermentation of three kinds of sourdough L3,
MY1L3, MY2L3, the growth rate of the colonies number of LAB
decreased significantly, and the pH of the sourdough decreased at a
slower rate. However, the LAB in other sourdoughs still maintained a
high growth rate. After 12 h, the colonies number of LAB in each sour­
dough gradually became flat, which was a relatively stable state. How­
ever, the colonies number of yeasts showed different trends, and the
colonies number of yeasts in sourdough Y1 and Y2 still maintained an
increasing trend, while the colonies number of yeasts in the mixed
fermentation group showed a decreasing trend. When fermented for 24
h, the counts of LAB and yeasts in different sourdoughs were signifi­
cantly different. The counts of LAB ranged from 8.39 to 9.00 lgCFU/g,
and the counts of yeasts ranged from 7.20 to 8.22 lgCFU/g. At the same
time, it was found that the colonies number of LAB and yeasts in the
mixed fermentation group decreased compared with the single-strain
sourdough fermented by the corresponding strain, indicating that
when yeasts and LAB coexist, they have a certain antagonistic effect on
their growth. This was also reflected by the corresponding pH and TTA,
the co-fermented sourdough had a higher pH and a lower TTA. However,
Xu et al. (2019) found that the co-culture of Lactobacillus sanfrancisco
and yeasts had no effect on the colonies number of yeasts. But only on
the number of Lactobacillus sanfrancisco colonies, which might be caused
by the different interactions between different LAB and yeasts. The
growth of LAB in sourdough was negatively correlated with TTA.
The higher the colonies number of LAB, the higher the acidity, which
was also reflected in the study (Lancetti, Sciarini, Pérez, & Salvucci,
2020). When fermented for 24 h, the pH values of the same type of
sourdough (samples fermented by single yeasts, single LAB, and mixed
cultures respectively) were similar, but the TTA of mixed fermented
sourdough showed significant difference. The results probably due to
the differences in organic acid metabolism of different LAB, the titration
jump ranges of different organic acids were different, and the lye
consumed during titration was also different (Sahin et al., 2018). It was
found that among all the sourdoughs, the colonies number of LAB and
TTA in the sourdough fermented by Lactococcus lactis LG0827 were
smaller than that of other LAB. This might come from Lactococcus lactis
LG0827 belonged to homofermentative LAB, and it was mainly for lactic
acid metabolism. While L. paracasei LG0260 and L. plantarum LG1034
were heterofermentative LAB, their metabolites were mainly for acetic
acid metabolism, but also little acetic acid. Moreover, in the study of
others, it was also found that the homofermentative type of LAB had
weak acidifying ability in sourdough, and the number of viable bacteria
was less (Lancetti, Sciarini, Pérez, & Salvucci, 2021). Lactobacillus
plantarum LG1034 in this study had relatively strong acidifying ability,
which was similar to the results of some studies with better acidifying
ability of Lactobacillus plantarum strains (Liu et al., 2020; Vuyst et al.,
2014). This seemed to be related to the larger genome of Lactobacillus
plantarum, which made it suitable for the sourdough system. Although
TTA was reduced when LAB and yeasts coexist, co-fermentation might
increase the types of organic acids and more abundant volatile flavor
L. Fang et al. LWT 174 (2023) 114438

Fig. 1. Changes in basic indicators of sourdough.

Lactic acid bacteria (LAB) viable counts (A), yeasts
viable counts (B), pH (C) and total titration acidity
(D), respectively. Capital letters refer to different
samples: fermented sourdough inoculated S. cerevisiae
J2815 (Y1), S. cerevisiae J2808 (Y2), L. paracasei
LG0260 (L1), L. plantarum LG1034 (L2), Lactococcus
lactis LG0827 (L3), S. cerevisiae J2815 and L. paracasei
LG0260 (MY1L1), S. cerevisiae J2815 and L. plantarum
LG1034 (MY1L2), S. cerevisiae J2815 and Lactococcus
lactis LG0827 (MY1L3), S. cerevisiae J2808 and
L. paracasei LG0260 (MY2L1), S. cerevisiae J2808 and
L. plantarum LG1034 (MY2L2), S. cerevisiae J2808 and
Lactococcus lactis LG0827 (MY2L3).

compounds, especially esters (Xu et al., 2019), which was also reflected
in the subsequent results of volatile flavor compounds.

3.2. Organic acid content of different sourdoughs

Organic acids play an important role in sourdough systems, affecting

product structure, organoleptic properties and shelf life (Settanni et al.,
2013). Fig. 2 shows lactic acid content, acetic acid content and the
quotient of fermentation (QF), QF was determined as the ratio of lactic
acid content to acetic acid content at the end of fermentation of
sourdough.
Yeast-fermented dough accumulated a certain amount of acetic acid,
but no lactic acid production was detected. Lactobacillus-fermented
sourdough accumulated a large amount of lactic acid, while the in­
crease in acetic acid content was much smaller. Among them, the
sourdough L1 with the highest lactic acid content reached 4.35 mg/g,
and the mixed sourdough MY1L1 with the highest acetic acid content
was 1.24 mg/g. Comparing the sourdough fermented with the single
strain and the mixed strain, the study found that with the addition of
yeasts, the lactic acid content decreased. This was due to the reduction
in the colonies number of LAB, which in turn reduced the production of
Fig. 2. Lactic acid content, acetic acid content and the quotient of fermentation
lactic acid. However, the acetic acid content of mixed-fermented sour­ (QF) value of sourdough fermented for 24 h. QF was the ratio of lactic acid
dough was higher than the sum of the corresponding lactobacillus- content to acetic acid content at the end of fermentation of sourdough. CT0
fermented or yeast-fermented sourdough, which might be due to the means unfermented dough. Capital letters refer to different samples: fermented
yeasts metabolizing the glucose produced by LAB (Nistal et al., 2012), sourdough inoculated S. cerevisiae J2815 (Y1), S. cerevisiae J2808 (Y2),
and increased the production of acetic acid. The acetic acid content of L. paracasei LG0260 (L1), L. plantarum LG1034 (L2), Lactococcus lactis LG0827
sourdough L3 was smaller than that of L1 and L2, and the difference was (L3), S. cerevisiae J2815 and L. paracasei LG0260 (MY1L1), S. cerevisiae J2815
significant (P < 0.05). This was mainly due to the different fermentation and L. plantarum LG1034 (MY1L2), S. cerevisiae J2815 and Lactococcus lactis
pathways of Lactococcus lactis LG0824, Lactobacillus plantarum LG1034 LG0827 (MY1L3), S. cerevisiae J2808 and L. paracasei LG0260 (MY2L1),
and Lactobacillus paracasei LG0260. The former belonged to homo­ S. cerevisiae J2808 and L. plantarum LG1034 (MY2L2), S. cerevisiae J2808 and
Lactococcus lactis LG0827 (MY2L3).
fermentation, while the latter two belonged to heterofermentation. The
less acetic acid in L3 may be due to the production of microorganisms
carried by the flour itself. QF in the range of 1.5–4 was considered strains of LAB and yeasts were co-fermented.
positive in sourdough bread making (Gobbetti, De Angelis, Corsetti, & Di
Cagno, 2005), the QF values of the 3 groups of lactobacillus fermented 3.3. Variation of reducing sugar content in sourdough
sourdough in this study were higher than others, but it was found that
the QF value can be reduced to a more satisfactory range when the Starch is decomposed into reducing sugar under the action of

4
L. Fang et al.
amylase, and the reducing sugar is decomposed into monosaccharide,
which is the main raw material for yeasts respiration. Fig. 3 shows the
variation of reducing sugar content in different sourdoughs. The content
of reducing sugar in sourdough fermented by LAB showed an overall
upward trend with the increase of fermentation time, while the content
of reducing sugar in yeasts fermented dough showed a trend of first
increasing and then decreasing. Because in the early stage of fermen­
tation, starch was hydrolyzed into reducing sugar under the action of
various endogenous amylases of microorganisms, while yeasts respira­
tion could consume a part of reducing sugar. When the consumption rate
of reducing sugar exceeded the production rate, the reducing sugar
content began to decrease. The reducing sugar content of L3 continued
to increase. At 24 h, compared with other sourdoughs, the reducing
sugar content was the highest, which was 31.89 mg/g, and the differ­
ence was significant (P < 0.05). A decrease in pH could increase the
activity of amylase and expand the starch molecules, which was bene­
ficial for amylase to function (Uthumporn, Zaidul, & Karim, 2010).
However, when the pH of sourdough was less than 4.0, the activity of
amylase was inhibited, and LAB could consume part of the mono­
saccharide (Lancetti et al., 2021). Since the pH of sourdough L3 was
higher than that of L1 and L2, the inhibition of amylase was smaller,
resulting in a continuous increase in the reducing sugar content, while
LAB could consume part of the disaccharide, resulting in a decrease in
the reducing sugar content of L2. The change of reducing sugar in mixed
fermented sourdough could also explain the relationship between the
above-mentioned yeasts and LAB for the formation and utilization of
reducing sugar. The reducing sugar content of sourdough MY1L3 was
significantly different from other mixed sourdough (P < 0.05), reaching
20.99 mg/g. Under the action of LAB, sourdough produced a large
amount of reducing sugar for yeasts to use, which may explain the
reason why the previous mixed fermentation increased the acetic acid
content.
3.4. Phytic acid degradation in sourdough
Fig. 4 shows the change of phytic acid content in the fermentation
process of different sourdoughs. Compared to the phytic acid content of
the unfermented sourdough (1.39 mg/g), the phytic acid content of all
sourdoughs decreased gradually over time. When fermented for 24 h,
the content of phytic acid was in the range of (0.05–0.79 mg/g), which
decreased by 42.9%–96.6%. It shows that after adding phytase-
producing strains, fermentation can effectively degrade phytic acid,
and the fermentation time is an important factor affecting the degra­
dation of phytic acid, this has also been demonstrated in other study
(Buddrick et al., 2014). The phytic acid degradation rate of single-strain
fermented sourdough was lower than that of mixed fermented group,
which may be due to the combined effect of phytase produced by LAB
and yeasts to degrade phytic acid. The ability of yeast-fermented dough
to degrade phytic acid was not as good as that of LAB-fermented sour­
dough, partly because the phytase activity of yeasts in the selected strain
5
LWT 174 (2023) 114438
was lower than that of LAB. Another aspect was that the pH of LAB
sourdough was lower, which stimulated the endogenous phytase activity
of grains, resulting in its ability to degrade phytic acid was stronger than
that of yeasts sourdough. The phytic acid degradation rates of sour­
dough L3, MY1L3 and MY2L3 fermented by Lactococcus lactis LG0827
reached 92.9%, 96.6% and 92.5%, and the degradation effect was
particularly significant, which may be due to the higher phytase of the
selected Lactococcus lactis LG0827. Fekri, Torbati, Khosrowshahi,
Shamloo, and Azadmard-Damirchi (2020) believed that the reduction of
phytate was related to microbial phytase, and some researchers believed
that the degradation of phytate during fermentation was mainly due to
the action of endogenous phytase in grains. Compared with other study
(Montemurro et al., 2020), the sourdough in this study had a higher
phytic acid degradation rate, which may be due to less phytic acid in the
fermentation substrate, and the phytase activity of the selected
microorganism.
3.5. Total polyphenol content and antioxidant properties of different
sourdoughs
Phenolics are considered to be of great dietary importance due to
their antioxidant and anticancer properties (Călinoiu & Vodnar, 2018).
Table 2 shows the total polyphenol content and antioxidant activity of
unfermented dough CT0 and fermented 24 h sourdough. Compared with
CT0, the total polyphenol content, DPPH, and ABTS+ free radical
scavenging ability in each group of fermented dough increased signifi­
cantly (P < 0.05), but the ability of yeasts to enhance total polyphenol
content and antioxidant activity was not as good as that of LAB. Among
them, Lactobacillus plantarum LG1034 had the strongest ability to in­
crease the total polyphenol content of sourdough by 82.6%. The DPPH
free radical scavenging ability of L2 fermented by Lactobacillus planta­
rum LG1034 was the strongest, which was 3.41 times that of CT0. The
free radical scavenging ability of ABTS+ in sourdough L3 fermented by
Lactococcus lactis LG0827 was 11.96 times that of CT0. However,
co-fermentation improved the DPPH free radical scavenging ability, and
the levels of sourdough MY2L2 and MY1L2, which was 3.73 and 3.68
times that of CT0 respectively. Overall, Lactobacillus plantarum LG1034
had the best effect on improving the total polyphenol content and
antioxidant activity of sourdough.
From the above results, LAB played a leading role in the enhance­
ment of antioxidant properties of sourdough, and different microbial
fermentations produce different effects, which were caused by the
inactivation of active compounds as well as the activation of inactive
compounds (Sidari et al., 2020). It had shown that acidification can
improve the extractability of phenol, and LAB could increase the hy­
drolysis of complex phenols and the ability to scavenge free radicals
(Sidari et al., 2020). Filannino, Cagno, and Gobbetti (2018) found that
specific glycosyl hydrolases in Lactobacillus plantarum and Lactobacillus
rhamnosus decomposed the plant cell wall matrix, resulting in the release
of bound phenolic compounds, which facilitated the depolymerization
Fig. 3. Changes in reducing sugar content (RSC) in
different sourdough. RSC of single-strain fermented
dough (A). RSC of mixed-fermented dough (B). Cap­
ital letters refer to different samples: fermented
sourdough inoculated S. cerevisiae J2815 (Y1),
S. cerevisiae J2808 (Y2), L. paracasei LG0260 (L1),
L. plantarum LG1034 (L2), Lactococcus lactis LG0827
(L3), S. cerevisiae J2815 and L. paracasei LG0260
(MY1L1), S. cerevisiae J2815 and L. plantarum LG1034
(MY1L2), S. cerevisiae J2815 and Lactococcus lactis
LG0827 (MY1L3), S. cerevisiae J2808 and L. paracasei
LG0260 (MY2L1), S. cerevisiae J2808 and L. plantarum
LG1034 (MY2L2), S. cerevisiae J2808 and Lactococcus
lactis LG0827 (MY2L3).
L. Fang et al. LWT 174 (2023) 114438

Fig. 4. Changes in phytic acid content (PAC) in

different sourdough. PAC of single-strain fermented
dough (A). PAC of mixed-fermented dough (B). Cap­
ital letters refer to different samples: fermented
sourdough inoculated S. cerevisiae J2815 (Y1),
S. cerevisiae J2808 (Y2), L. paracasei LG0260 (L1),
L. plantarum LG1034 (L2), Lactococcus lactis LG0827
(L3), S. cerevisiae J2815 and L. paracasei LG0260
(MY1L1), S. cerevisiae J2815 and L. plantarum LG1034
(MY1L2), S. cerevisiae J2815 and Lactococcus lactis
LG0827 (MY1L3), S. cerevisiae J2808 and L. paracasei
LG0260 (MY2L1), S. cerevisiae J2808 and L. plantarum
LG1034 (MY2L2), S. cerevisiae J2808 and Lactococcus
lactis LG0827 (MY2L3).

antioxidant activity of ABTS+.

Table 2
Analysis of total polyphenol content and antioxidant activity of sourdough
during 24 h fermentation. 3.6. Volatile flavor compounds in different sourdoughs
Sample TPC GAE Multiple DPPH Multiple ABTS+ Multiple
(mg/g) VEE VEE In this study, the volatile flavor compounds of 11 kinds of different
(mg/g) (mg/g) sourdoughs, the commercial sourdough (LB) and the unfermented
CT0 0.420 ± – 0.126 ± – 0.050 ± – dough (CT0) were detected by HS-SPME-GC-MS and quantified by in­
0.004g 0.006e 0.011g ternal standard. The results are shown in Table 3. A total of 50 kinds of
Y1 0.521 ± 0.24 0.360 ± 1.85 0.487 ± 8.70
typical flavor compounds were detected, including 12 kinds of alcohols,
0.004f 0.087d 0.050ef
Y2 0.501 ± 0.19 0.368 ± 1.92 0.610 ± 11.17
12 kinds of acids, 12 kinds of esters, 3 kinds of ketones, 8 kinds of al­
0.001f 0.097d 0.055cd dehydes and 3 kinds of other compounds. Table 4 shows the statistical
L1 0.724 ± 0.72 0.519 ± 3.11 0.566 ± 10.28 results of the content of volatile flavor compounds in various categories.
0.050b 0.071abc 0.049de Compared with CT0, the process of fermentation significantly increased
MY1L1 0.704 ± 0.67 0.535 ± 3.24 0.664 ± 12.23
the types and contents of flavor compounds, indicating that fermenta­
0.003bc 0.031abc 0.069bc
MY2L1 0.689 ± 0.64 0.534 ± 3.23 0.723 ± 13.41 tion not only imparted the unique flavor to sourdough, but also helped
0.011bcd 0.034abc 0.069ab sourdough to have a stronger aroma. For the results of this study, the
L2 0.768 ± 0.83 0.557 ± 3.41 0.620 ± 11.36 types and contents of volatile flavor compounds produced by yeast-
0.025a 0.036ab 0.057cd fermented sourdough were much higher than those of LAB-fermented
MY1L2 0.721 ± 0.72 0.592 ± 3.69 0.804 ± 15.03
0.006b 0.030a 0.066a
sourdough, and the differences were mainly in the content of alcohols
MY2L2 0.698 ± 0.66 0.598 ± 3.73 0.608 ± 11.12 and esters. Mixed fermentation, compared with the single-strain fer­
0.010bc 0.057a 0.060cd mented dough, the types of volatile flavor compounds increased, but the
L3 0.681 ± 0.62 0.470 ± 2.72 0.650 ± 11.96 total content decreased, mainly because the content of alcohols
0.013cd 0.040c 0.072bcd
decreased, while the contents of acids, esters and aldehydes increased.
MY1L3 0.655 ± 0.56 0.482 ± 2.81 0.625 ± 11.47
0.009d 0.045bc 0.068cd This was consistent with the study that diverse fermenting microbial
MY2L3 0.603 ± 0.44 0.545 ± 3.32 0.642 ± 11.80 populations lead to diverse and intense food flavors (Xu et al., 2019).
0.001e 0.052abc 0.069bcd Among all samples, Y1 had the highest content of alcohols and ke­
The results of total polyphenol content (TPC) were expressed as gallic acid tones, reaching 1204.11 mg/kg and 2.23 mg/kg, respectively. The
equivalent (GAE) per g of sample. The results of DPPH and ABTS+ were highest content of volatile acids and aldehydes was found in MY1L2,
expressed as vitamin E equivalent (VEE) per g of sample. CT0 means unfer­ which was 60.11 mg/kg and 61.76 mg/kg, respectively. The highest
mented dough. The multiple indicates the fold increase compared to CT0. content of esters was found in LB, followed by MY1L2, containing
Different superscript lowercase letters indicate significance between columns (P 118.91 mg/kg and 101.93 mg/kg, respectively. It was also found that
< 0.05). the contribution of two yeast strains to the production of 3-methyl-1-
butanol, 3-methylthiopropanol and phenylethanol was undeniable,
of high molecular weight phenolic compounds (Katina et al., 2007). because these compounds were detected in all yeast-containing sour­
Bartkiene et al. (2017) reported that inoculation of Pediococcus lactis into doughs, but not in LAB sourdough. This result was similar to those of
barley sourdough increased phenolic content by 34% and free radical studies (Ravyts & Vuyst, 2011; Xu et al., 2020). This may be due to the
scavenging activity by 79%. The results of this study and other studies conversion of methionine and phenylalanine to 3-methylthiopropanol
had differences in value due to differences in research methods, and phenylethyl alcohol by S. cerevisiae through the Ehrlich pathway.
fermentation strains and substrates, but the change trend was consistent Moreover, the content of phenylethyl alcohol in the compound fer­
(Sripo, Phianmongkhol, & Wirjantoro, 2016). The three groups of mented sourdough increased, indicating that the mixed fermentation of
sourdough fermented by Lactococcus lactis LG0827 had lower total LAB and yeasts can promote the Ehrlich pathway of yeasts.
polyphenol content and DPPH free radical scavenging ability than those In this study, aldehydes were rarely found in yeasts single strain
fermented by the other two LAB, which were related to the colonies fermented dough, but high in sourdough fermented by LAB. The content
number of LAB and the degree of acidification in the sourdough, and of aldehydes in sourdough L1 and L2 was lower than that in L3. This may
these results were consistent with the correlations between total poly­ be due to the fact that LAB in L1 and L2 were heteromorphic fermented
phenol and antioxidant properties, bacterial load and pH found in other LAB. This type of LAB had higher ability to reduce aldehydes to other
study. However, the changes in the free radical scavenging ability of compounds (such as corresponding alcohols) (Kaseleht, Paalme, Mih­
ABTS+ were slightly different, which may be due to the different anti­ halevski, & Sarand, 2011), which was also reflected in the study of Liu
oxidant properties of different phenolics (Fardet, Rock, & Rémésy, 2008; et al. (2020), benzaldehyde and hexanal (grass-flavored) were the main
Sidari et al., 2020), so the low phenolic sourdough showed high aldehydes in compound sourdough, and the content of mixed

6
L. Fang et al. LWT 174 (2023) 114438

Table 3
The volatile compounds identified in different sourdough fermented for 24 h.
Volatile compounds Flavor compound content (mg/kg sourdough)

Y1 Y2 L1 L2 L3 MY1L1 MY1L2 MY1L3 MY2L1 MY2L2 MY2L3 LB CT0

alcohols
2-ethyl-1-hexanol 377.82 157.85 0.51 0.19 0.16 405.13 485.83 411.46 201.67 169.99 176.86 84.25 0.62
3-methylthiopropanol 10.90 6.58 nd nd 0.14 23.75 27.28 22.44 22.95 20.20 20.65 79.31 nd
phenylethyl alcohol 685.82 380.38 0.18 0.05 nd 504.86 492.19 531.36 265.35 242.24 302.09 69.50 nd
ethanol 68.11 74.88 1.16 nd 0.33 80.22 84.07 49.04 65.56 50.52 54.21 31.81 nd
isobutanol nd nd nd nd nd nd 5.00 4.75 6.58 5.68 nd 25.64 nd
3-methyl-1-butanol 23.63 25.85 18.48 12.59 18.80 26.54 25.69 22.80 23.30 19.77 25.38 23.91 4.29
1-butanol 3.22 2.23 17.44 14.62 30.65 24.32 35.80 30.80 19.44 22.50 20.62 11.28 1.06
n-heptanol 5.90 6.09 6.45 3.90 3.22 11.63 19.08 14.25 9.69 10.07 16.96 6.72 2.07
1-hexanol 4.21 1.37 nd nd nd 9.96 12.23 9.81 4.32 4.25 4.37 5.44 nd
1-nonanol 14.54 22.45 nd nd nd 21.24 25.77 24.07 14.80 17.95 18.71 5.13 nd
pentanol 39.13 15.16 nd nd nd 26.46 26.55 23.81 12.33 10.68 11.28 3.64 nd
1-octanol 3.14 4.70 0.47 0.36 0.35 8.15 8.39 7.55 8.09 8.07 8.30 3.61 nd
acids
isobutyric acid 2.93 2.92 0.71 0.58 0.89 3.33 3.70 3.13 2.42 2.32 2.72 3.38 0.22
DL-phenylsuccinic acid 5.87 3.81 1.14 1.38 1.65 13.35 15.21 13.56 10.95 6.62 8.96 3.38 0.33
benzoic acid 1.91 1.47 2.67 0.50 0.53 nd 2.49 2.92 nd 1.94 1.79 2.78 nd
acetic acid 1.88 2.22 0.71 0.48 0.72 2.85 3.19 2.69 2.04 2.05 2.24 2.11 0.15
heptanoic acid 4.38 5.14 5.58 4.68 4.53 17.92 19.22 15.79 15.24 15.14 12.16 2.06 1.63
hexanoic acid nd 1.11 2.29 1.87 2.24 1.98 2.33 2.14 1.98 1.73 1.83 1.96 0.81
valeric acid 1.96 nd nd nd 1.61 4.14 2.86 0.32 nd nd 1.39 1.78 nd
caprylic acid nd nd 2.00 1.36 2.86 3.12 5.68 4.53 2.65 2.55 2.25 1.48 nd
lauric acid nd nd nd nd nd 3.60 4.32 4.32 2.69 2.34 2.57 1.12 nd
butyric acid 9.11 22.19 nd nd nd 14.46 17.34 14.13 10.02 10.57 23.72 1.08 nd
decanoic acid 1.18 2.49 1.60 1.50 0.56 2.86 2.44 1.75 7.00 4.56 3.30 0.99 0.16
nonanoic acid nd nd nd nd nd 0.50 0.64 nd nd nd nd 0.57 nd
esters
ethyl propanoate nd nd nd nd 0.15 2.71 0.87 nd 0.38 0.38 0.34 0.55 nd
butyric acid ethyl ester 1.96 1.82 0.62 0.46 0.76 2.37 2.59 2.02 1.34 1.77 1.40 0.54 nd
ethyl heptanoate 1.19 1.09 nd nd nd 1.31 2.15 1.62 2.80 2.39 2.45 0.54 nd
ethyl caprate nd nd 0.28 0.17 0.36 0.77 1.15 0.85 0.71 0.49 0.51 0.36 nd
S-methyl thiohexanoate 0.77 0.93 nd nd nd nd 0.91 0.82 nd 0.37 0.46 0.35 nd
ethyl hexanoate 0.39 0.71 0.11 0.11 0.07 0.60 0.59 0.55 0.28 0.33 0.37 0.34 1.06
ethyl lactate nd nd nd nd nd 1.54 0.63 2.99 0.74 0.57 0.79 0.26 nd
etheyl octanoat 5.37 10.75 nd nd nd 4.11 7.27 6.02 3.44 3.74 4.76 0.26 nd
3-methoxy-3-methylbutyl acetate 1.44 3.13 0.21 0.13 0.13 1.53 1.46 1.00 0.58 0.55 1.50 0.22 13.24
ethyl acetate 1.31 3.09 nd nd nd 1.01 nd nd 0.98 nd nd nd nd
isobutyl acetate 0.57 1.28 nd nd nd 0.28 0.27 nd nd 0.28 0.35 nd 0.12
isoamyl acetate 1.00 1.02 0.17 0.12 0.28 0.77 1.14 0.76 0.68 0.60 0.66 nd 0.5
ketones
3-octen-2-one 1.83 0.96 nd nd nd 2.63 2.40 1.49 0.99 0.57 0.31 nd nd
2,3-pentanedione 1.11 0.95 nd nd nd nd nd nd nd nd nd nd 0.52
3-methyl-2-butanone 1.77 0.83 nd nd nd 0.45 nd 0.49 nd nd nd nd nd
aldehydes
(E)-2-octenal 1.12 0.79 nd nd nd nd nd nd nd nd nd nd nd
2-heptenal 0.64 0.50 nd nd nd 2.14 2.10 1.98 0.91 0.89 1.05 nd nd
benzaldehyde nd 0.29 nd nd nd 0.26 0.67 0.49 nd 0.29 1.13 nd nd
hexanal nd 0.21 0.33 0.26 0.18 0.80 1.01 0.96 0.37 0.40 0.84 nd nd
octadecanaldehyde nd nd nd nd nd nd nd nd nd nd nd nd nd
acetaldehyde nd nd 0.34 0.20 0.15 0.51 0.49 0.46 0.26 nd 0.52 nd nd
isovaleraldehyde nd nd 0.15 0.10 0.25 0.40 0.80 0.56 0.37 0.26 0.33 nd nd
pentanal nd nd nd 0.98 2.65 nd nd nd nd nd nd nd nd
others
2,2,4,6,6-pentamethylheptane nd nd 0.60 0.54 0.72 nd 1.12 1.01 nd nd 0.54 nd nd
2-pentylfuran nd nd 0.79 0.23 1.09 nd nd nd nd nd nd nd nd
acetoin nd nd 0.33 0.34 nd nd 0.27 nd 0.27 nd nd nd nd

The data in the table was the quantity calculated by the internal standard. Nd, not detected. LB was the commercial sourdough for comparison. CT0 means unfermented
dough.

fermentation increased significantly.
MY1L2 and MY2L3 contained 12 kinds of acidic volatile compounds.
Acetic acid, caproic acid and caprylic acid were the most important acids
in sourdough. The content of acetic acid in L1 and L2 was higher than
that in L3. The mixed fermentation produced more acetic acid, which
was consistent with the result of organic acid in sourdough. The single-
strain fermentation of LAB had extremely limited improvement in ester
substances, and the content and the type in LAB samples were very low.
After mixed fermentation, the types of esters increased significantly,
which indicated that mixed fermentation was beneficial to increase the
accumulation and abundance of esters. Among them, ethyl caproate,
ethyl caprylate (brandy flavor) and ethyl acetate were the three esters
with the highest content in mixed fermented sourdough. Also, ethyl
acetate had the highest content, which was a typical compound with
obvious fruity and wine aromas synthesized by ethanol and acetic acid
under the action of alcohol acyltransferase. Some studies suggested that
higher content of ethanol and acetic acid was always accompanied by
higher content of ethyl acetate (Kaseleht et al., 2011; Liu et al., 2020).
But in this study, it was found that these compounds do not have a
complete linear relationship (Pico, Bernal, & Gomez, 2015). Among the
samples, ethanol and acetic acid in LB were the lowest, while ethyl ac­
etate was the highest. The highest contents of ethyl acetate and ethyl

7
L. Fang et al. LWT 174 (2023) 114438

Table 4
Statistics on the content of volatile flavor compounds in various categories.
Sample Flavor compound content (mg/kg sourdough)

alcohols acids esters ketones aldehydes others total

Y1 1204.11 19.54 46.01 2.23 14.22 nd 1286.12

Y2 664.17 21.87 68.00 1.74 11.48 nd 767.26
L1 24.85 14.89 0.28 0.15 24.06 1.72 65.34
L2 16.35 11.01 0.17 0.10 19.51 1.11 47.70
L3 24.18 10.81 0.50 0.25 39.20 1.81 76.03
MY1L1 1057.15 52.78 79.94 0.40 42.77 nd 1234.59
MY1L2 1129.67 60.11 101.93 0.80 61.76 1.39 1355.17
MY1L3 1050.70 46.04 86.88 0.56 54.31 1.01 1241.50
MY2L1 577.65 45.01 66.50 0.37 33.61 0.27 724.15
MY2L2 502.24 42.61 65.79 0.26 34.15 nd 645.62
MY2L3 579.00 48.14 76.78 0.33 35.66 0.54 740.70
LB 221.10 16.94 118.91 nd 19.13 nd 376.34
CT0 6.58 4.92 0.12 0.52 14.63 nd 26.77

The data in the table was the quantity calculated by the internal standard. Nd, not detected.

lactate were detected in LB, reaching 79.31 mg/kg and 25.64 mg/kg,
respectively, which were significantly different from the commercial
sourdough, indicating that ethyl acetate and ethyl lactate may be the
characteristic flavor compounds of commercial sourdough LB.
4. Conclusion
In summary, the mixed fermentation of LAB and yeasts inhibited the
growth and acidification of microorganisms to a certain extent. Also, it
increased the production of acetic acid, decreased the production of
lactic acid, and improved the ratio of lactic acid content to acetic acid
content. In the sourdough system, the reducing sugars co-produced by
LAB and yeasts were used by the yeasts, which may have a positive effect
on the production of flavor compounds. Mixed fermentation had a
stronger ability to degrade phytic acid. Compared with the dough fer­
mented by LAB single strain, its total polyphenol content was reduced,
but the DPPH free radical scavenging ability was enhanced. Besides, the
types of volatile flavor compounds and the content of some compounds
increased significantly during mixed fermentation. Compared with yeast
single-strain fermented dough, the content of alcohols decreased, but the
content of acids, esters and aldehydes increased significantly. However,
the effects of homofermentative LAB on sourdough were different. Its
acidification was slower and the acetic acid content was lower. This was
accompanied by a decrease in total polyphenol content and antioxidant
properties, as well as a higher content of aldehydes. Therefore, mixed-
fermented sourdough has very good application potential.
CRediT authorship contribution statement
Luping Fang: Writing – original draft, Originally drafted, conducted
all experimental work and wrote the draft of the manuscript. Yanhua Li:
Supervision, Supervised, Writing – review & editing, reviewed and
edited the manuscript, Project administration, initiated the project and
was the main supervisor of the whole project. Weijun Wang: Supervi­
sion, Supervised, provided experimental financial support and designed
the experiment, corrected and revised the whole manuscript. Zhixia
Dou: Assisted part experimental work and revised the whole manuscript.
Jie Chen: Assisted part experimental work. Yuecheng Meng: Assisted
part experimental work.
Declaration of competing interest
The authors declare that they have no conflicts of interest.
Data availability
Data will be made available on request.
Acknowledgements
The authors are grateful to the project of High-level Innovation
Teams of Wenzhou City (20181227), Key R&D project of Zhejiang Sci­
ence and Technology Department (2019C02091).
References
Arendt, E. K., Moroni, A., & Zannini, E. (2011). Medical nutrition therapy: Use of
sourdough lactic acid bacteria as a cell factory for delivering functional biomolecules
and food ingredients in gluten free bread. Microbial Cell Factories, 10(Suppl 1), S15.
Arendt, E. K., Ryan, L., & Bello, F. D. (2007). Impact of sourdough on the texture of
bread. Food Microbiology, 24(2), 165–174.
Bartkiene, E., Vizbickiene, D., Bartkevics, V., Pugajeva, I., Krungleviciute, V.,
Zadeike, D., et al. (2017). Application of Pediococcus acidilactici LUHS29
immobilized in apple pomace matrix for high value wheat-barley sourdough bread.
LWT - Food Science and Technology, 83(15), 157–164.
Buddrick, O., Jones, O. A. H., Cornell, H. J., & Small, D. M. (2014). The influence of
fermentation processes and cereal grains in wholegrain bread on reducing phytate
content. Journal of Cereal Science, 59(1), 3–8.
Călinoiu, L. F., & Vodnar, D. C. (2018). Whole grains and phenolic acids: A review on
bioactivity, functionality, health benefits and bioavailability. Nutrients, 10(11),
1615.
Carnevali, P., Ciati, R., Leporati, A., & Paese, M. (2007). Liquid sourdough fermentation:
Industrial application perspectives. Food Microbiology, 24(2), 150–154.
Curiel, J. A., Coda, R., Centomani, I., Summo, C., & Rizzello, C. G. (2015). Exploitation of
the nutritional and functional characteristics of traditional Italian legumes: The
potential of sourdough fermentation. International Journal of Food Microbiology, 196
(3), 51–61.
Debonne, E., Schoors, F. V., Maene, P., Van Bockstaele, F., Vermeir, P., Verwaeren, J.,
et al. (2020). Comparison of the antifungal effect of undissociated lactic and acetic
acid in sourdough bread and in chemically acidified wheat bread. International
Journal of Food Microbiology, 321(16), Article 108551.
Demirkesenbicak, H., Arici, M., Yaman, M., Karasu, S., & Sagdic, O. (2021). Effect of
different fermentation condition on estimated gycemic index, in vitro starch
digestibility, and textural and sensory properties of sourdough bread. Foods, 10(3),
514–526.
Fardet, A., Rock, E., & Rémésy, C. (2008). Is the in vitro antioxidant potential of whole-
grain cereals and cereal products well reflected in vivo? Journal of Cereal Science, 48
(2), 258–276.
Fekri, A., Torbati, M., Khosrowshahi, A. Y., Shamloo, H. B., & Azadmard-Damirchi, S.
(2020). Functional effects of phytate-degrading, probiotic lactic acid bacteria and
yeast strains isolated from Iranian traditional sourdough on the technological and
nutritional properties of whole wheat bread. Food Chemistry, 306(15), Article
125620.
Filannino, P., Cagno, R. D., & Gobbetti, M. (2018). Metabolic and functional paths of
lactic acid bacteria in plant foods: Get out of the labyrinth. Current Opinion in
Biotechnology, 49(2), 64–72.
Gobbetti, M., De Angelis, M., Corsetti, A., & Di Cagno, R. (2005). Biochemistry and
physiology of sourdough lactic acid bacteria. Trends in Food Science & Technology, 16
(1–3), 57–69.
Gobbetti, M., De Angelis, M., Di Cagno, R., Calasso, M., Archetti, G., & Rizzello, C. G.
(2018). Novel insights on the functional/nutritional features of the sourdough
fermentation. International Journal of Food Microbiology, 302(8), 103–113.
Gobbetti, M., Rizzello, C. G., Cagno, R. D., & Angelis, M. D. (2014). How the sourdough
may affect the functional features of leavened baked goods. Food Microbiology, 37(2),
30–40.
Kaseleht, K., Paalme, T., Mihhalevski, A., & Sarand, I. (2011). Analysis of volatile
compounds produced by different species of lactobacilli in rye sourdough using

8
L. Fang et al.
multiple headspace extraction. International Journal of Food Science and Technology,
46(9), 1940–1946.
Katina, K., Liukkonen, K. H., Kaukovirta-Norja, A., Adlercreutz, H., Heinonen, S. M.,
Lampi, A. M., et al. (2007). Fermentation-induced changes in the nutritional value of
native or germinated rye. Journal of Cereal Science, 46(3), 348–355.
Lancetti, R., Sciarini, L., Pérez, G. T., & Salvucci, E. (2021). Technological performance
and selection of lactic acid bacteria isolated from argentinian grains as starters for
wheat sourdough. Current Microbiology, 78(1), 255–264.
Lefebvre, D., Gabriel, V., Vayssier, Y., & Fontagné-Faucher, C. (2002). Simultaneous
HPLC determination of sugars, organic acids and ethanol in sourdough process. LWT
- Food Science and Technology, 35(5), 407–414.
Liu, T. J., Li, Y., Chen, J. C., Sadiq, F. A., Zhang, G. H., Li, Y., et al. (2016). Prevalence
and diversity of lactic acid bacteria in Chinese traditional sourdough revealed by
culture dependent and pyrosequencing approaches. LWT - Food Science and
Technology, 68(5), 91–97.
Liu, T. J., Yang, L., Yang, X. Y., Yi, H. X., Zhang, L. W., & He, G. Q. (2020). The influence
of different lactic acid bacteria on sourdough flavor and a deep insight into
sourdough fermentation through RNA sequencing. Food Chemistry, 307(1), Article
125529.
Montemurro, M., Celano, G., Angelis, M. D., Gobbetti, M., Rizzello, C. G., & Pontonio, E.
(2020). Selection of non-Lactobacillus strains to be used as starters for sourdough
fermentation. Food Microbiology, 90(9), Article 103491.
Nionelli, L., Curri, N., Curiel, J. A., Cagno, R. D., Pontonio, E., Cavoski, I., et al. (2014).
Exploitation of Albanian wheat cultivars: Characterization of the flours and lactic
acid bacteria microbiota, and selection of starters for sourdough fermentation. Food
Microbiology, 44(12), 96–107.
Nistal, E., Caminero, A., Vivas, S., de Morales, J. M. R., de Miera, L. E. S., Rodríguez-
Aparicio, L. B., et al. (2012). Differences in faecal bacteria populations and faecal
bacteria metabolism in healthy adults and celiac disease patients. Biochimie, 94(8),
1724–1729.
Novotni, D., Ćurić, D., Bituh, M., Barić, C. I., Škevin, D., & Čukelj, N. (2011). Glycemic
index and phenolics of partially-baked frozen bread with sourdough. International
Journal of Food Sciences & Nutrition, 62(1), 26–33.
Pei, F., Sun, L., Fang, Y., Yang, W. J., Ma, G. X., Ma, N., et al. (2020). Behavioral changes
in glutenin macropolymer fermented by lactobacillus plantarum LB-1 to promote the
rheological and gas production properties of dough. Journal of Agricultural and Food
Chemistry, 68(11), 3585–3593.
Pico, J., Bernal, J., & Gomez, M. (2015). Wheat bread aroma compounds in crumb and
crust: A review. Food Research International, 75(9), 200–215.
LWT 174 (2023) 114438
Ravyts, F., & Vuyst, L. D. (2011). Prevalence and impact of single-strain starter cultures
of lactic acid bacteria on metabolite formation in sourdough. Food Microbiology, 28
(6), 1129–1139.
Sahin, A. W., Tom, R., Emanuele, Z., Claudia, A., Aidan, C., Lynch, K. M., et al. (2018).
Leuconostoc citreum TR116: In - situ production of mannitol in sourdough and its
application to reduce sugar in burger buns. International Journal of Food Microbiology,
302(8), 80–89.
Salim-ur-Rehman, Paterson, A., & Piggott, J. R. (2006). Flavour in sourdough breads: A
review. Trends in Food Science & Technology, 17(10), 557–566.
Settanni, L., Ventimiglia, G., Alfonzo, A., Corona, O., Miceli, A., & Moschetti, G. (2013).
An integrated technological approach to the selection of lactic acid bacteria of flour
origin for sourdough production. Food Research International, 54(2), 1569–1578.
Sidari, R., Martorana, A., Zappia, C., Mincione, A., & Giuffrè, A. M. (2020). Persistence
and effect of a multistrain starter culture on antioxidant and rheological properties of
novel wheat sourdoughs and bread. Foods, 9(9), 1258–1281.
Sripo, K., Phianmongkhol, A., & Wirjantoro, T. I. (2016). Effect of inoculum levels and
final pH values on the antioxidant properties of black glutinous rice solution
fermented by lactobacillus bulgaricus. International Food Research Journal, 23(9),
2207–2213.
Sun, L., Li, X. F., Zhang, Y. Y., Yang, W. J., Ma, G. X., Ma, N., et al. (2020). A novel lactic
acid bacterium for improving the quality and shelf life of whole wheat bread. Food
Control, 109(9), Article 106914.
Uthumporn, U., Zaidul, I. S. M., & Karim, A. A. (2010). Hydrolysis of granular starch at
sub-gelatinization temperature using a mixture of amylolytic enzymes. Food and
Bioproducts Processing, 88(1), 47–54.
Vuyst, L. D., Kerrebroeck, S. V., Harth, H., Huys, G., Daniel, H. M., & Weckx, S. (2014).
Microbial ecology of sourdough fermentations: Diverse or uniform? Food
Microbiology, 37(2), 11–29.
Wolter, A., Hager, A. S., Zannini, E., & Arendt, E. K. (2014). Influence of sourdough on in
vitro starch digestibility and predicted glycemic indices of gluten-free breads. Food &
Function, 5(3), 564–572.
Xu, D., Zhang, Y., Tang, K. X., Hu, Y., Xu, X. M., & Gänzle, M. G. (2019). Effect of mixed
cultures of yeast and lactobacilli on the quality of wheat sourdough bread. Frontiers
in Microbiology, 10(9), 2113–2125.
Xu, D., Zhang, H., Xi, J. Z., Jin, Y. M., Chen, Y. S., Guo, L. N., et al. (2020). Improving
bread aroma using low-temperature sourdough fermentation. Food Bioscience, 37
(10), Article 100704.