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Glycemic index and phenolics of partially-baked frozen bread with sourdough

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DOI: 10.3109/09637486.2010.506432

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 International Journal of Food Sciences and Nutrition,
2010; Early Online: 1–8

Glycemic index and phenolics of partially-baked frozen bread with

sourdough

DUBRAVKA NOVOTNI, DUŠKA ĆURIĆ, MARTINA BITUH, IRENA COLIĆ BARIĆ,

DUBRAVKA ŠKEVIN, & NIKOLINA ČUKELJ

Faculty of Food Technology and Biotechnology, University of Zagreb, Pierottijeva 6, HR-10000 Zagreb, Croatia
Int J Food Sci Nutr Downloaded from informahealthcare.com by 193.198.210.2 on 08/17/10

Abstract
Different lactic acid bacteria starters were used to prepare sourdough to make partially-baked frozen wholemeal wheat bread.
The sourdough was prepared with a pure culture of Lactobacillus plantarum or with commercial starters containing Lactobacillus
brevis combined with Saccharomyces cerevisiae var. chevalieri (LV4), Lactobacillus fermentum (PL1), or Lactobacillus fermentum with
phytase (PL3). We determined the acetic and lactic acid concentrations in sourdough, bread chemical composition, total
phenolics content and glycemic index (GI) in vivo. Depending on the starter, the lactic to acetic acid ratio in the sourdough was
significantly different. The GI of control bread without sourdough (70) was significantly higher than that of bread containing
sourdough prepared with LV4 starter (50), PL1 starter (56) or PL3 starter (56), but not from bread with L. plantarum
sourdough (60). The addition of 10% sourdough with a lower molar ratio of lactic to acetic acid (# 4) and higher total phenolics
content is preferable for generating bread with medium and low GI.
For personal use only.

Keywords: Glycemic index, partially-baked frozen, sourdough, wholemeal wheat bread, lactic acid bacteria

Introduction
The glycemic index (GI) is a widely used tool for
classifying carbohydrate foods according to their
ability to raise the glucose concentration in the blood
after consumption (Jenkins et al. 1981). Used in
conjunction with macronutrient and micronutrient
content, the GI guides food planning as well as
prevention and control of chronic non-infectious
diseases in many countries (FAO/WHO 1998,
Brand-Miller and Foster-Powell 1999). Cereals are a
rich source of carbohydrates that provide considerable
amounts of many nutrients in the daily diet. In fact, the
consumption of bread and pasta has the biggest effect
on the overall dietary GI of European outpatients with
type 1 diabetes (Buyken et al. 2001). The cereal
product most frequently consumed is white wheat
bread (Kulp and Ponte 2000), which causes a very
rapid rise in blood glucose level. Recently, wholegrain
breads have been attracting the interest of European
consumers as being more nutritionally balanced,
healthier, more natural and filling; in addition, they
are recognized as releasing energy more slowly and in a
slightly more digestible form than refined bread
products (Arvola et al. 2007). Wholemeal wheat
flour is richer than white wheat flour in fibers,
phenolics and minerals, but also in phytates
(Rosell et al. 2009).
At the same time, the use of innovations such as
frozen, partial-baking bread technology continues to
increase (Hamdami et al. 2007, Curic et al. 2008).
Such a two-stage baking process allows consumers to
have fresh bread any time, it decreases economical
losses due to bread staling, and it allows the
diversification of bread production. So far, partially-
baked technology has been focused mainly on white
wheat bread production. Compared with directly
baked bread, partially-baked frozen white wheat bread
has lower starch digestibility (Ronda et al. 2010) and
glycemic response (Borczak et al. 2008a). It has been

Correspondence: Duška Ćurić, PhD, Faculty of Food Technology and Biotechnology, University of Zagreb, Pierottijeva 6, HR-10000 Zagreb,
Croatia. Fax: 385 14605072. E-mail: dcuric@pbf.hr

ISSN 0963-7486 print/ISSN 1465-3478 online q 2010 Informa UK, Ltd.

DOI: 10.3109/09637486.2010.506432
 2 D. Novotni et al.
suggested that the formation of resistant starch during
freezing decreases starch digestibility (Carreira et al.
2004). Another possible explanation is that part-
baking causes a lower degree of starch granule swelling
than does conventional baking (Borczak et al. 2008b).
Moreover, starch digestibility in partially-baked bread
decreases with frozen storage time, since the increased
ice crystal formation enhances starch molecule
recrystallization (Ronda et al. 2010).
The addition of sourdough to wholemeal wheat
bread has been shown to lower the postprandial
glucose and insulin responses compared with non-
sourdough wholemeal wheat bread (De Angelis et al.
2007, Najjar et al. 2009, Scazzina et al. 2009). This
lowering is due mainly to organic acids generated by
bacteria during sourdough fermentation (Liljeberg
et al. 1995). Breads fermented with sourdough
Int J Food Sci Nutr Downloaded from informahealthcare.com by 193.198.210.2 on 08/17/10
lactobacilli showed a lower rate of starch hydrolysis
than chemically acidified breads (De Angelis et al.
2007). Sourdough has recently been added to whole-
meal wheat bread at concentrations up to 20% in order
to increase its solubility and palatability, as well as
overall nutrition value (Katina et al. 2005). The most
frequently used microorganisms for sourdough fer-
mentation are obligately homofermentative and
facultatively or obligately heterofermentative strains
For personal use only.
of lactic acid bacteria (LAB). These LAB produce
primarily lactic and acetic acids with small amounts of
citric and malic acids (Trüper and de Clari 1997).
Several antinutrients have been shown to affect the
nutritive quality of bread, including the GI (Björck and
Nyman 2006). A negative correlation between phytic
acid content and GI has been found in vitro (Björck
and Nyman 2006) and in vivo (Lee et al. 2006).
Dietary polyphenols are well-known antioxidants,
anticarcinogens or antimutagens that affect the
glycemic and insulin indices (Solomon and Blannin
2007). The major phenolics of wheat are ferulic acid,
p-coumaric acid, syringic acid, salicylic acid, cinnamic
acid, vanillic acid, caffeic acid and chlorogenic acid
(Li et al. 2008). Sourdough fermentation increases
the level of total phenolics (Coda et al. 2010) and of
free ferulic acid (Katina et al. 2007). Low doses of
ferulic acid administered orally significantly suppress
blood glucose levels in diabetic mice (Ohnishi et al.
2004) and rats (Sri Balasubashini et al. 2003). These
results indicate a need to investigate the effect of
phytates and phenolics on GI of partially-baked frozen
sourdough breads.
Currently no breads made of wholemeal wheat flour
with low GI are available (Lappi et al. 2010), including
partially-baked frozen breads. Previous studies inves-
tigated the effect of either the partially-baked freezing
process or of sourdough fermentation on the GI of the
final bread. We have developed a wholemeal wheat
bread formulation combining both sourdough fer-
mentation and partially-baked freezing technology. We
hypothesized that the application of both of these
approaches simultaneously would lower the GI of
wholemeal wheat bread more than using either
approach on its own. To test this idea, four different
LAB starters were used to ferment sourdough for use
in bread, and the glycemic response of the resulting
bread was compared with that of bread prepared by
direct yeast fermentation. Bread characteristics were
assessed after 1 month of frozen storage and after the
second baking.
Materials, subjects and methods
Wholemeal wheat flour (Granolio Ltd., Zagreb,
Croatia) had an ash content of 1.9% on dry matter
basis, a moisture content of 13.6%, wet gluten content
of 18.8%, farinograph water absorption of 64%, and
maximum viscosity of 890 AU.
Starters and growth conditions
Four starters were chosen for sourdough fermentation:
pure culture Lactobacillus plantarum DSM 2601
(DSMZ, Ltd., Braunschweig, Germany), selected as
a homofermentative strain producing mainly lactic
acid; a commercial starter of Lactobacillus brevis and
yeast Saccharomyces cerevisiae var. chevalieri
(Kvasac, Ltd., Prigorje Brdoveko, Croatia), selected
as a heterofermentative strain producing lactic and
acetic acids; and commercial starters of hetero-
fermentative Lactobacillus fermentum with phytase
(Pallazo PL3) or without phytase (Pallazo PL1),
both from Blessing Biotech, Ltd., Stuttgart, Germany.
Viable LAB cells and yeast were enumerated in
inoculum, the commercial starter and the sourdough
according to ISO 15214 (ISO 1998) and ISO 7954
(ISO 1987) standards in de Man, Rogosa and Sharpe
(MRS) medium in anaerobic conditions at 308C.
MRS broth (200 ml) was inoculated with an overnight
culture of L. plantarum (10 ml MRS broth þ LAB)
and incubated for 48 h at 308C. Cells were harvested
by centrifugation at 2,000 £ g for 10 min, washed
twice and re-suspended in 100 ml sterile tap water.
Sourdough fermentation
Sourdough was prepared by mixing wholemeal wheat
flour (900 g), tap water (900 g) and freeze-dried
commercial starter (4.5 g) or inoculum of pure culture
(100 ml) according to Dal Bello et al. (2007). The
L. plantarum inoculum contained 5.28 x 109 colony-
forming units (CFU)/ml; the PL1 starter with
L. fermentum, 5.5 £ 108 CFU/g; the PL3 starter with
L. fermentum, 5.76 £ 109 CFU/g; and the LV4 starter,
3.16 £ 1010 CFU/g (L. brevis) and 4.54 £ 108 CFU/g
(S. chevalieri). Freeze-dried starters were hydrated in
water (50 ml) at 328C for 1 hour before homogeniz-
ation with flour. Sourdough was fermented at 308C in
90% relative humidity (RH) in a proofing cabinet until
pH 4 was reached. The sourdough pH value was
 Glycemic index and phenolics of bread with sourdough
measured by pH electrode and data logger (Omega
Engineering, Inc., Stamford, CT, USA).
Determination of organic acids in sourdough
Levels of the major products of pre-fermentation,
lactic and acetic acids, were determined in sourdough
by high-performance liquid chromatography accord-
ing to Lefebvre et al. (2002). At the end of
fermentation, sourdough samples (10 g) were frozen
with liquid nitrogen and kept at 2 208C until analysis.
The sample was homogenized with approximately
60 ml distilled water and the volume adjusted to
100 ml. The solution was centrifuged for 15 min at
4,000 £ g and the supernatant was filtered through
filter paper (Whatman No. 1). A 10-ml aliquot of
filtrate was diluted with 60 ml distilled water, then
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stirred with 5 ml Carrez I solution (potassium II
hexaferrocyanate, 0.085 mol/l) and 5 ml Carrez II
solution (zinc sulfate, 0.25 mol/l). After neutralization
to pH 8.0 with 0.1 mol/l NaOH, the volume was
adjusted to 100 ml with water. The solution was
filtered through a 0.45 mm filter (Millipore filter
supplied by Labena, Ltd., Zagreb, Croatia).
The quantification was performed by a high-perform-
ance liquid chromatography system (Pro-Star 230;
Varian, Inc., Palo Alto, CA, USA) equipped with
For personal use only.
photodiode detector and an ion-exclusion column
(MetaCarb 67H, 300 £6.5 mm; Varian). The mobile
phase was 0.005 mol/l H 2SO 4; the ?ow rate,
0.6 ml/min; and the column temperature, 458C.
Test breads
Five breads were produced: four with sourdough
fermented with different starters, as well as control
bread without sourdough. Control bread without
sourdough was prepared according to Rosell et al.
(2009) by mixing wholemeal wheat flour (1,600 g),
tap water (1,024 g), compressed yeast (48 g), salt
(24 g), gluten flour (16 g) and ‘FreshBake’ improver
(16 g) (Puracor NV, Groot-Bijgaarden, Belgium).
In sourbreads, sourdough was added at the level of
10 g/100 g bread dough and the water content was
adjusted. The recipe for L. plantarum, LV4, PL1 and
PL3 sourdough breads was as follows: sourdough
(273 g), wholemeal wheat flour (1,395 g), water
(819 g), yeast (48 g), salt (24 g), gluten flour (16 g)
and improver (16 g). All ingredients were mixed in a
spiral mixer (Diosna, Ltd., Osnabrück, Germany) for
3 min at low speed and 6 min at high speed. After
resting for 10 min, bread dough was divided into 70-g
pieces, rounded, and placed in the proofing cabinet at
308C in 95% RH for 85 min. Breads were partially
baked at 1908C for 9 min with 0.2 l steam at the start,
cooled at room temperature for 60 min, frozen in an air
blast freezer set at 2 228C until the bread core reached
2 188C, and kept at 2 188C for 30 days. Partially-
baked breads were thawed at room temperature for
10 min and baked at 2108C for 6 min with 0.1 l steam.
3
Chemical analyses of the breads
The bread moisture content was determined according
to AACC 44-15A, ash content according to AACC 08-
01, proteins according to AACC 46-12 (multiplying
the nitrogen content by factor 5.7) (AACC 2000), total
dietary fiber according to AOAC method 985.29
(AOAC 2002) and total fats according to ICC Standard
136 (ICC 1984). The protein, fat, ash and moisture
content were subtracted from the total weight and the
difference was considered as total carbohydrates.
Available carbohydrates (g/100 g) were calculated as a
difference between the total carbohydrates and dietary
fiber content in bread, in accordance with the Joint
FAO/WHO Report (FAO/WHO 1998).
To determine the total phenolics content, bread
samples were cut into cubes of approximately 1 cm3
and dried at 408C for 20 h. The ethanolic extracts were
prepared as described by Yu et al. (2002a). Approxi-
mately 8 g dried sample was extracted for 3 h with
absolute ethanol using a Soxhlet extractor. The extract
was diluted with absolute ethanol in a 100-ml
volumetric flask. The total phenolics content of
ethanolic extracts was determined using the Folin –
Ciocalteau reagent (Yu et al. 2002b). The reaction was
carried out on 100 ml ethanolic extract with 500 ml
Folin – Ciocalteau reagent and 1.5 ml of 20 g/100 g
sodium carbonate. After 2 h of reaction at room
temperature, the blue color absorbance was measured
at 765 nm (HElIOS b UV-VIS Spectrometer; Pye
Unicam, Ltd., Cambridge, UK) against a blank. Gallic
acid was used as a standard. The results are expressed
as mean of triplicate measurements in mg/kg dry bread.
Participants
Ten healthy volunteers (three men and seven women),
aged 19 – 22, with an average body mass index of
22.40 ^ 0.84 kg/m2, were included in GI testing
(Table I). Volunteers were students recruited through-
out through posters placed on notice boards at the
Faculty of Food Technology and Biotechnology,
University of Zagreb. The volunteers were non-
smokers and had no history of metabolic disease, as
determined by a lifestyle questionnaire prior to
recruitment. The subjects were given full details of
the study protocol and had the opportunity to ask
questions. All subjects gave written consent prior to
participation. The volunteers were asked to maintain a
standard diet, and to abstain from heavy physical
Table I. Anthropometric characteristic of volunteers.
Weight Body mass
Gender Age (years) Height (m) (kg) index (kg/m2)
Male (n ¼ 3) 21.3 ^ 0.6 1.84 ^ 0.05 86.3 ^ 4.8 25.3 ^ 1.0
Female (n ¼ 7) 20.0 ^ 0.6 1.65 ^ 0.08 57.7 ^ 6.2 21.2 ^ 1.9
Data presented as mean ^ standard deviation.
 4 D. Novotni et al.
activity as well as consumption of alcoholic and
caffeinated drinks on the day prior to a test. The study
protocol was approved by the Ethical Committee of
the Institute for Medical Research and Occupational
Health in Zagreb, Croatia.
Study protocol
The protocol procedure was in agreement with the
recommendations of the FAO/WHO (1998), as
described by Wolever et al. (1991) and Brouns et al.
(2005). Ten subjects were included in the study, a
number that according to Brouns et al. (2005)
provides a reasonable degree of power and precision
for most purposes of measuring GI. Each subject
tested five different breads once, while the reference
food was tested twice; tests for sourdough breads and
Int J Food Sci Nutr Downloaded from informahealthcare.com by 193.198.210.2 on 08/17/10
control bread were conducted on separate days, with a
gap of at least 1 day between measurements.
Anhydrous glucose was used as a reference food.
A glucose solution was prepared by dissolving 50 g
pure glucose in 250 ml water. Each volunteer ate a
bread portion (approximately 117 g) containing 50 g
available carbohydrates over 10 min with 250 ml water.
Breads were consumed in random order on separate
occasions on the morning after a 12-h overnight fast.
For personal use only.
Capillary finger-prick blood samples were taken from
subjects at 0, 15, 30, 45, 60, 90 and 120 min after the
beginning of bread intake (time 0 min). The blood
glucose concentration was measured using an Ascensia
Contour automatic blood glucose meter (model no.
7160A; Bayer HealthCare, Leverkusen, Germany).
The blood glucose meter was checked for accuracy
using high, low and normal test solutions from the
manufacturer (Bayer Consumer Care AG, Basel,
Switzerland). Henry et al. (2007) showed strong
correlation and good agreement between results of the
Ascensia Contour blood glucose meter and other types
of clinical analyzers.
Calculations and statistical analysis
The incremental area under the curve (iAUC) of
blood glucose response, ignoring the area beneath the
baseline, was calculated geometrically (FAO/WHO
1998) using Mathematica 6.0.1 (Wolfram Research,
Inc., Champaign, IL, USA). The GI was calculated as
a ratio between mean iAUC for the bread and mean
iAUC for the glucose solution tested by the same
subject. The resulting values were averaged for all
subjects to obtain the GI value of each bread. The GI
results are expressed as mean ^ standard error.
The statistical analysis of glucose response between
five breads was made by analysis of variance (ANOVA)
for repeated measures with Bonferroni’s correction.
In addition, contrasts were performed comparing
sourdough breads with control bread.
All chemical analyses were performed in duplicate
except the total phenolics content, which was
determined in triplicate. Sourdough and bread
parameters were analyzed by ANOVA and the least-
significant difference test. All statistical analyses were
performed at P , 0.05 with Statistica 8 (StatSoft Inc.,
Tulsa, OK, USA).
Results
Sourdough and bread composition
In the present study, the obligatory heterofermentative
LAB strains L. fermentum and L. brevis, as well as the
facultatively heterofermentative L. plantarum, were
used for sourdough fermentation. At the temperature
used in this study, the fermentation time needed for
the sourdough to reach pH 4 was dependent on the
starter. L. plantarum took 23 h to decrease the pH
value, which was longer than the commercial starter
LV4 containing L. brevis in combination with yeast S.
chevalieri (15 h), the PL3 L. fermentum starter in
combination with phytase (14 h), and the PL1 starter
(20 h). The concentrations of the major fermentation
by-products in sourdough (i.e. lactic and acetic acids)
were determined at the end of fermentation. The
different starters led to sourdough with significantly
different concentration of lactic acid (P ¼ 0.032) and
acetic acid (P ¼ 0.023) (Table II). The lowest molar
ratio of lactic to acetic acid was determined in LV4
sourdough (3.06). This ratio was similar in PL1 and
PL3 sourdoughs (3.97 and 4.41, respectively), while in
L. plantarum sourdough the ratio was 126.
The macronutrient composition did not differ
significantly between breads (Table III). The breads
were a good source of protein (7.6 ^ 0.3 g/100 g or
14.1% of energy), low in fat (0.6 ^ 0.03 g/100 g), and
high in fiber (7.2 ^ 0.2 g/100 g or 3.2 g/100 kcal). The
average moisture content was 37.6 ^ 2.4 g/100 g; ash
content, 4.4 ^ 0.3 g/100 g; and available carbo-
hydrates, 42.6 ^ 2.2 g/100 g.
The addition of 10% sourdough gave significantly
higher total phenolics content than control bread
(P ¼ 0.027). The greatest increases were observed in
LV4 and PL1 sourdough breads (Table III). The total
phenolics content of PL3 and L. plantarum breads was
Table II. Lactic and acetic acid contents in sourdough at the end of
fermentation with different starters.
Lactic acid Acetic acid
Starter Microorganism (g/100 g) (g/100 g)
Pure culture L. plantarum 0.96A ^ 0.06 0.01A ^ 0.01
PL1 L. fermentum 0.88B ^ 0.05 0.15B ^ 0.03
PL3 L. fermentum 0.93C ^ 0.09 0.14B ^ 0.05
LV4 L. brevis þ 0.67D ^ 0.06 0.19C ^ 0.04
S. chevalieri
Data presented as mean ^ standard deviation of triplicate measure-
ments Means marked with different uppercase superscript letters in
the same column are significantly different (P , 0.05), according to
the least-significance test.
 Glycemic index and phenolics of bread with sourdough 5

Table III. Chemical composition of breads. of blood glucose concentration (Table IV). The mean
Sourdough bread
increase in blood glucose concentration up to 120 min
between all tested breads was significantly different
Control (P ¼ 0.009). Nevertheless, the final blood glucose
Parameter bread PL1 PL3 LV4 L. plantarum concentration at 120 min after consumption was
Moisture (g/100 g) 36.9 37.2 37.7 37.9 38.3
higher for L. plantarum bread than for control
Protein (g/100 g) 7.6 7.5 7.6 7.4 7.4 (Figure 1).
Fat (g/100 g) 0.7 0.60 0.6 0.6 0.6 Foods are classified as low, medium, or high
Ash (g/100 g) 4.1 4.7 4.3 4.2 4.1 glycemic using the following GI ranges, expressed
Fiber (g/100 g) 7.3 6.9 7.2 7.1 7.0 relative to a glucose reference: low, 55 or less; medium,
Available 43.4 43.1 42.6 42.8 42.6
carbohydrates
56 – 69; high, 70 or more (Brand-Miller and Foster-
(g/100 g) Powell 1999). The calculated GI values of sourdough
Test portion (g) 115.2 116.1 117.4 116.8 117.4 breads fermented with L. plantarum, PL1, PL3, and
Total phenolics 15.6A 19.0B 23.8C 24.2C 19.0B LV4 were, respectively, 10, 14, 14 and 20 points lower
(mg/kg dry weight) than that of control bread (Table IV). While control
Means marked with different uppercase superscript letters are signifi- bread belonged to the high-GI food group, LV4
cantly different (P , 0.05), according to the least-significance test. sourdough bread was categorized as low GI, and
Int J Food Sci Nutr Downloaded from informahealthcare.com by 193.198.210.2 on 08/17/10

L. plantarum, PL1 and PL3 were categorized as

also higher than in control bread but lower than in LV4
and PL1 breads (Table III).
Glycemic index of bread
The highest glucose response in volunteers was
observed 30 – 45 min after consumption of breads
(Figure 1). Mean blood glucose concentration was the
For personal use only.
medium GI. Three sourdough breads showed signifi-
cantly lower GIs than control bread: LV4 (P ¼ 0.019),
PL1 (P ¼ 0.014) and PL3 (P ¼ 0.018). Although
L. plantarum belonged to the medium-GI group, its GI
did not differ significantly from that of control bread
(P ¼ 0.155). The starters gave different GI values for
sourdough breads, although the differences were not
statistically significant (P ¼ 0.534).

greatest after consumption of control bread, but it did

not significantly differ among the breads (P ¼ 0.148).
At 60 min after consumption, however, the glucose
concentration was significantly lower for LV4, PL1,
and PL3 sourdough breads than for control bread
(P ¼ 0.011, P ¼ 0.020, P ¼ 0.031, respectively)
(Figure 1). The maximum increase of blood glucose
concentration, observed 45 min after eating LV4, PL1
and PL3 breads, was lower than for control bread
(P ¼ 0.035, P ¼ 0.040, P ¼ 0.049, respectively).
Consistent results were obtained for a mean increase
Discussion
The aim of the present study was to examine the effect
of combining sourdough technology and partially-
baked freezing on the GI of wholemeal wheat bread.
In addition, production of organic acids and phenolics
was compared between non-sourdough bread and
sourdough bread prepared using four different
starters for fermentation. Sourdough fermentation
with heterofermentative L. brevis and L. fermentum

Figure 1. Blood glucose response (mean value ^ standard error of the mean) in healthy volunteers during 120 min after consumption of
standard food (glucose solution, A), control bread (B) and sourdough bread fermented with L. plantarum (V), PL1 (O), PL3 ( 3 ), and LV4 (X)
starter.
 6 D. Novotni et al.
Table IV. Glucose response after consumption of test breads by 10 volunteers.
Control bread
L. plantarum
Parameter Mean SEM Mean SEM
Glucose increase (mmol/l) 1.3 0.3 1.2 0.3
Glycemic index 70 7.1 60 6.5
Data presented as mean ^ standard error of the mean (SEM). a Between all five breads. b Significantly different from control bread (P , 0.05).
produced both lactic and acetic acids, increased total
phenolic content, and gave a partially-baked frozen
wholemeal wheat bread with a GI significantly lower
than that of regular wholemeal wheat bread.
Our results show reduced glycemic response to
wholemeal wheat bread when sourdough is added. In
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previous studies, De Angelis et al. (2007), Lappi et al.
(2010), Najjar et al. (2009) and Scazzina et al. (2009)
also found significantly lower glycemic response in
sourdough bread than in wholemeal or white wheat
bread. Although our study and theirs support the same
overall conclusion, it is difficult to compare them in
detail because of the different test breads, subjects
(healthy or diabetic) and methodologies used.
The ability of sourdough to reduce blood glucose
For personal use only.
response has been ascribed mainly to organic acids
(Liljeberg et al. 1995). While LV4 sourdough in our
study had the lowest molar ratio of lactic to acetic acid,
L. plantarum produced mainly lactic acid. This result
is due to the fact that L. plantarum is a homofermen-
tative or facultatively heterofermentative bacteria
(Trüper and de Clari 1997) while L. brevis also
produces acetic acid, even more so when combined
with yeast (Meignen et al. 2001). The formation of
acids depends on the strain and other factors such as
fermentation temperature and time, dough yield and
ash content of flour (Meignen et al. 2001).
In the present study, the most significant decrease in
bread GI (29% lower than control) was observed with
LV4 sourdough, which also had the lowest lactic/acetic
acid molar ratio. Contrary to our hypothesis,
L. plantarum sourdough, which had the highest
lactic/acetic acid ratio, gave the smallest reduction in
bread GI, which was in fact statistically insignificant.
Consistent with our results, Maioli et al. (2008)
reported that glucose response was lower for sour-
dough breads prepared with a mixed culture of S.
cerevisiae, L. brevis and L. plantarum than for yeast-
fermented bread. Our results may support the
proposal by Liljeberg and Björck (1998) that the
presence of acetic acid reduces the postprandial
glucose response to bread because it reduces the
gastric emptying rate. However, studies by Najjar et al.
(2009) have called this mechanism into question.
Our finding that a lower lactic/acetic acid ratio leads
to a greater reduction in GI contrasts with several
studies. Liljeberg et al. (1995) observed significantly
Sourdough bread
PL1 PL3 LV4
Mean SEM Mean SEM Mean SEM P valuea
1.0B 0.3 1.0B 0.3 1.0B 0.3 0.009
56B 6.4 56B 5.5 50B 4.6 0.039
reduced glucose and insulin responses to wholemeal
barley bread with L. plantarum sourdough. This could
be due to a fact that barley contains considerable
amounts of b-glucan that decrease bread GI (Jenkins
et al. 2002). Östman et al. (2005) reported signifi-
cantly improved glucose tolerance, based on a 51%
reduction in total glycemic area, in hyperinsulinemic
rats who ate white wheat bread baked with addition of
lactic acid. In addition, De Angelis et al. (2007) found
lower starch hydrolysis in vitro in breads acidified with
lactic acid than in breads acidified with acetic acid.
The findings of both of these studies could be due to
the fact that they added L -lactic acid to the bread,
while in situ LAB like those in the present study
produce both L -lactic acid and D -lactic acid. Indeed,
Östman et al. (2002) reported a higher GI for L /D -
lactic acid gruel than for L -lactic acid gruel. This
question deserves further study, since to our knowl-
edge there are no reported studies on the difference
between the affect of D -lactic acid and L -lactic acid on
the GI of bread. Liljeberg et al. (1995) concluded that
lactic acid reduces postprandial glucose and insulin
responses in healthy subjects due to inhibition of
amylolytic enzymes. Östman et al. (2002) attributed
the reduced starch digestibility in the presence of lactic
acid to an interaction between starch and gluten.
PL3 and PL1 breads had the same GI value despite
the presence of phytase. It is possible that the
sourdough fermentation by L. fermentum together
with freezing process degraded phytates almost
completely (Lopez et al. 2001, Rosell et al. 2009).
We conclude that the similar organic acids compo-
sition of the sourdoughs was the main factor
determining the GI value of PL1 and PL3 breads.
Our study suggests that acidification during
sourdough fermentation is the major factor behind
the observed GI reduction. Consistent with these
results, Mettler et al. (2009) found dose-dependent
and additive effects of organic acids and phenolics on
glycemic response to food. Further work should
examine whether lactic acid or acetic acid is more
effective at reducing GI, and whether their efficacy
depends on the type of bread under consideration.
A high phenolic content in food products is
desirable because these compounds are thought to
have strong antioxidant and antimutagenic properties.
In our study, total phenolics content was significantly
 Glycemic index and phenolics of bread with sourdough
higher in breads with sourdough than in control bread.
The largest increase of phenolics was in LV4 bread,
which had 55% more phenolics than control
bread. Those results are in agreement with Coda
et al. (2010), who found the concentration of
phenolics to be higher in sourdough non-conventional
flour bread than in yeast-fermented bread. Similarly,
Katina et al. (2007) found that sourdough and yeast
prefermentation increased the levels of total phenolic
compounds and free phenolic acids of wholemeal rye.
In contrast, Lappi et al. (2010) did not observe a
significant change in the content of free phenolics such
as ferulic, sinapic and p-coumaric acids during
sourdough fermentation. Our results may differ from
those of Lappi et al. (2010) because of the difference in
the dough acidity, since the activity of the enzyme
involved in the formation of free ferulic acid is pH
Int J Food Sci Nutr Downloaded from informahealthcare.com by 193.198.210.2 on 08/17/10
dependent (Katina et al. 2007). In addition, Lappi
et al. (2010) measured free phenolics whereas we used
the Folin-C method, which measures all phenolics.
The increase in total phenolics observed in our
sourdough breads may be partly caused by an
increased amount of aromatic amino acids formed by
proteolysis during the fermentation (Katina et al.
2007).
A good inverse correlation was obtained between
For personal use only.
total phenolics content and bread GI (r ¼ – 0.877,
P ¼ 0.05). Coda et al. (2010) found that, in
comparison with baker’s yeast-fermented bread,
sourdough wholemeal wheat bread had a higher
concentration of phenolic compounds and free
amino acids, as well as a lower rate of in vitro starch
hydrolysis. It has been suggested that, like acetic acid,
phenolics lower GI by reducing the gastric emptying
rate (Hlebowicz et al. 2007).
In the present study, the control partially-baked
frozen wholemeal bread, although high fiber, exhibited
high GI. This is in agreement with the international
table of glycemic index (Foster-Powell et al. 2002).
However, we expected that freezing partially-baked
bread and storing it for 30 days at 2 188C would
decrease its GI. Indeed, Borczak et al. (2008a) found
that compared with directly baked unfrozen bread,
partially-baked frozen bread gave 28% lower GI
values. Moreover, Ronda et al. (2010) reported that
the starch digestibility of partially-baked frozen bread
decreases as a function of storage time. The difference
between our results and those of previous studies may
be that earlier work investigated the effect of freezing
on white bread, while ours is the first study to our
knowledge that examines wholemeal bread.
One limitation of this study is that the organic acids
were determined in sourdough and not in bread.
Moreover, using a larger number of subjects may
reduce the variability observed in glycemic response.
We investigated the GI of breads containing sour-
dough at 10 g/100 g, while the typical sourdough
application level is 20%. Further studies should
7
address optimal sourdough starter and dosage to
minimize bread GI.
Despite these limitations, our results suggest that
partially-baked frozen sourdough bread that has
reduced GI can be prepared using L. brevis and L.
fermentum starters. Thus, sourdough, a traditional and
natural additive, can be applied in new bread-making
technologies such as part-baking wholemeal bread to
make it a convenient part of a balanced diet for the
prevention and dietary control of diseases related to
insulin resistance.
Declarations of Interest: The present study has
been carried out with financial support from the
Commission of the European Communities, FP6,
Thematic Area ‘Food Quality and Safety’, FOOD-
2006-36302 EU-FRESH BAKE. It does not necess-
arily reflect its views and in no way anticipates the
Commission’s future policy in this area.
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