857

Utilization of different types of dietary fibres by

potential probiotics
Gui-Ying Mei, Christine M. Carey, Susan Tosh, and Magdalena Kostrzynska

Abstract: A better understanding of the functionality of probiotics and dietary fibres with prebiotic activity is required for
the development of improved synbiotic preparations. In this study, utilization of b(2–1) fructans, galactooligosaccharides,
Can. J. Microbiol. Downloaded from www.nrcresearchpress.com by CONCORDIA UNIV on 03/19/13

and plant polysaccharides as prebiotics by lactobacilli, bifidobacteria, and pediococci was investigated. Our results demon-
strate that prebiotics with linear chains consisting of galactose units are better utilized by probiotics than are those consisting
of glucose and fructose units, and the ability of probiotic bacteria to utilize prebiotics is strain-specific. In addition, rye
fructooligosaccharides represent a prebiotic fibre that supports the growth of a wide range of probiotic cultures and as such
has a potential to improve the successfulness of probiotic treatments. This study also demonstrates dietary fibre utilization
by pediococci and provides data supporting the possible use of pediococci as a probiotic in synbiotic combinations.
Key words: probiotics, prebiotics, oligosaccharides, polysaccharides, lactic acid bacteria.
Résumé : Une meilleure connaissance de la fonctionnalité des probiotiques et des fibres alimentaires possédant une activité
prébiotique est requise afin de développer de préparations synbiotiques améliorées. Dans cette étude, l’utilisation comme
prébiotiques de b(2–1) fructanes, de galacto-oligosaccharides et de polysaccharides végétaux par les lactobacilles, les bifido-
bactéries et les pédiocoques a été examinée. Nos résultats démontrent que les prébiotiques possédant des chaines linéaires
formées d’unités de galactose sont mieux utilisées par les probiotiques comparativement à celles qui consistent en unités de
glucose et de fructose, et que la capacité des bactéries probiotiques à utiliser les prébiotiques est spécifique à la souche. De
For personal use only.

plus, le fructooligosaccharides de seigle représente une fibre prébiotique qui permet la croissance d’une vaste gamme de
cultures probiotiques et de ce fait, a le potentiel d’améliorer le succès des traitements aux probiotiques. Cette étude démon-
tre aussi que les pédiocoques utilisent les fibres alimentaires et présente des données qui appuient l’utilisation possible du
genre Pediococcus comme probiotique dans des combinaisons synbiotiques.
Mots‐clés : probiotiques, prébiotiques, oligosaccharides, polysaccharides, bactéries lactiques.
[Traduit par la Rédaction]

Introduction cherichia coli O157:H7 with P. pentosaceus or Pediococcus

acidilactici results in downregulation of the Shiga-toxin 2 A
Probiotics and prebiotics have been extensively investi- (stx2A) gene (Carey et al. 2008). The possible probiotic
gated in recent years for their potential beneficial effects on mechanisms promoting human health include supplying of
human health and well-being. Probiotics are defined as live nutrients to the host, preventing pathogen growth, restoring
microorganisms that confer a health benefit on the host microbial homeostasis, competing and cooperating for nu-
when administrated in adequate amounts (Food and Agricul- trients, producing antimicrobial compounds such as H2O2
ture Organization of the United Nations – World Health Or- and bacteriocins, inhibiting intestinal inflammation, prevent-
ganization 2002). There are extensive data to indicate that ing certain cancers, and stimulating the immune system
certain probiotics, such as Lactobacillus and Bifidobacterium (Marco et al. 2006; Lebeer et al. 2008; Gupta and Garg
spp., are critical for human health (Collins and Gibson 1999). 2009; Ng et al. 2009). Apart from above mechanisms, there
The pediococci are a diverse group of lactic acid bacteria is evidence that Lactobacillus salivarius inhibits growth of
(LAB) some of which have biotechnological importance Helicobacter pylori not through acid production or bacterio-
(Raccach 1987). In addition, animals treated with an LAB cins or bacteriocin-like peptides but through alternative
mixture containing Pediococcus pentosaceus have a reduced mechanisms that require the presence of live cells. In addi-
incidence, severity, and duration of diarrhea, demonstrating tion, growth inhibition of H. pylori by L. salivarius is strain-
the potential use of pediococci as probiotics (Casey et al. dependent (Ryan et al. 2008). Alternatively, consumption of
2007). A recent study also showed that co-incubation of Es- food ingredients, such as prebiotics, can also be used to mod-
Received 4 May 2011. Revision received 23 June 2011. Accepted 29 June 2011. Published at www.nrcresearchpress.com/cjm on
29 September 2011.
G.-Y. Mei. Agriculture and Agri-Food Canada, Guelph Food Research Center, 93 Stone Road West, Guelph, ON N1G 5C9, Canada;
Department of Plant Pathology, China Agricultural University, Beijing 100193, People’s Republic of China.
C.M. Carey, S. Tosh, and M. Kostrzynska. Agriculture and Agri-Food Canada, Guelph Food Research Center, 93 Stone Road West,
Guelph, ON N1G 5C9, Canada.
Corresponding author: Magdalena Kostrzynska (e-mail: kostrzynskam@agr.gc.ca).

Can. J. Microbiol. 57: 857–865 (2011) doi:10.1139/W11-077 Published by NRC Research Press
 858 Can. J. Microbiol. Vol. 57, 2011

ulate the colonic microbiota and improve host health. A pre- Table 1. Probiotic strains used in this study.
biotic is a selectively fermented food ingredient that stimu-
ATCC or FRP
lates specific changes in the composition of and (or) activity
Strain No.a Source of isolation
in the gastrointestinal microflora, which in turn confer benefits
Lactobacillus spp.
on the host well-being and health (Roberfroid 2007). Prebiot-
L. rhamnosus GG ATCC 53103 Human feces
ics, such as fructooligosaccharides (FOS), inulin, galactooligo-
L. curvatus FRP 15 German sausage
saccharides (GOS), and glucooligosaccharides, are dietary
L. plantarum FRP 16 Dairy products
carbohydrates that escape digestion in the upper gastrointesti-
L. reuteri ATCC 23272 Human feces
nal tract and alter the bacterial composition of the gut by
L. kefir IM002 Dairy products
changing the type of the substrate provided to the existing gut
microbiota (Gibson and Roberfroid 1995; Collins and Gibson
Bifidobacterium spp.
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1999; Alander et al. 2001; Malinen et al. 2002).

B. adolescentis ATCC 15703 Intestine of adult
Prebiotics are fermented by probiotic bacteria, producing B. longum ATCC 15707 Intestine of adult
short-chain fatty acids (SCFAs) as the end products of fer- B. longum ATCC 15708 Intestine of adult
mentation. The major SCFAs produced are acetate, propio- B. breve FRP 334 Human infant
nate, and butyrate (Pylkas et al. 2005), and the profile of
SCFAs varies among the different prebiotic sources. SCFAs Pediococcus spp.
have beneficial effects on human health as energy sources P. acidilactici FRP 236 Meat-associated
and by inhibiting the growth of pathogenic bacteria (Blottiere P. pentosaceus FRP 243 German sausage
et al. 1999; Pylkas et al. 2005). SCFAs also decrease colonic P. pentosaceus FRP 244 German sausage
pH, which demonstrates the fermentability of the nondigesti- a
ble carbohydrates. ATCC, American Type Culture Collection; FRP designates the Guelph
Food Research Centre culture collection of Agriculture and Agri-Food Canada.
Currently, research has shown that prebiotics can signifi-
cantly modify the composition of the gut microbiota both in CO2 at 37 °C in modified de Mann – Rogosa – Sharpe broth
vitro and in vivo (Gibson and Roberfroid 1995). Kaplan and (de Mann – Rogosa – Sharpe supplemented with 0.2 g of so-
Hutkins (2000) screened 28 LAB for their ability to ferment dium carbonate and 0.1 g of calcium choride in 1 L distilled
For personal use only.

inulin and oligofructose and reported that 12 of 16 Lactoba-
cillus strains and 7 of 8 Bifidobacterium strains tested were
able to ferment the substrates. Several studies have demon-
strated that inulin and oligofructose are selectively fermented
by bifidobacteria (Roberfroid et al. 1998; Perrin et al. 2002;
Oliveira et al. 2009, 2011b). Jaskari et al. (1998) reported
that b-glucooligomers (from oat) and raffinose enhance the
growth of health-promoting probiotic strains but only raffi-
nose does it at a significant level. In addition, a comparative
study showed that GOS were effective at increasing the num-
ber of bifidobacteria and lactate while generating less gas
(Rycroft et al. 2001). It has been also shown that oral admin-
istration of GOS enhances faecal levels of Bifidobacterium
animalis subsp. lactis (Malinen et al. 2002). Kedia et al.
(2008) evaluated the fermentability of oat fractions obtained
by debranning using three lactobacilli and demonstrated that
a 1%–3% pearling fraction, containing the highest concentra-
tion of total dietary fibre compared with other pearling frac-
tions, is the most suitable for fermentation and produces
considerably higher probiotics cell concentrations.
The objective of this study was to examine the utilization
of a number of plant oligosaccharides and polysaccharides as
prebiotics (which are characterized by different linear chain
components) by selected probiotic bacteria. This will provide
important information about the factors that influence the fer-
mentation of nondigestible carbohydrates by the probiotic
bacteria and information that can be used to determine which
types of prebiotics are suitable to prepare new and improved
prebiotic–probiotic formulations.
Materials and methods
Bacterial strains and growth conditions
The bacterial strains used in this study are listed in Table 1.
Lactobacillus spp. and Pediococcus spp. were grown in a 5%
water and 5% filter-sterilized L-cysteine HCl monohydrate
(FisherBiotech)) for 24 h. L-Cysteine was added to provide
amino nitrogen as a growth factor and to reduce redox poten-
tial. Bifidobacterium strains were grown in Reinforced Clos-
tridial Medium broth (Oxoid) at 37 °C for 48 h under
anaerobic atmosphere using GasPak EZ Anaerobe Container
System Sachets (Becton Dickinson and Company, Maryland,
USA) in the GasPak EZ Incubation Container (Becton Dick-
inson and Company). Dry anaerobic indicators strips (Becton
Dickinson and Company) were used to verify that an anaero-
bic atmosphere was attained.
Prebiotics
Sixteen different types of dietary fibers were examined.
The carbohydrates, degree of polymerization, and where ap-
propriate, the commercial source are shown in Table 2. Glu-
cose and galactose, used as controls, were purchased from
Sigma.
Basal medium used for prebiotics studies
The basal medium (Degnan and Macfarlane 1995) was
used as a carbohydrate-free medium for testing the effect of
the potential prebiotics on the growth of the probiotic strains.
The basal medium consisted of (g/L) peptone water, 5.0;
tryptone, 10; yeast extract, 2.5; Tween 80, 1.0; NaCl, 4.5;
KCl, 0.25; MgCl2·6H2O, 0.15; KH2PO4, 0.40; K2HPO4,
0.20; NH4Cl, 0.40; and cysteine–HCl, 0.50.
Preparation of cell suspensions
In preparation for the fermentation trials, the bacteria were
grown in 10 mL of the appropriate medium (modified
de Mann – Rogosa – Sharpe or Reinforced Clostridial Me-
dium) containing glucose (10 g/L) as the carbon source.
After incubation, the bacterial cells were collected by centri-

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 Mei et al. 859

Table 2. Prebiotics and galactose (control) tested in this study.

Substrate Chemical structure Source Manufacturer

Fructooligosaccharides Mixture of 1-kestosea; 1,1-kestotetraoseb; Rye GFRC
1,1,1-kestopentaosec; and sucrosed
Fructooligosaccharides (2→1)-b-Glucosyl-fructose)n, where n is 2–60 Chicory Sigma
Inulin Heterogeneous blend of fructose polymers Chicory Sigma
Galactose D-Galactose Sigma
b-Glucotriose 3-o-b-Glucosyl-D-cellobiose Barley Megazyme
b-Glucotetrose 3-o-b-Glucosyl-D-cellotriose Barley Megazyme
b-Glucopentose 3-o-b-Cellotetraosyl-D-glucose Barley Megazyme
b-Glucohexose 3-o-b-Cellopentaosyl-D-glucose Barley Megazyme
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Arabinan (1→3)(1→5)-a-Arabinose Sugar beet Megazyme

b-Glucan (high viscosity) (1→3)(1→4)-b-D-Glucan Oat bran GFRC
b-Glucan (medium viscosity) (1→3)(1→4)-b-D-Glucan Oat Megazyme
b-Glucan (low viscosity) (1→3)(1→4)-b-D-Glucan Barley Megazyme
Arabinoxylan (low viscosity) (1→4)-b-D-Xylopyranose substituted with Wheat Megazyme
arabinose at carbon 2, 3, or both
Arabinoxylan (high viscosity) (1→4)-b-D-Xylopyranose substituted with Rye Megazyme
arabinose at carbon 2, 3, or both
Galactan b-(1→4)-Galactose Potato Megazyme
Galactooligosaccharides Mixture of stachyosee, raffinosef, and sucrose Soy GFRC
Oligomate 55NP b-(1→4)-Galactosyllactose Milk Yakult
Oligomate 55N b-(1→4)-Galactosyllactose Milk Yakult
Note: GFRC, Guelph Food Research Centre.
a
1-Kestose — O-b-D-fructofuranosyl-(2–1)-b-D-fructofuranosyl-(2–1)-a-D-glucopyranoside.
For personal use only.

b
1,1-Kestotetraose — O-b-D-fructofuranosyl-(2–1)-b-D-fructofuranosyl-(2–1)-b-D-fructofuranosyl-a-D-glucopyranoside.
c
1,1,1-Kestopentaose — O-b-D-fructofuranosyl-(2–1)-b-D-fructofuranosyl-(2–1)-b-D-fructofuranosyl-(2–1)-b-D-fructofuranosyl-a-D-glucopyranose.
d
Sucrose — b-D-fructofuranosyl-a-D-glucopyranoside.
e
Stachyose — b-D-fructofuranosyl-O-a-D-galactopyranosyl-(1→6)-O-a-D-galactopyranosyl-(1→6)-a-D-glucopyranoside.
f
Raffinose — O-a-D-galactopyranosyl-(1→6)-a-D-glucopyranosyl-b-D-fructofuranoside.

fugation at 5000g for 10 min at 10 °C, washed once with
physiological saline (0.85% NaCl solution), and resuspended
in 10 mL of basal medium to remove excess carbon prior to
the fermentation trials.
In vitro fermentation
Growth of the probiotic strains was monitored in basal me-
dium supplemented with 1% of each test prebiotic as the sole
carbon source (Table 2). Following media preparation, a 1%
prewashed inoculum of a given probiotic culture was used to
examine the effect of the selected prebiotics on the growth of
the probiotic culture. Cultures (10 mL) were incubated at
37 °C for a period of 48 h under anaerobic conditions. Each
fermentation experiment was performed in triplicate. The
controls consisted of inoculated basal medium lacking prebiotics
and inoculated basal medium containing glucose or galactose.
The growth of each strain was monitored at stationary cul-
ture phase at 48 h by measuring the optical density (OD) of
the cultures at 600 nm using a spectrophotometer (Ultrospec
3100 pro).
The pH of aliquots was also determined at stationary cul-
ture phase (48 h) using a pH meter (Fisher Scientific), soon
after removing the samples from the fermentation vessels.
Data and statistical analysis
For each experiment, the data were analyzed using the Ex-
cel statistical package. The OD and pH readings and corre-
sponding standard deviations were calculated from triplicate
samples from three separate experiments. Statistical signifi-
cance between treatment and control conditions was assessed
by the Tukey’s test. A significant difference was defined as a
P value of <0.05.
Results
FOS from rye and chicory greatly promoted the growth of
all the strains tested (Fig. 1). Consequently, the bacterial me-
tabolism of these substrates caused a marked decrease in the
culture medium pH (Fig. 1). All the Bifidobacterium strains
tested and P. pentosaceus FRP 243 also showed a great abil-
ity to utilize inulin from chicory root (Figs. 1B and 1C).
No influence of b-glucan (from either oat or barley) or
other polysaccharides on the growth of probiotics or the pH
of culture medium was observed (Fig. 2); whereas, all the pro-
biotics tested grew very well in b-glucotriose (3-o-b-glucosyl-
D-cellobiose prepared by enzymatic hydrolysis of barley b-
glucan using lichenase (Megazyme)), and the corresponding
pH of the medium from fermentation significantly decreased
(P < 0.05) (Fig. 3). With the exception of Lactobacillus
curvatus (FRP 15) and P. acidilactici (FRP 236), all other
strains were able to utilize and ferment b-glucotetrose (3-o-
b-glucosyl-D-cellotriose) prepared by enzymatic hydrolysis
of barley b-glucan (Megazyme). Lactobacillus rhamnosus
GG (ATCC 53103), L. kefir IM002, B. breve (FRP 334),
and P. pentosaceus (FRP 244) grew well in b-glucopentose
(3-o-b-cellotetraosyl-D-glucose) (Megazyme). Bifidobacterium
breve (FRP 334) was the only probiotic strain that demon-
strated an ability to utilize b-glucohexose (3-o-b-cellopen-

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 860 Can. J. Microbiol. Vol. 57, 2011

Fig. 1. The utilization of b(2–1) fructan-type prebiotics by lactoba-

cilli (A), bifidobacteria (B), and pediococci (C). Probiotic bacteria
were incubated in the basal medium containing 1% chicory inulin
and 1% fructooligosaccharides (FOS) from chicory and rye at 37 °C
for 48 h, and the OD600 and pH of cultures were measured. Basal
medium and medium containing 1% glucose were used as controls.
The graphs represent the results of three experiments; * denotes a
significant difference between treatments and control basal medium
samples (P < 0.05).

taosyl-D-glucose) prepared by enzymatic hydrolysis of bar-

ley b-glucan (Megazyme) (Fig. 3). The pH of the culture
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medium in which those probiotic bacteria grew well de-

creased correspondingly (Fig. 3).
GOS (from soy), Oligomate 55NP (powder), and Oligo-
mate 55N (syrup) significantly (P < 0.05) enhanced the
growth of all the probiotics tested, while galactan had very
little effect on the growth of probiotics (Figs. 4A–4C, top
panels). Consequently, the bacterial metabolism of oligomates
caused a marked decrease in the culture medium pH
(Figs. 4A–4C, bottom panels). Though GOS from soy signif-
icantly improved the growth of all tested probiotics, the de-
crease of pH was not so significant compared with cultures
grown in media with oligomates or galactose.
The results of this study also demonstrate that the ability
of probiotic bacteria to utilize prebiotics varies even within
For personal use only.

the same species.

Discussion
To determine which prebiotics were utilized by probiotics,
including bifidobacteria, lactobacilli, and pediococci, three
types of dietary fibres, based on the different composition of
linear chains, were evaluated for their effect on the growth of
12 probiotic strains and on fermentation. b(2–1) Fructans are
a category of nutritional compounds in which one or more
fructosyl-fructose linkages comprise the majority of glycosi-
dic bonds, with or without a terminal a(1–2)-linked D-glucose,
including FOS and inulin (Roberfroid et al. 1998; Kaplan
and Hutkins 2000; Kelly 2008). In our study, the b(2–1)
fructans that were evaluated include FOS from rye and inu-
lin and FOS from chicory. This is the first time that the uti-
lization of FOS from rye by probiotic bacteria has been
reported, and the results demonstrate that FOS from rye is
utilized by probiotics. In addition, bacteria grew better on
FOS from rye than on FOS from chicory. Only four tested
Bifidobacterium strains and P. pentosaceus (FRP 243), but
not P. pentosaceus (FRP 244), fermented inulin from chicory.
The difference between the two P. pentosaceus strains in their
utilization of inulin may suggest strain-specificity (Fig. 1C).
According to the results, the b(2–1) fructans with short
chains greatly support growth of the probiotics, as com-
pared with the longer-chained b(2–1) fructans, and all b(2–1)
fructans are bifidogenic (Gibson and Wang, 1994a; Modler
1994; Roberfroid et al. 1998; Kelly 2008: Oliveira et al.
2009, 2011b). In our study, bifidobacteria appear to grow
better on FOS than on glucose (Fig. 1B), which is consis-
tent with results reported by Gibson and Wang (1994b)
and Roberfroid et al. (1998), indicating that bifidobacteria
preferentially metabolize short FOS fractions due to the
presence of appropriate uptake systems (Van der Meulen et
al. 2004, 2006).

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 Mei et al. 861

Fig. 2. Utilization of various polysaccharides by Lactobacillus spp.

(A), Bifidobacterium spp. (B), and Pediococcus spp. (C). Probiotic
bacteria were incubated in basal medium containing polysaccharides
at 37 °C for 48 h, and the OD600 and pH of cultures were measured.
The graphs represent the results of three replicates; * denotes a sig-
nificant difference between treatments and control basal medium
samples (P < 0.05).

The second group of prebiotics tested includes b-glucans

from barley and oats, and b-glucooligomers (glucooligosac-
charides prepared from barley b-glucan), which consist of al-
ternate b-(1,3)-/b-(1,4)-linked glucosyl residues. None of the
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strains grew in basal medium containing b-glucans, particu-

larly high-viscosity b-glucans from oat (Fig. 2). All probiotics
assayed were able to utilize b-glucotriose, while the growth
rate on b-glucotetrose decreased. Only four strains, L. rhamno-
sus GG (ATCC 53103), L. kefir IM002, B. breve (FRP 334),
and P. pentosaceus (FRP 244), could use b-glucopentose, and
only B. breve (FRP 334) utilized b-glucohexose (Fig. 3).
The results obtained provide evidence that the degree of
polymerization (DP) influences the ability of probiotic bac-
teria to utilize the b-glucooligomers. In addition to our
study, Snart et al. (2006) reported that some lactobacilli uti-
lize oligosaccharides (DP3 or DP4) present in barley b-glucan
hydrolysates, and clearly demonstrated that high-viscosity
b-glucan can produce a prebiotic effect in the ceca of rats.
For personal use only.

According to our results, not only lactobacilli but also bifi-

dobacteria and pediococci can use purified barley b-glucan
hydrolysates, especially b-glucotriose, which was used by
all of the strains. As such, the present study further con-
firmed the benefit of using b-glucan products as dietary
supplements for human consumption.
In addition, recent studies by Grimoud et al. (2010) have
demonstrated that bifidobacteria can grow on oligoalternan,
which consists of alternate a-(1,3)-/a-(1,6)-linked glucosyl
residues (DP3 to DP6) and on oligodextran consisting of
a-(1,6)-linked glucosyl residues (DP3 to DP9). No significant
growth rates of lactobacilli on oligoalternan were observed,
and only three of the lactobacilli were able to use oligo-
dextran. The results suggest that not only the DP of glucooli-
gosaccharides but also the linkage of the glucosyl residues
influence the utilization of the prebiotics by probiotic strains.
Arabinan from sugar beet and pentosan arabinoxylans from
wheat and rye were also tested in our study. None of these
polysaccharides promoted the growth of the probiotic strains
investigated. Arabinoxylans (AXs) are the major non-starch
polysaccharides present in many cereal grains. Recent studies
revealed positive effects of AXs on cecal fermentation
(Hopkins et al. 2003), and arabinoxylan oligosaccharides
(AXOS) preparation with an average DP (avDP) of 5 and an
average degrees of arabinose substitution of 0.27 exhibit the
best combination of desirable effects on gut health character-
istics (Van Craeyveld et al. 2008). Although some health ef-
fects of AXs are reported, the effects of their hydrolysis
products, AXOS, are less studied because of their variable
DP and arabinose substitution, i.e, AXOS compounds have a
much higher diversity than other commercial oligosacchar-
ides (Grootaert et al. 2007).
The last group of prebiotics tested included Oligomate 55
(powder and syrup, commercial prebiotic GOS) and GOS
from soy. The prebiotics listed all have a linear chain that

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 862 Can. J. Microbiol. Vol. 57, 2011

Fig. 3. The growth (OD600) of Lactobacillus spp. (A), Bifidobacter-

ium spp. (B), and Pediococcus spp. (C), and pH of culture media
containing short-chain oligosaccharides prepared by enzymatic hy-
drolysis of barley b-glucan. The graphs represent the results of three
experiments; * denotes a significant difference between treatments
and control basal medium samples (P < 0.05).

comprises one or more galactose units. Increases in growth

rate in media with these substrates were observed for all the
probiotic bacteria tested. The present results are not only con-
sistent with previous findings, that bifidobacteria utilize
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GOS, which belong to “Bifidus factor” candidates (Sako et

al. 1999; Alander et al. 2001; Vernazza et al. 2006; Barboza
et al. 2009; Oliveira et al. 2011a), but also indicate that all
the lactobacilli and pediococci assayed in our study utilized
GOS. Previous reports concerning GOS focussed on the prebi-
otic properties of GOS produced from lactose in human milk
and to a smaller extent in cow’s milk using b-galactosidases
isolated from different sources (Alander et al. 2001; Rabiu
et al. 2001; Tannock et al. 2004; Tzortzis et al. 2005;
González et al. 2008; Jung et al. 2008). The commercial
products Oligomate 55N and Oligomate 55NP are made
from lactose in milk using b-galactosidase derived from
Kluyveromyces lactis (Kobayashi et al. 2009). There has
been very little research focussing on GOS from other
For personal use only.

sources, particularly plant sources. Our study illustrates that

GOS from plant source (soy) can be an efficient prebiotic
for many beneficial bacteria. Though the GOS from soy
was found to be the most efficient prebiotic in promoting
the growth of probiotics assayed, the pH was not as low as in
the case of Oligomate 55N and 55NP and galactose (Fig. 4).
The pediococci are a group of gram-positive homofermen-
tative LAB (Gonzalez and Kunka 1983), and recent studies
have demonstrated the potential use of pediococci as a probi-
otic candidate (Casey et al. 2007; Carey et al. 2008). How-
ever, studies associated with the ability of pediococci to use
prebiotics are very limited. The only evidence available is the
ability to ferment the trisaccharide raffinose by strains of
P. pentosaceus and sucrose by P. acidilactici PAC1.0
(Gonzalez and Kunka 1986, 1987). In this study, we investi-
gated the utilization of 16 prebiotics by pediococci. Three
strains of Pediococcus spp. (P. acidilactici FRP 236, P. pen-
tosaceus FRP 243, and P. pentosaceus FRP 244) were as-
sayed, and the results indicated the ability of these strains to
utilize prebiotics (Figs. 1C, 2C, 3C, and 4C). This ability was
strain-specific and may be associated with the presence of
plasmids. As such, our results demonstrate that selected Ped-
iococcus strains have potential probiotic properties and could
be used in preparing pre-probiotic formulations.
Previous papers dealing with fermentation of mono- and
oligo-saccharides by bifidobacteria report wide differences in
sugar preferences among oligosaccharides and their mono-
meric constituents (Mlobeli et al. 1998; Palframan et al.
2002, 2003; Van der Meulen et al. 2004, 2006). In the
present study, oligosaccharides were the preferred sugar over
glucose in terms of growth rate in L. kefir IM002, B. adoles-
centis ATCC 15703, B. longum ATCC 15708, B. breve FRP
334, P. pentosaceus FRP 243, and P. pentosaceus FRP 244
in all types of prebiotics tested. While glucose was the pref-
erence sugar over all types of tested oligosaccharides in the

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 Mei et al. 863

Fig. 4. The utilization of galactose, galactooligosaccharides (GOS)

from milk and soy, and galactan by Lactobacillus spp. (A), Bifido-
bacterium spp. (B), and Pediococcus spp. (C). Probiotic bacteria
were incubated in basal medium containing the saccharides at 37 °C
for 48 h, and the OD600 and pH of cultures were measured. The
graphs represent the results of three replicates; * denotes a signifi-
cant difference between treatments and control basal medium sam-
ples (P < 0.05).

growth of L. reuteri ATCC 23272. Lactobacillus rhamnosus

GG ATCC 53103 grew to the highest OD on glucose com-
pared with FOS and b-glucotriose. Lactobacillus curvatus
Can. J. Microbiol. Downloaded from www.nrcresearchpress.com by CONCORDIA UNIV on 03/19/13

FRP 15 and L. plantarum FRP 16 prefer oligosaccharides

over glucose in GOS and glucooligosaccharides. GOS were
the preferred sugar over monosaccharides in the growth of
P. acidilactici FRP 236. Bifidobacterium longum ATCC
15707 grew better on FOS and GOS than on glucose. Based
on the results of our study, carbohydrate preferences of pro-
biotic bacteria are widely different even within a single strain
given the different types of prebiotics. But the report by
Amaretti et al. (2007) that “monosaccharides were preferred
over oligosaccharides in a few cases, while growth rates and
cell yields were higher on oligosaccharides than on their
monomeric moieties in many others” is still reliable. The
preferential consumption of oligosaccharides of most probi-
otic strains (10 out of 12) can be an advantage for the strains
For personal use only.

in their competition for nutrition with other microorganisms

in the human gut, where oligosaccharides are the main sugars.
The reduction in pH of culture medium in which the
growth of probiotics is promoted illustrates that the SCFAs
may be produced. SCFAs are end products of prebiotics fer-
mentation and are beneficial to human health, as they can de-
crease colonic pH and inhibit non-acid-tolerant bacteria while
supporting the growth of beneficial microbiota in gut. They
may also contribute toward host energy requirements.
Previous studies focussing on in vitro fermentation of inu-
lin indicated that molecules with a chain length (DP) of >10
are fermented on average half as quickly as molecules with a
DP of <10 (Roberfroid et al. 1998). Sanz et al. (2005) re-
ported that disaccharides with linkages of 1–2, 1–4, and 1–6
generate a high prebiotic index. In our study, the fermentabil-
ity of two types of prebiotics b(2–1) fructans and b-
glucooligosaccharides appear to be determined by chain
length (DP); whereas, the fermentability of prebiotics with
linear chains consisting of galactose units (GOS-type) is not
affected by the chain length, which illustrates that linear
chains consisting of galactose units are prone to be used by
probiotics compared with those consisting of glucose and
fructose units. This structure–component–fermentability in-
formation obtained in this study may lead to a predictive
understanding of how specific structures of prebiotics are fer-
mented by the human gut microbiota and may provide infor-
mation in the studies of dietary modulation of probiotic
bacteria. These results may also help to design new improved
prebiotic–probiotic preparations.

Acknowledgements
Financial support by the Agriculture and Agri-Food Canada
(AAFC) and China Scholarship Council is gratefully acknowl-
edged. We thank Dr. E.R. Farnworth (AAFC) for providing
Lactobacillus kefir and Yakult, Tokyo, Japan for Oligomate 55.

Published by NRC Research Press

 864
References
Alander, M., Matto, J., Kneifel, W., Johansson, M., Kogler, B.,
Crittenden, R., et al. 2001. Effect of galacto-oligosaccharides
supplementation on human faecal microflora and on survival and
persistence of Bifidobacterium lactis Bb-12 in the gastrointestinal
tract. Int. Dairy J. 11(10): 817–825. doi:10.1016/S0958-6946(01)
00100-5.
Amaretti, A., Bernardi, T., Tamburini, E., Zanoni, S., Lomma, M.,
Matteuzzi, D., and Rossi, M. 2007. Kinetics and metabolism of
Bifidobacterium adolescentis MB 239 growing on glucose,
galactose, lactose, and galactooligosaccharides. Appl. Environ.
Can. J. Microbiol. Downloaded from www.nrcresearchpress.com by CONCORDIA UNIV on 03/19/13
Microbiol. 73(11): 3637–3644. doi:10.1128/AEM.02914-06.
PMID:17434997.
Barboza, M., Sela, D.A., Pirim, C., LoCascio, R.G., Freeman, S.L.,
German, J.B., et al. 2009. Glycoprofiling bifidobacterial con-
sumption of galacto-oligosaccharides by mass spectrometry
reveals strain-specific, preferential consumption of glycans.
Appl. Environ. Microbiol. 75(23): 7319–7325. doi:10.1128/
AEM.00842-09. PMID:19801485.
Blottiere, H.M., Champ, M., Hoebler, C., Michel, C., and Cherbut, C.
1999. Production and digestive effects of short chain fatty acids.
Sci. Aliments, 19: 269–290.
Carey, C.M., Kostrzynska, M., Ojha, S., and Thompson, S. 2008. The
effect of probiotics and organic acids on Shiga-toxin 2 gene
expression in enterohemorrhagic Escherichia coli O157:H7. J.
Microbiol. Methods, 73(2): 125–132. doi:10.1016/j.mimet.2008.
For personal use only.
01.014. PMID:18328583.
Casey, P.G., Gardiner, G.E., Casey, G., Bradshaw, B., Lawlor, P.G.,
Lynch, P.B., et al. 2007. A five-strain probiotic combination
reduces pathogen shedding and alleviates disease signs in pigs
challenged with Salmonella enterica serovar Typhimurium. Appl.
Environ. Microbiol. 73(6): 1858–1863. doi:10.1128/AEM.01840-
06. PMID:17261517.
Collins, M.D., and Gibson, G.R. 1999. Probiotics, prebiotics, and
synbiotics: approaches for modulating the microbial ecology of the
gut. Am. J. Clin. Nutr. 69(5): 1052S–1057S. PMID:10232648.
Degnan, B.A., and Macfarlane, G.T. 1995. Arabinogalactan utiliza-
tion in continuous cultures of Bifidobacterium longum: effect of
co-culture with Bacteroides thetaiotaomicron. Anaerobe, 1(2):
103–112. doi:10.1006/anae.1995.1005. PMID:16887514.
Food and Agriculture Organization of the United Nations – World
Health Organization. 2002. Guidelines for the evaluation of
probiotics in food [online]. Available from ftp://ftp.fao.org/es/esn/
food/wgreport2.pdf [accessed 4 August 2010].
Gibson, G.R., and Roberfroid, M.B. 1995. Dietary modulation of the
human colonic microbiota: introducing the concept of prebiotics.
J. Nutr. 125(6): 1401–1412. PMID:7782892.
Gibson, G., and Wang, X. 1994a. Bifidogenic properties of different
types of fructo-oligosaccharides. Food Microbiol. 11(6): 491–498.
doi:10.1006/fmic.1994.1055.
Gibson, G.R., and Wang, X. 1994b. Enrichment of bifidobacteria
from human gut contents by oligofructose using continuous
culture. FEMS Microbiol. Lett. 118(1–2): 121–127. doi:10.1111/j.
1574-6968.1994.tb06813.x. PMID:8013867.
Gonzalez, C.F., and Kunka, B.S. 1983. Plasmid transfer in
Pediococcus spp.: intergeneric and intrageneric transfer of
pIP501. Appl. Environ. Microbiol. 46(1): 81–89. PMID:6311098.
Gonzalez, C.F., and Kunka, B.S. 1986. Evidence for plasmid linkage
of raffinose utilization and associated a-galactosidase and sucrose
hydrolase activity in Pediococcus pentosaceus. Appl. Environ.
Microbiol. 51(1): 105–109. PMID:16346958.
Gonzalez, C.F., and Kunka, B.S. 1987. Plasmid-associated bacter-
iocin production and sucrose fermentation in Pediococcus
Can. J. Microbiol. Vol. 57, 2011
acidilactici. Appl. Environ. Microbiol. 53(10): 2534–2538.
PMID:16347470.
González, R., Klaassens, E.S., Malinen, E., de Vos, W.M., and
Vaughan, E.E. 2008. Differential transcriptional response of
Bifidobacterium longum to human milk, formula milk, and
galactooligosaccharide. Appl. Environ. Microbiol. 74(15): 4686–
4694. doi:10.1128/AEM.00122-08. PMID:18539808.
Grimoud, J., Durand, H., Courtin, C., Monsan, P., Ouarne, F.,
Theodorou, V., and Roques, C. 2010. In vitro screening of
probiotic lactic acid bacteria and prebiotic glucooligosaccharides
to select effective synbiotics. Anaerobe, 16(5): 493–500. doi:10.
1016/j.anaerobe.2010.07.005. PMID:20670686.
Grootaert, C., Delcour, J.A., Courtin, C.M., Broekaert, W.F.,
Verstraete, W., and Van de Wiele, T. 2007. Microbial metabolism
and prebiotic potency of arabinoxylan oligosaccharides in the
human intestine. Trends Food Sci. Technol. 18(2): 64–71. doi:10.
1016/j.tifs.2006.08.004.
Gupta, V., and Garg, R. 2009. Probiotics. Int. J. Med. Microbiol. 27(3):
202–209. doi:10.4103/0255-0857.53201.
Hopkins, M.J., Englyst, H.N., Macfarlane, S., Furrie, E., Macfarlane,
G.T., and McBain, A.J. 2003. Degradation of cross-linked and
non-cross-linked arabinoxylans by the intestinal microbiota in
children. Appl. Environ. Microbiol. 69(11): 6354–6360. doi:10.
1128/AEM.69.11.6354-6360.2003. PMID:14602586.
Jaskari, J., Kontula, P., Siitonen, A., Jousimies-Somer, H., Mattila-
Sandholm, T., and Poutanen, K. 1998. Oat b-glucan and xylan
hydrolysates as selective substrates for Bifidobacterium and
Lactobacillus strains. Appl. Microbiol. Biotechnol. 49(2): 175–
181. doi:10.1007/s002530051155. PMID:9534257.
Jung, S.J., Houde, R., Baurhoo, B., Zhao, X., and Lee, B.H. 2008.
Effect of galacto-oligosaccharides and a Bifidobacteria lactis-
based probiotic strain on the growth performance and fecal
microflora of broiler chickens. Poult. Sci. 87(9): 1694–1699.
doi:10.3382/ps.2007-00489. PMID:18753434.
Kaplan, H., and Hutkins, R.W. 2000. Fermentation of fructooligo-
saccharides by lactic acid bacteria and bifidobactria. Appl.
Environ. Microbiol. 66(6): 2682–2684. doi:10.1128/AEM.66.6.
2682-2684.2000. PMID:10831458.
Kedia, G., Vazquez, J.A., and Pandiella, S.S. 2008. Evaluation of the
fermentability of oat fractions obtained by debranning using lactic
acid bacteria. J. Appl. Microbiol. 105(4): 1227–1237. doi:10.1111/
j.1365-2672.2008.03864.x. PMID:18713289.
Kelly, G. 2008. Inulin-type prebiotics — a review: part 1. Altern.
Med. Rev. 13(4): 315–329. PMID:19152479.
Kobayashi, T., Yasutake, N., Uchida, K., Ohyama, W., Kaneko, K.,
and Onoue, M. 2009. Safety of a novel galacto-oligosaccharide:
genotoxicity and repeated oral dose studies. Hum. Exp. Toxicol.
28(10): 619–630. doi:10.1177/0960327109346789. PMID:
19755435.
Lebeer, S., Vanderleyden, J., and de Keersmaecker, S.C.J. 2008.
Genes and molecules of lactobacilli supporting probiotic action.
Microbiol. Mol. Biol. Rev. 72(4): 728–764. doi:10.1128/MMBR.
00017-08. PMID:19052326.
Malinen, E., Matto, J., Salmitie, M., Alander, M., Saarela, M., and
Palva, A. 2002. PCR-ELISA-II: analysis of Bifidobacterium
populations in human faecal samples from a consumption trial
with Bifidobacterium lactis Bb-12 and a galacto-oligosaccharide
preparation. Syst. Appl. Microbiol. 25(2): 249–258. doi:10.1016/
S0723-2020(04)70109-5. PMID:12353880.
Marco, M.L., Pavan, S., and Kleerebezem, M. 2006. Towards
understanding molecular modes of probiotic action. Curr. Opin.
Biotechnol. 17(2): 204–210. doi:10.1016/j.copbio.2006.02.005.
PMID:16510275.
Mlobeli, N.T., Gutierrez, N.A., and Maddox, I.S. 1998. Physiology
Published by NRC Research Press
 Mei et al.
and kinetics of Bifidobacterium bifidum during growth on different
sugars. Appl. Microbiol. Biotechnol. 50(1): 125–128. doi:10.1007/
s002530051266.
Modler, H.W. 1994. Bifidogenic factors-sources, metabolism and
applications. Int. Dairy J. 4(5): 383–407. doi:10.1016/0958-6946
(94)90055-8.
Ng, S.C., Hart, A.L., Kamm, M.A., Stagg, A.J., and Knight, S.C.
2009. Mechanisms of action of probiotics: recent advances.
Inflamm. Bowel Dis. 15(2): 300–310. doi:10.1002/ibd.20602.
PMID:18626975.
Oliveira, R.P.S., Florence, A.C.R., Silva, R.C., Perego, P., Converti,
A., Gioielli, L.A., and Oliveira, M.N. 2009. Effect of different
Can. J. Microbiol. Downloaded from www.nrcresearchpress.com by CONCORDIA UNIV on 03/19/13
prebiotics on the fermentation kinetics, probiotic survival and fatty
acids profiles in non-fat synbiotic fermented milk. Int. J. Food
Microbiol. 128(3): 467–472. doi:10.1016/j.ijfoodmicro.2008.10.
012. PMID:19000641.
Oliveira, R.P.S., Florence, A.C.R., Perego, P., Oliveira, M.N., and
Converti, A. 2011a. Use of lactulose as prebiotic and its influence
on the growth, acidification profile and viable counts of different
probiotics in fermented skim milk. Int. J. Food Microbiol. 145(1):
22–27. doi:10.1016/j.ijfoodmicro.2010.11.011. PMID:21144608.
Oliveira, R.P.S., Perego, P., Oliveira, M.N., and Converti, A. 2011b.
Effect of inulin as a prebiotic to improve growth and counts of a
probiotic cocktail in fermented skim milk. LWT-Food Sci.
Technol. 44(2): 520–523 doi:10.1016/j.lwt.2010.08.024..
Palframan, R.J., Gibson, G.R., and Rastall, R.A. 2002. Effect of pH
and dose on the growth of gut bacteria on prebiotic carbohydrates
For personal use only.
in vitro. Anaerobe, 8(5): 287–292. doi:10.1006/anae.2002.0434.
PMID:16887671.
Palframan, R.J., Gibson, G.R., and Rastall, R.A. 2003. Carbohydrate
preference of Bifidobacterium species isolated from the human
gut. Curr. Issues Intest. Microbiol. 4(2): 71–75. PMID:14503691.
Perrin, S., Fougnies, C., Grill, J.P., Jacobs, H., and Schneider, F.
2002. Fermentation of chicory fructo-oligosaccharides in mixtures
of different degrees of polymerization by three strains of
bifidobacteria. Can. J. Microbiol. 48(8): 759–763. doi:10.1139/
w02-065. PMID:12381033.
Pylkas, A.M., Juneja, L.R., and Slavin, J.L. 2005. Comparison of
different fibres for in vitro production of short chain fatty acids by
intestinal microflora. J. Med. Food, 8(1): 113–116. doi:10.1089/
jmf.2005.8.113. PMID:15857221.
Rabiu, B.A., Jay, A.J., Gibson, G.R., and Rastall, R.A. 2001.
Synthesis and fermentation properties of novel galacto-oligosac-
charides by b-galactosidases from Bifidobacterium species. Appl.
Environ. Microbiol. 67(6): 2526–2530. doi:10.1128/AEM.67.6.
2526-2530.2001. PMID:11375159.
Raccach, M. 1987. Pediococci and biotechnology. Crit. Rev.
Microbiol. 14(4): 291–309. doi:10.3109/10408418709104442.
PMID:3308317.
Roberfroid, M. 2007. Prebiotics: the concept revisited. J. Nutr. 137(3
Suppl. 2): 830S–837S. PMID:17311983.
Roberfroid, M.B., Van Loo, J.A., and Gibson, G.R. 1998. The
865
bifidogenic nature of chicory inulin and its hydrolysis products. J.
Nutr. 128(1): 11–19. PMID:9430596.
Ryan, K.A., Daly, P., Li, Y., Hooton, C., and O’Toole, P.W. 2008.
Strain-specific inhibition of Helicobacter pylori by Lactobacillus
salivarius and other lactobacilli. J. Antimicrob. Chemother. 61(4):
831–834. doi:10.1093/jac/dkn040. PMID:18263567.
Rycroft, C.E., Jones, M.R., Gibson, G.R., and Rastall, R.A. 2001. A
comparative in vitro evaluation of the fermentation properties or
prebiotic oligosaccharides. J. Appl. Microbiol. 91(5): 878–887.
doi:10.1046/j.1365-2672.2001.01446.x. PMID:11722666.
Sako, T., Matsumoto, K., and Tanaka, R. 1999. Recent progress on
research and applications of non-digestible galacto-oligosacchar-
ides. Int. Dairy J. 9(1): 69–80. doi:10.1016/S0958-6946(99)
00046-1.
Sanz, M.L., Gibson, G.R., and Rastall, R.A. 2005. Influence of
disaccharide structure on prebiotic selectivity in vitro. J. Agric.
Food Chem. 53(13): 5192–5199. doi:10.1021/jf050276w. PMID:
15969496.
Snart, J., Bibiloni, R., Grayson, T., Lay, C., Zhang, H., Allison, G.E.,
et al. 2006. Supplementation of the diet with high-viscosity b-glucan
results in enrichment for lactobacilli in the rat cecum. Appl.
Environ. Microbiol. 72(3): 1925–1931. doi:10.1128/AEM.72.3.
1925-1931.2006. PMID:16517639.
Tannock, G.W., Munro, K., Bibiloni, R., Simon, M.A., Hargreaves,
P., Gopal, P., et al. 2004. Impact of consumption of oligosacchar-
ide-containing biscuits on the fecal microbiota of humans. Appl.
Environ. Microbiol. 70(4): 2129–2136. doi:10.1128/AEM.70.4.
2129-2136.2004. PMID:15066805.
Tzortzis, G., Goulas, A.K., Gee, J.M., and Gibson, G.R. 2005. A novel
galactooligosaccharide mixture increases the bifidobacterial popu-
lation numbers in a continuous in vitro fermentation system and in
the proximal colonic contents of pigs in vivo. J. Nutr. 135(7):
1726–1731. PMID:15987856.
Van Craeyveld, V., Swennen, K., Dornez, E., Van de Wiele, T.,
Marzorati, M., Verstraete, W., et al. 2008. Structurally different
wheat-derived arabinoxylooligosaccharides have different prebiot-
ic and fermentation properties in rats. J. Nutr. 138(12): 2348–2355.
doi:10.3945/jn.108.094367. PMID:19022956.
Van der Meulen, R., Avonts, L., and De Vuyst, L. 2004. Short
fractions of oligofructose are preferentially metabolized by
Bifidobacterium animalis DN-173 010. Appl. Environ. Microbiol.
70(4): 1923–1930. doi:10.1128/AEM.70.4.1923-1930.2004.
PMID:15066781.
Van der Meulen, R.V., Makras, L., Verbrugghe, K., Adriany, T., and De
Vuyst, L.D. 2006. In vitro kinetic analysis of oligofructose
consumption by Bacteroides and Bifidobacterium spp. indicates
different degradation mechanisms. Appl. Environ. Microbiol. 72(2):
1006–1012. doi:10.1128/AEM.72.2.1006-1012.2006. PMID:
16461642.
Vernazza, C.L., Gibson, G.R., and Rastall, R.A. 2006. Carbohydrate
preference, acid tolerance and bile tolerance in five strains of
Bifidobacterium. J. Appl. Microbiol. 100(4): 846–853. doi:10.
1111/j.1365-2672.2006.02832.x. PMID:16553741.
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