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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,
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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
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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- water and 5% filter-sterilized L-cysteine HCl monohydrate
cillus strains and 7 of 8 Bifidobacterium strains tested were (FisherBiotech)) for 24 h. L-Cysteine was added to provide
able to ferment the substrates. Several studies have demon- amino nitrogen as a growth factor and to reduce redox poten-
strated that inulin and oligofructose are selectively fermented tial. Bifidobacterium strains were grown in Reinforced Clos-
by bifidobacteria (Roberfroid et al. 1998; Perrin et al. 2002; tridial Medium broth (Oxoid) at 37 °C for 48 h under
Oliveira et al. 2009, 2011b). Jaskari et al. (1998) reported anaerobic atmosphere using GasPak EZ Anaerobe Container
that b-glucooligomers (from oat) and raffinose enhance the System Sachets (Becton Dickinson and Company, Maryland,
growth of health-promoting probiotic strains but only raffi- USA) in the GasPak EZ Incubation Container (Becton Dick-
nose does it at a significant level. In addition, a comparative inson and Company). Dry anaerobic indicators strips (Becton
study showed that GOS were effective at increasing the num- Dickinson and Company) were used to verify that an anaero-
ber of bifidobacteria and lactate while generating less gas bic atmosphere was attained.
(Rycroft et al. 2001). It has been also shown that oral admin-
istration of GOS enhances faecal levels of Bifidobacterium Prebiotics
animalis subsp. lactis (Malinen et al. 2002). Kedia et al.
Sixteen different types of dietary fibers were examined.
(2008) evaluated the fermentability of oat fractions obtained
The carbohydrates, degree of polymerization, and where ap-
by debranning using three lactobacilli and demonstrated that
propriate, the commercial source are shown in Table 2. Glu-
a 1%–3% pearling fraction, containing the highest concentra-
cose and galactose, used as controls, were purchased from
tion of total dietary fibre compared with other pearling frac-
Sigma.
tions, is the most suitable for fermentation and produces
considerably higher probiotics cell concentrations.
The objective of this study was to examine the utilization Basal medium used for prebiotics studies
of a number of plant oligosaccharides and polysaccharides as The basal medium (Degnan and Macfarlane 1995) was
prebiotics (which are characterized by different linear chain used as a carbohydrate-free medium for testing the effect of
components) by selected probiotic bacteria. This will provide the potential prebiotics on the growth of the probiotic strains.
important information about the factors that influence the fer- The basal medium consisted of (g/L) peptone water, 5.0;
mentation of nondigestible carbohydrates by the probiotic tryptone, 10; yeast extract, 2.5; Tween 80, 1.0; NaCl, 4.5;
bacteria and information that can be used to determine which KCl, 0.25; MgCl2·6H2O, 0.15; KH2PO4, 0.40; K2HPO4,
types of prebiotics are suitable to prepare new and improved 0.20; NH4Cl, 0.40; and cysteine–HCl, 0.50.
prebiotic–probiotic formulations.
Preparation of cell suspensions
Materials and methods In preparation for the fermentation trials, the bacteria were
grown in 10 mL of the appropriate medium (modified
Bacterial strains and growth conditions de Mann – Rogosa – Sharpe or Reinforced Clostridial Me-
The bacterial strains used in this study are listed in Table 1. dium) containing glucose (10 g/L) as the carbon source.
Lactobacillus spp. and Pediococcus spp. were grown in a 5% 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.
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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 cance between treatment and control conditions was assessed
physiological saline (0.85% NaCl solution), and resuspended by the Tukey’s test. A significant difference was defined as a
in 10 mL of basal medium to remove excess carbon prior to P value of <0.05.
the fermentation trials.
Results
In vitro fermentation
Growth of the probiotic strains was monitored in basal me- FOS from rye and chicory greatly promoted the growth of
dium supplemented with 1% of each test prebiotic as the sole all the strains tested (Fig. 1). Consequently, the bacterial me-
carbon source (Table 2). Following media preparation, a 1% tabolism of these substrates caused a marked decrease in the
prewashed inoculum of a given probiotic culture was used to culture medium pH (Fig. 1). All the Bifidobacterium strains
examine the effect of the selected prebiotics on the growth of tested and P. pentosaceus FRP 243 also showed a great abil-
the probiotic culture. Cultures (10 mL) were incubated at ity to utilize inulin from chicory root (Figs. 1B and 1C).
37 °C for a period of 48 h under anaerobic conditions. Each No influence of b-glucan (from either oat or barley) or
fermentation experiment was performed in triplicate. The other polysaccharides on the growth of probiotics or the pH
controls consisted of inoculated basal medium lacking prebiotics of culture medium was observed (Fig. 2); whereas, all the pro-
and inoculated basal medium containing glucose or galactose. biotics tested grew very well in b-glucotriose (3-o-b-glucosyl-
The growth of each strain was monitored at stationary cul- D-cellobiose prepared by enzymatic hydrolysis of barley b-
ture phase at 48 h by measuring the optical density (OD) of glucan using lichenase (Megazyme)), and the corresponding
the cultures at 600 nm using a spectrophotometer (Ultrospec pH of the medium from fermentation significantly decreased
3100 pro). (P < 0.05) (Fig. 3). With the exception of Lactobacillus
The pH of aliquots was also determined at stationary cul- curvatus (FRP 15) and P. acidilactici (FRP 236), all other
ture phase (48 h) using a pH meter (Fisher Scientific), soon strains were able to utilize and ferment b-glucotetrose (3-o-
after removing the samples from the fermentation vessels. b-glucosyl-D-cellotriose) prepared by enzymatic hydrolysis
of barley b-glucan (Megazyme). Lactobacillus rhamnosus
Data and statistical analysis GG (ATCC 53103), L. kefir IM002, B. breve (FRP 334),
For each experiment, the data were analyzed using the Ex- and P. pentosaceus (FRP 244) grew well in b-glucopentose
cel statistical package. The OD and pH readings and corre- (3-o-b-cellotetraosyl-D-glucose) (Megazyme). Bifidobacterium
sponding standard deviations were calculated from triplicate breve (FRP 334) was the only probiotic strain that demon-
samples from three separate experiments. Statistical signifi- 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
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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.
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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 Can. J. Microbiol. Vol. 57, 2011

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