Physiological Changes of Surface Membrane

in Lactobacillus with Prebiotics

Mingfang Pan, Kishore K. Kumaree, and Nagendra P. Shah

Abstract: Synbiotics are always considered to be beneficial in healthy manipulation of gut environment; however, the
purpose of this research was to investigate the dominance of synbiotic over the individual potential of probiotics and
prebiotics. Four different types of prebiotics, fructo-oligosaccharides, raffinose, inulin, and cellobiose, were evaluated
based on their varying degree of polymerization, combined each with 2 different Lactobacilli strains, including Lactobacillus
paracasei 276 and Lactobacillus plantarum WCFS1. The effects of synbiotics combination on the surface structure were
evaluated by analyzing auto-aggregation, membrane hydrophobicity, and adhesion to Caco-2 cells. Our results showed
that both Lactobacilli exhibited significantly greater degree of attachment to Caco-2 cells (23.31% and 16.85%, respectively)
when using cellobiose as a substrate than with other prebiotics (P < 0.05). Intestinal adhesion ability was in correlation with
the percent of auto-aggregation, both Lactobacillus exhibited higher percent of auto-aggregation in cellobiose compared
to other prebiotics. These behavioral changes in terms of attachment and auto-aggregation were further supported with
the changes noticed from infrared spectra (FT-IR).

Keywords: Lactobacillus, membrane hydrophobicity, prebiotics, probiotics

Introduction to intestinal wall and eventually promoting healthy colonization

The term probiotics is defined as “living microorganisms, which
when administered in adequate amounts, confer a health benefit
on the host” (FAO/WHO 2002). Adhesion to intestinal wall is
one of the important characteristics of a bacterium for considering
it as a probiotic, as its attachment to the human intestinal wall can
increase its efficacy. Adhesion is also essential for the establishment
and initiation of healthy colonization. Auto-aggregation is another
important characteristic for probiotics; this property helps them
Food Microbiology &
(Kankaanpaa and others 2001; Schar-Zammaretti and others 2005;
Deepika and others 2009). Studies have been carried out to explore
the influence of various established media on the microbial sur-
face properties by using various physicochemical methods (Schar-
Zammaretti and other 2005). However, the effects of different
prebiotics on the physicochemical properties of probiotics have
not been studies fully. Prebiotics are the functional compounds
that have the potential to influence the physiology of whole body
directly or indirectly (Gibson and Roberfroid 1995; Aachary and

attach to the intestinal mucosa. Moreover, the predominance of

these organisms in the gut can prevent the growth of unwanted Prapulla 2011). A combination of probiotic(s) and prebiotic(s)
pathogens (Schachtsiek and others 2004). constitutes a synbiotic (Gourbeyre and others 2011), which can
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Attachment to epithelial wall is an intricate process involving
nonspecific and specific ligand-receptor mechanisms. The adher-
ence of bacteria is initially linked with the surface properties which
trigger the physicochemical interactions between 2 surfaces. Past
research (Garcı́a-Cayuela and others 2014) on Lactobacillus has cor-
related adhesion ability with hydrophobicity, as measured by mi-
crobial affinity to hydrocarbons. However, contradictory results are
reported by Vinderola and others (2004). Additionally, physico-
chemical characteristics of the cell surface, such as hydrophobicity,
have been reported to affect aggregation abilities (Kos and oth-
ers 2003). Previous work indicates that the surface proteins of
some Lactobacillus contribute to the membrane hydrophobicity,
thus helping the adherence of Lactobacillus to the gut wall (Mukai
and Arihara 1994). As a result, adhesion, surface hydrophobicity,
auto-aggregation, and co-aggregation are phenotypic traits that
provide potential microbial colonization advantages within the
intestinal tract.
There are several physicochemical properties that affect mem-
brane characteristics, which help the adherence of Lactobacillus
JFDS-2016-1677 Submitted 10/12/2016, Accepted 12/9/2016. All authors
are with Food and Nutritional Science, School of Biological Sciences, The Univ. of
Hong Kong, Pokfulam Road, Hong Kong. Author Shah is also the Adjunct Profes-
sor, Victoria Univ., Melbourne, Australia. Direct inquiries to author Shah (E-mail:
npshah@hku.hk).
stimulate the growth of probiotics and may increase the survival
of these organisms in the intestinal tract. Providing prebiotics as
substrates to probiotics may increase the surface hydrophobicity or
attachment potential.
Fourier transform infrared analysis (FT-IR) can be utilized for
in situ analysis of surface functional groups of bacteria. It has been
used to analyze microorganisms for the last 4 decades. Unidentified
microorganisms can be recognized by the comparative analysis
of their FT-IR spectra (Oust and others 2004). Proteins, lipids,
and sugars in the bacterial membranes have distinguished infrared
(IR) vibrations that indicate their conformation and physical state.
In this aspect, FT-IR analysis has proven to be a powerful tool
to understand changes in biological membranes as affected by
variation in a medium added with prebiotics (Oldenhof and others
2005).
Earlier studies on the beneficial effects of synbiotics are based
on clinical trials (Olah and others 2002; Bengmark 2005) and
have not examined in-depth information behind the synbiotic ef-
fects at the bacterial cell level. Hence, in this study we aimed
at evaluating whether bacterial aggregation abilities, cell sur-
face hydrophobicity, and adhesion abilities of 2 strains of Lac-
tobacillus are affected by different prebiotics. FT-IR analysis was
used to examine the membrane behavior of bacterial cells and
changes in the vibrational pattern of surface functional groups in
them.

C 2017 Institute of Food Technologists

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744 Journal of Food Science r Vol. 82, Nr. 3, 2017 doi: 10.1111/1750-3841.13608
Further reproduction without permission is prohibited
Potential of probiotics and prebiotics . . .

Table 1–Auto-aggregation abilities of the 2 Lactobacillus strains determined spectrophotometrically.

Aggregation (%)
Synbiotic combination
Strains (mMRS+ prebiotic) 4h 24 h
L. paracasei 276 FOS 45.4 ± 2.04c 63.1 ± 2.50b
Inulin 47.6 ± 1.3b/c 65.8 ± 0.75b
Raffinose 51.8 ± 0.98a/b 63.8 ± 1.74b
Cellobiose 54.0 ± 0.19a 77.1 ± 1.69a
L. plantarum WSFC-1 FOS 44.1 ± 0.74c 67.9 ± 0.25c
Inulin 53.9 ± 0.27b 70.9 ± 0.63b
Raffinose 53.8 ± 0.05b 66.4 ± 0.15d
Cellobiose 70.3 ± 0.63a 81.4 ± 0.17a
Values are means ± SEM of experiment conducted in duplicates.
Mean values for each strain at each incubation time with no common superscript letters differ significantly, P < 0.05.

Materials and Methods was measured at 600 nm (Multiskan GO plate reader) at 4 h and
24 h. The percent of auto-aggregation was calculated as a function
Bacterial strains of time using the following equation: [1- (At / A0 )] × 100, here At
Two strains of lactic acid bacteria, Lactobacillus paracasei 276 and is the absorbance recorded at 600 nm at the defined time period
L. plantarum WCFS 1, were previously obtained from Australian (that is, 4 h and 24 h) and A0 is at the initial value.
Starter culture Collection and stored at –80. After activation, these
bacteria were grown in modified MRS (composition: casein pep-
tone, tryptic digest 10 g/L; meat extract 10 g/L; Tween 80 1 g/L; Bacterial adhesion to hydrocarbons
K2HPO4 2 g/L; Na-acetate 5 g/L; MgSO4.7H2O 0.20 g/L) Bacterial membrane hydrophobicity or hydrophilic nature of
supplemented with 2% of selected prebiotics including: raftilose the cell surface was determined by their affinity to hydrocarbon.
P95-FOS and raftilose HP-inulin (Orafti, Active Food Ingredients, Bacterial adhesion to hydrocarbon was performed as previously
Tienen, Belgium), Cellobiose and Raffionse (Sigma Aldrich, Saint described (Garcı́a-Cayuela and others 2014) with some modifi-
Louis, Mo., U.S.A.). cations. Lactobacillus grown overnight in mMRS was harvested,
washed 2 times with phosphate potassium urea and magnesium
(PUM buffer: pH 7.1; 22.2 g K2 HPO4 •3H2 O, 7.26 g KH2 PO4 ,
Auto-aggregation assay 1.8 g urea, 0.2 g MgSO4 •7H2 0 and distilled water to 1000 mL)
Auto-aggregation test was carried out as previously described and resuspended in the same buffer to maintain an initial OD
(Kotzamanidis and others 2010) with some modifications. The (ODI -600) range of 350 to 400 at 600 nm. The initial OD was
organisms were grown in modified MRS (mMRS) under micro standardized to maintain an equal number of bacterial cells. An

Food Microbiology &

aerophilic condition for 24 h, cells were harvested postincuba- aliquot of the bacterial cells (1 mL) was mixed with 0.4 mL of xy-
tion and resuspended with the same volume of phosphate buffer lene (apolar solvent) and the solution was vortexed vigorously for

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(PBS; Na2 HPO4 -NaH2 PO4 , pH 7.2). The suspension was vor- 120 s. The dual phase mixture was left at 37 °C to ensure proper
texed thoroughly and the optical density (OD) of the upper layer separation of aqueous phase and organic phase. After 30 min

Figure 1–The hydrophobicity (%) of 2 Lactobacillus strains after 24 h of incubation. Data are means and standard errors of 3 independent assays.
Columns for each strain that do not share the same letter are significantly different (P < 0.05).

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 Potential of probiotics and prebiotics . . .

Table 2–Correlation matrix of Pearson coefficients between aggregation, hydrophobicity, and adhesion properties.

Auto-aggregation Hydrophobicity Caco-2 cell adhesion

L. paracasei 276
Auto-aggregation −
Hydrophobicity 0.271 −
Caco-2 cell adhesion 0.950∗∗ 0.300 −
L. plantarum WCFS-1
Auto-aggregation −
Hydrophobicity 0.859 −
Caco-2 cell adhesion 0.616 0.201 −
The correlation coefficient measures the degree of linear dependence between each pair of variables.
∗∗
Correlation is significant at P < 0.01 level.

of incubation, the bottom layer of aqueous phase was carefully
separated and the final absorbance was recorded (ODF -600). The
comparison of initial (ODI -600) with the final absorbance (ODF -
600) gave the affinity of bacteria for hydrocarbon and loss from
the aqueous phase. The following formula was used to determine
the percent hydrophobicity of the membrane:
%Hydrophobicity = [(ODinitial −ODfinal ) /ODinitial ] ×100
Adhesion Assay
Cell culture
The adhesive property of Lactobacillus in different prebiotics was
assayed using colonocyte-like cell lines, Caco-2 (American Type
Culture Collection, Manassas, Va., U.S.A.). The culture of cell line
was carried out as mentioned previously (Gandhi and Shah 2016)
with the following modification. Caco-2 cells were cultured in
minimum essential medium eagle (EMEM; Sigma-Aldrich) sup-
plemented with 10% fetal bovine serum (FBS) and 1% penicillin-
streptomycin solution (Gibco BRL, Grand Island, N.Y., U.S.A.).
The culture was maintained through incubation at 37 °C with
Food Microbiology &
under strict aseptic condition. Passaging and seeding of Caco-2
cells were done at 80% confluence with proper monitoring of
their growth.
Assay of adhesion to intestinal epithelial-like cell line
Caco-2 cells were seeded (1 ×104 cell/mL) in 96-well tissue
culture plates (NUNC, Thermofisher Scientific, Waltham, Mass.,
U.S.A.). Plates were incubated under 95% RH at 37 °C with 5%
CO2 supply to attain 80% confluence prior to the adhesion assay.
Overnight bacterial cells grown in mMRS were harvested, washed
twice with PBS (pH 7.2), and resuspended in EMEM medium
containing 10% of FBS without any antibiotic. The initial con-
centration of each combination was maintained at approximately
108 CFU/mL. In the meantime, Caco-2 monolayers were also
washed twice with PBS to eliminate the traces of antibiotics and
incubated with live bacteria separately and EMEM solution (10:1;
bacterium: eukaryotic cells). The plates were incubated for 120
min at 37 °C and 5% CO2 . After incubation, the supernatant
was discarded and the wells were gently washed with PBS. The at-
tached bacteria were removed from the monolayers by trypsinizing

a continuous supply of 5% CO2 and 95% RH. The viability with 0.25 % of trypsin-EDTA solution (Gibco BRL) and plating
of the cells was assured by the regular supply of fresh medium the appropriate dilution on MRS plates. The number of bacteria
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Figure 2–Percent adhesion (bacteria adhered with respect to the number of bacteria added) of the 2 Lactobacillus strains to the intestinal Caco-2 cell
line.
Each adhesion assay was conducted in triplicate with cells from 3 successive passages.
Means and standard errors are shown.
Columns for each strain that do not share the same letter are significantly different (P < 0.05).

746 Journal of Food Science r Vol. 82, Nr. 3, 2017

Potential of probiotics and prebiotics . . .

attached to the cell line was expressed as the percent adhesion with

hydrocarbons
the following formula:

aromatic
=CH in

891.67
893.86
892.64
894.67
893.61
893.61
890.64
889.97
Table 3–Absorption wavenumbers observed for subjected to varying synbiotic combinations yielded closer FT-IR spectra for L. paracasei 276 and L. plantarum WCFS-1. %Adhesion = [(attachedCFU/initialCFU) ×100]

Sample preparation for FT-IR

The samples for FT-IR analysis were prepared as described ear-
lier (Gandhi and Shah 2014). Bacterial cells exposed to different
C–H def of prebiotics containing mMRS medium were centrifuged separately
>CH2 of
Amide section of protein

proteins
1455.42
1452.72
1454.52

1453.91
1451.82
1454.67
1456.01

1453.33
at 5000 × g for 15 min at 4 °C and resuspended in PBS buffer
(100 mg: 0.25 mL buffer). An aliquot of 50 µL of bacterial sample
was placed at the center of the CaF2 plates and oven dried (60 °C
Amide II and II’

for 20 min) in order to get a layer of solid film. The measurements

region of

were carried out with FT-IR spectrophotometer (Perkin Elmer,

protein

Waltham, Mass., U.S.A.; Spectrum Two, LiTa03 detector, MIR

Amide II
1554.37
1553.37
1553.92
1551.67
1553.11
1555.37
1553.11
1553.11
source and essential) equipped with DTGS detector. The measure-
ments were recorded with the wave number range of 4000 cm−1
to 400 cm−1 . A total of 20 scans were recorded for each sample
with the resolution of 4 cm−1 . The reference spectrum (back-
1648.37
1647.47
1647.92

1648.9
1646.57
1648.61
1647.88

1647.99

ground) was obtained by running IR in an empty chamber.

Amide I & I’ of

(C O stretching
of amide)
structure

Statistical analysis
α helical

Mean values, standard errors, and correlation coefficients were

1660.96
1660.96
1661.86
1664.11
1661.43
1661.26
1661.59
1661.43

evaluated for the values of the different variables studied. One-

way ANOVA was used to determine the significant difference
(P < 0.05, Duncan test) for the auto-aggregation, hydrophobicity,
and adhesion among different prebiotics treatments. Coefficient
of correlation analysis was performed by calculating the Pearson
>CH2 in fatty
stretching of
symmetric

product–moment correlation and all data obtained were calculated

2851.97

2853.76
2852.43
2859.41
2858.41
2852
2852
2852
-C-H

acids

by analysis of variance (ANOVA) using SPSS (Li and others 2010).

Results and Discussion

Auto-aggregation

Food Microbiology &

Spectrophotometric detection of auto-aggregation of the 2 Lac-
>CH2 in fatty
stretching of
asymmetric

tobacillus strains in media containing different prebiotics is pre-

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2925.56
2924.04

2924.92
2922.92
2928.24
2927.91
2926.5
2928.5
acids

sented in Table 1. Incubation times of 4 and 24 h were chosen to

C-H

monitor the change in auto-aggregation. As shown in the table,

the percent aggregation increased with the duration of incubation.
After 4-h incubation, the highest aggregation of L. paracasei 276
was observed in the case of cellobiose followed by insulin, FOS,
and raffinose. When incubation time extended to 24 h, the highest
-CH3 in fatty
stretching of
asymmetric

value was still found in cellobiose (77.1%, but the percent aggre-
Sadiq and others (2015), Gandhi and Shah (2014), Martins and others (2010).

gation in other 3 prebiotics was all around 65%. A similar pattern

acids
C-H

2960
2960
2960

2960
2960
2960
2960

2960

was also observed for L. plantarum WCFS1, after 24 h incubation,

the highest aggregation was observed in cellobiose (81.4 ± 0.17 %)
when compared to other 3 prebiotics. Aggregation has been linked
to the adhesion of Lactobacillus in a previous study (Garcı́a-Cayuela
and others 2014). Another work by Del and others (2000) on Bifi-
Prebiotic sugar

dobacterium also demonstrated the relationship of auto-aggregation

with adhesion ability. In that work, Bifidobacterium which showed
Cellobiose

Cellobiose
Raffinose

Raffinose

higher attachment to epithelial cells also had higher aggregation

Inulin

Inulin

ability among other 13 B. longum species.

FOS

FOS

Membrane hydrophobicity
Bacterial membrane hydrophobicity is determined by its adhe-
L. plantarum WCFS-1
Lactobacillus strains

sion to hydrocarbons. In this study, varying adhesion to xylene for

Lactobacillus grown in medium containing different prebiotics was
L. paracasei 276

measured (Figure 1). The percent hydrophobicity for L. paracasei

276 was the highest in the medium with FOS (77.52 ± 0.3%)
although it showed no significant difference (P > 0.05) with cel-
lobiose (75.62 ± 3.02%). In raffinose, however, the bacterium

Vol. 82, Nr. 3, 2017 r Journal of Food Science 747

 Potential of probiotics and prebiotics . . .
showed the lowest percent of hydrophobicity. A similar trend was
observed in the case of L. plantarum WCFS-1. This organism
grown in the medium with cellobiose exhibited relatively higher
percentage of hydrophobicity (81.93 ± 0.79%) when compared
with others. While in the raffinose, it showed the lowest values as
well. The result is in accordance with the earlier work by Li and
others (2010), the carbohydrate source (sucrose) in the growth
medium affected the surface parameters henceforth influencing
membrane hydrophobicity of the bacterium, L. casei Zhang.
Adhesion to Caco-2 cells
Adhesion is considered as an important characteristic for pro-
biotic bacteria in order to establish colonization in the intestine.
In this study, the adhesion ability of Lactobacillus was examined
to understand the influence of different prebiotics on adhesion to
Caco-2 cell (Figure 2). For L. paracasei 276, the highest adhesion
was observed in cellobiose (23.31 ± 0.64 %) followed by inulin
(13.59 ± 0.53 %). However, the adhesion in the medium with
FOS or raffinose showed the lowest values at 5.9% and 6.18%,
Food Microbiology &
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748 Journal of Food Science r Vol. 82, Nr. 3, 2017
respectively. Similarly, the greatest degree of adhesion for L. plan-
tarum WCFS-1 was observed in the medium containing cellobiose
as well (16.85 ± 1.20 %). However, the performance of adhesion
in the other 3 prebiotics was different, with FOS and raffinose
ranked at 2nd place (10.73% and 10.97%, respectively) and inulin
had the lowest values.
For L. paracasei 276, the presence of cellobiose not only showed
the highest level of adhesion but also contributed a high degree of
auto-aggregation and relatively high membrane hydrophobicity
(Table 1; Figure 1). On the other hand, L. paracasei 276 showed the
highest degree of hydrophobicity in FOS but exhibited the lowest
level of attachment. This discrepancy could be due to other factors
such as membrane electrostatic interaction, and steric forces,
which may have influenced the hydrophobicity of probiotics. For
L. plantarum WCFS-1, the highest level of attachment in cellobiose
also exhibited higher hydrophobicity and auto-aggregation. Sev-
eral studies have shown a relationship between aggregation ability,
membrane hydrophobicity, and adhesion property (Collado
and others 2008; Kotzamanidis and others 2010). Pearson’s
Figure 3–Infrared spectral shifts focusing amide I and
amide II regions of bacterial surface membrane in medium
containing different synbiotic (A) L. paracasei 276 and (B)
L. plantarum WCFS-1.
Potential of probiotics and prebiotics . . .
correlation coefficients between bacterial auto-aggregation
abilities, hydrophobicity, and cell adhesion property are shown in
Table 2. In regards to correlation coefficient for L. paracasei 276,
membrane hydrophobicity exhibited a weak correlation with
auto-aggregation and Caco-2 cell adhesion, whereas there was a
very strong correlation between auto-aggregation and Caco-2 ad-
hesion. In L. plantarum WCFS-1, highest degree of correlation was
observed for aggregation and hydrophobicity, meanwhile adhesion
ability showed medium ranged correlation with auto-aggregation,
whereas hydrophobicity and Caco-2 adhesion exhibited weaker
correlation coefficient. For the 2 strains of Lactobacillus studied, hy-
drophobicity had weak correlation with the adhesion ability; simi-
lar reports were also earlier reported by Garcı́a-Cayuela and others
(2014). The authors found the Lactobacillus plantarum exhibiting
higher adhesion to epithelial wall had lowest hydrophobicity com-
pared to other Lactobacillus strains. In several studies, hydrophobic-
ity has been shown to be correlated with adhesion ability (Del Re
and others 2000), whereas in our study, adhesion ability of both
Lactobacillus strains was not in correlation with hydrophobicity of
membrane. As adhesion ability is a more complex assay involving
several other parameters (surface exo-polysaccharides, S-layer
protein, lipoteichoic acid) and hydrophobicity can also contribute
to adhesion ability to some extent. Interestingly, auto-aggregation
for both strains of Lactobacillus was positively correlated with the
adhesion ability, which was similar to that reported in the earlier
reports (Del and others 2000; Wang and others 2010).
FT-IR spectra analysis of Lactobacillus
Infrared spectra were recorded for the bacterial cells membrane
grown in the medium containing varying prebiotics (Table 3).
The shifts of wavenumbers that occurred at Amide I and Amide
II regions are shown in Figure 3. The dendogram obtained using
Pearson coefficient as the method of generation is presented in
Figure 4. The data from infrared spectrum clearly show that there
was a variation in vibration or stretching of bacterial membrane
functional groups. For L. paracasei 276, there was a close relation-
ship between the spectral bands when the bacterium was grown
in FOS, raffinose, and inulin, whereas the bands in cellobiose was
distinctly separated from others (Figure 4A). However, in the case
of L. plantarum WCFS-1, spectral bands in raffinose and cellobiose
synbiotic were clustered into one group, while the bands in FOS
Table 4–Infrared band absorption ratio of amide (1400 to 1700
cm−1 ) to sugar (800 to 1200 cm−1 ) of L. paracasei 276 and L.
plantarum WCFS-1.
Amide region area / sugar region area
L. paracasei 276 L. plantarum WCFS-1
FOS 0.062 0.047
Inulin 0.084 0.054
Raffinose 0.086 0.113
Cellobiose 0.161 0.110
and inulin formed another group (Figure 4B). The spectra ob-
served from FT-IR are indicative of the changes that occurred in
the surfaces of both Lactobacilli.
Absorption bands at 2960 cm−1 , corresponding to asymmetric
stretching of -CH3 in fatty acids, had no shift in both Lactobacillus
(Table 3). The band between 1700 and 1470 cm−1 referred to
the amide regions of membrane protein structure. The C = O
stretching of amide is referred as amide I and observed at 1600 to
1700 cm−1 (Santivarangkna and others 2007) while N-H bending
at approximately 1554 cm−1 is referred as amide II. In all spectra,
shifts around 1450 to 1450 cm−1 were observed; these are the
characteristic of the scissoring movement of –CH2 groups. Major
significant shifts were observed at amide II region of L. paracasei 276
(Figure 3A) whereas for L. plantarum WCFS-1 (Figure 3B) minor
shifts were observed in the amide region. As reported by Naumann
and others (1982), the cell wall and cell membrane region of 1200
to 800 cm−1 were constantly attributed by the peptidoglycan of
bacteria. Thus alteration in these area spectra may indicate changes
in the surface structure. Changes in membrane caused by different
growth media can also be explained by comparing the areas under
specific infrared spectral regions. For L. paracasei 276, cellobiose
had higher amide area/ sugar area ratio, indicating the amount
Food Microbiology &
of detectable protein was more than the bacterial surface sugar
(Table 4). In case of L. plantarum WCFS-1, raffinose and cellobiose
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had higher ratio of amide area to sugar area (0.113 and 0.11,
respectively). Proteins on the surface of bacteria are an important
parameter for determining their adhesive nature to the epithelial
wall. As reported by Wang and others (2010), higher amount of
surface protein contributes to the membrane hydrophobicity. As
reported earlier, adherence of Lactobacilllus was affected due to the
Figure 4–Dendogram obtained from the absorption
wavenumbers observed for subjected to varying synbiotic
combinations yielded closer FT-IR spectra for (A) L.
paracasei 276 and (B) L. plantarum WCFS-1. With cluster
analysis performed with the Pearson correlation coefficient
and by the Ward’s algorithm method.
Vol. 82, Nr. 3, 2017 r Journal of Food Science 749
 Potential of probiotics and prebiotics . . .
inactivation of surface protein by trypsin (Coconnier and others
1992).
Conclusion
The current work found that different prebiotics had varying
effect on surface characteristics like membrane hydrophobicity,
auto-aggregation, and Caco-2 cell adhesion ability. A further as-
sociation was established by Pearson’s correlation coefficient which
was supported by the FT-IR data. Both Lactobacillus strains showed
greater adhesion ability in the presence of cellobiose, and in-
teresting results from FT-IR also exhibited changes associated
with surface characteristics of Lactobacillus in the presence of this
prebiotic.
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