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

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 stimulate the growth of probiotics and may increase the survival
nonspecific and specific ligand-receptor mechanisms. The adher- of these organisms in the intestinal tract. Providing prebiotics as
ence of bacteria is initially linked with the surface properties which substrates to probiotics may increase the surface hydrophobicity or
trigger the physicochemical interactions between 2 surfaces. Past attachment potential.
research (Garcı́a-Cayuela and others 2014) on Lactobacillus has cor- Fourier transform infrared analysis (FT-IR) can be utilized for
related adhesion ability with hydrophobicity, as measured by mi- in situ analysis of surface functional groups of bacteria. It has been
crobial affinity to hydrocarbons. However, contradictory results are used to analyze microorganisms for the last 4 decades. Unidentified
reported by Vinderola and others (2004). Additionally, physico- microorganisms can be recognized by the comparative analysis
chemical characteristics of the cell surface, such as hydrophobicity, of their FT-IR spectra (Oust and others 2004). Proteins, lipids,
have been reported to affect aggregation abilities (Kos and oth- and sugars in the bacterial membranes have distinguished infrared
ers 2003). Previous work indicates that the surface proteins of (IR) vibrations that indicate their conformation and physical state.
some Lactobacillus contribute to the membrane hydrophobicity, In this aspect, FT-IR analysis has proven to be a powerful tool
thus helping the adherence of Lactobacillus to the gut wall (Mukai to understand changes in biological membranes as affected by
and Arihara 1994). As a result, adhesion, surface hydrophobicity, variation in a medium added with prebiotics (Oldenhof and others
auto-aggregation, and co-aggregation are phenotypic traits that 2005).
provide potential microbial colonization advantages within the Earlier studies on the beneficial effects of synbiotics are based
intestinal tract. on clinical trials (Olah and others 2002; Bengmark 2005) and
There are several physicochemical properties that affect mem- have not examined in-depth information behind the synbiotic ef-
brane characteristics, which help the adherence of Lactobacillus 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-
JFDS-2016-1677 Submitted 10/12/2016, Accepted 12/9/2016. All authors tobacillus are affected by different prebiotics. FT-IR analysis was
are with Food and Nutritional Science, School of Biological Sciences, The Univ. of used to examine the membrane behavior of bacterial cells and
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: changes in the vibrational pattern of surface functional groups in
[email protected]). 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 under strict aseptic condition. Passaging and seeding of Caco-2
separated and the final absorbance was recorded (ODF -600). The cells were done at 80% confluence with proper monitoring of
comparison of initial (ODI -600) with the final absorbance (ODF - their growth.
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:
Assay of adhesion to intestinal epithelial-like cell line
Caco-2 cells were seeded (1 ×104 cell/mL) in 96-well tissue
%Hydrophobicity = [(ODinitial −ODfinal ) /ODinitial ] ×100 culture plates (NUNC, Thermofisher Scientific, Waltham, Mass.,
U.S.A.). Plates were incubated under 95% RH at 37 °C with 5%
Adhesion Assay CO2 supply to attain 80% confluence prior to the adhesion assay.
Overnight bacterial cells grown in mMRS were harvested, washed
Cell culture twice with PBS (pH 7.2), and resuspended in EMEM medium
The adhesive property of Lactobacillus in different prebiotics was containing 10% of FBS without any antibiotic. The initial con-
assayed using colonocyte-like cell lines, Caco-2 (American Type centration of each combination was maintained at approximately
Culture Collection, Manassas, Va., U.S.A.). The culture of cell line 108 CFU/mL. In the meantime, Caco-2 monolayers were also
was carried out as mentioned previously (Gandhi and Shah 2016) washed twice with PBS to eliminate the traces of antibiotics and
with the following modification. Caco-2 cells were cultured in incubated with live bacteria separately and EMEM solution (10:1;
minimum essential medium eagle (EMEM; Sigma-Aldrich) sup- bacterium: eukaryotic cells). The plates were incubated for 120
plemented with 10% fetal bovine serum (FBS) and 1% penicillin- min at 37 °C and 5% CO2 . After incubation, the supernatant
streptomycin solution (Gibco BRL, Grand Island, N.Y., U.S.A.). was discarded and the wells were gently washed with PBS. The at-
The culture was maintained through incubation at 37 °C with tached bacteria were removed from the monolayers by trypsinizing
Food Microbiology &

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).

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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 respectively. Similarly, the greatest degree of adhesion for L. plan-
observed in the case of L. plantarum WCFS-1. This organism tarum WCFS-1 was observed in the medium containing cellobiose
grown in the medium with cellobiose exhibited relatively higher as well (16.85 ± 1.20 %). However, the performance of adhesion
percentage of hydrophobicity (81.93 ± 0.79%) when compared in the other 3 prebiotics was different, with FOS and raffinose
with others. While in the raffinose, it showed the lowest values as ranked at 2nd place (10.73% and 10.97%, respectively) and inulin
well. The result is in accordance with the earlier work by Li and had the lowest values.
others (2010), the carbohydrate source (sucrose) in the growth For L. paracasei 276, the presence of cellobiose not only showed
medium affected the surface parameters henceforth influencing the highest level of adhesion but also contributed a high degree of
membrane hydrophobicity of the bacterium, L. casei Zhang. auto-aggregation and relatively high membrane hydrophobicity
(Table 1; Figure 1). On the other hand, L. paracasei 276 showed the
Adhesion to Caco-2 cells highest degree of hydrophobicity in FOS but exhibited the lowest
Adhesion is considered as an important characteristic for pro- level of attachment. This discrepancy could be due to other factors
biotic bacteria in order to establish colonization in the intestine. such as membrane electrostatic interaction, and steric forces,
In this study, the adhesion ability of Lactobacillus was examined which may have influenced the hydrophobicity of probiotics. For
to understand the influence of different prebiotics on adhesion to L. plantarum WCFS-1, the highest level of attachment in cellobiose
Caco-2 cell (Figure 2). For L. paracasei 276, the highest adhesion also exhibited higher hydrophobicity and auto-aggregation. Sev-
was observed in cellobiose (23.31 ± 0.64 %) followed by inulin eral studies have shown a relationship between aggregation ability,
(13.59 ± 0.53 %). However, the adhesion in the medium with membrane hydrophobicity, and adhesion property (Collado
FOS or raffinose showed the lowest values at 5.9% and 6.18%, 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.
Food Microbiology &
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Potential of probiotics and prebiotics . . .

correlation coefficients between bacterial auto-aggregation Table 4–Infrared band absorption ratio of amide (1400 to 1700
abilities, hydrophobicity, and cell adhesion property are shown in cm−1 ) to sugar (800 to 1200 cm−1 ) of L. paracasei 276 and L.
plantarum WCFS-1.
Table 2. In regards to correlation coefficient for L. paracasei 276,
membrane hydrophobicity exhibited a weak correlation with Amide region area / sugar region area
auto-aggregation and Caco-2 cell adhesion, whereas there was a L. paracasei 276 L. plantarum WCFS-1
very strong correlation between auto-aggregation and Caco-2 ad-
hesion. In L. plantarum WCFS-1, highest degree of correlation was FOS 0.062 0.047
Inulin 0.084 0.054
observed for aggregation and hydrophobicity, meanwhile adhesion Raffinose 0.086 0.113
ability showed medium ranged correlation with auto-aggregation, Cellobiose 0.161 0.110
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 and inulin formed another group (Figure 4B). The spectra ob-
(2014). The authors found the Lactobacillus plantarum exhibiting served from FT-IR are indicative of the changes that occurred in
higher adhesion to epithelial wall had lowest hydrophobicity com- the surfaces of both Lactobacilli.
pared to other Lactobacillus strains. In several studies, hydrophobic- Absorption bands at 2960 cm−1 , corresponding to asymmetric
ity has been shown to be correlated with adhesion ability (Del Re stretching of -CH3 in fatty acids, had no shift in both Lactobacillus
and others 2000), whereas in our study, adhesion ability of both (Table 3). The band between 1700 and 1470 cm−1 referred to
Lactobacillus strains was not in correlation with hydrophobicity of the amide regions of membrane protein structure. The C = O
membrane. As adhesion ability is a more complex assay involving stretching of amide is referred as amide I and observed at 1600 to
several other parameters (surface exo-polysaccharides, S-layer 1700 cm−1 (Santivarangkna and others 2007) while N-H bending
protein, lipoteichoic acid) and hydrophobicity can also contribute at approximately 1554 cm−1 is referred as amide II. In all spectra,
to adhesion ability to some extent. Interestingly, auto-aggregation shifts around 1450 to 1450 cm−1 were observed; these are the
for both strains of Lactobacillus was positively correlated with the characteristic of the scissoring movement of –CH2 groups. Major
adhesion ability, which was similar to that reported in the earlier significant shifts were observed at amide II region of L. paracasei 276
reports (Del and others 2000; Wang and others 2010). (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
FT-IR spectra analysis of Lactobacillus to 800 cm−1 were constantly attributed by the peptidoglycan of
Infrared spectra were recorded for the bacterial cells membrane bacteria. Thus alteration in these area spectra may indicate changes
grown in the medium containing varying prebiotics (Table 3). in the surface structure. Changes in membrane caused by different
The shifts of wavenumbers that occurred at Amide I and Amide growth media can also be explained by comparing the areas under
II regions are shown in Figure 3. The dendogram obtained using specific infrared spectral regions. For L. paracasei 276, cellobiose
Pearson coefficient as the method of generation is presented in had higher amide area/ sugar area ratio, indicating the amount

Food Microbiology &

Figure 4. The data from infrared spectrum clearly show that there of detectable protein was more than the bacterial surface sugar
was a variation in vibration or stretching of bacterial membrane (Table 4). In case of L. plantarum WCFS-1, raffinose and cellobiose

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functional groups. For L. paracasei 276, there was a close relation- had higher ratio of amide area to sugar area (0.113 and 0.11,
ship between the spectral bands when the bacterium was grown respectively). Proteins on the surface of bacteria are an important
in FOS, raffinose, and inulin, whereas the bands in cellobiose was parameter for determining their adhesive nature to the epithelial
distinctly separated from others (Figure 4A). However, in the case wall. As reported by Wang and others (2010), higher amount of
of L. plantarum WCFS-1, spectral bands in raffinose and cellobiose surface protein contributes to the membrane hydrophobicity. As
synbiotic were clustered into one group, while the bands in FOS 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.

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inactivation of surface protein by trypsin (Coconnier and others Gibson GR, Roberfroid MB. 1995. Dietary modulation of the human colon microbiota: intro-
ducing the concept of prebiotics. J Nutri 125:1401–12.
1992). Gourbeyre P, Denery S, Bodinier M. 2011. Probiotics, prebiotics, and synbiotics: impact on the
gut immune system and allergic reactions. J Leukocyte Biol 89(5):685–95.
Conclusion Kankaanpaa PE, Salminen SJ, Isolauri E, Lee YK. 2001. The influence of polyunsaturated fatty
acids on probiotic growth and adhesion. FEMS Microbiol Lett 194:149–53.
The current work found that different prebiotics had varying Kos B, Šušković J, Vuković S, Šimpraga M, Frece J, Matošić S. 2003. Adhesion and aggregation
ability of probiotic strain Lactobacillus acidophilus M92. J Appl Microbiol 94(6):981–7.
effect on surface characteristics like membrane hydrophobicity, Kotzamanidis C, Kourelis A, Litopoulou-Tzanetaki E, Tzanetakis N, Yiangou M. 2010. Evalu-
auto-aggregation, and Caco-2 cell adhesion ability. A further as- ation of adhesion capacity, cell surface traits and immunomodulatory activity of presumptive
sociation was established by Pearson’s correlation coefficient which probiotic Lactobacillus strains. Int J Food Microbiol 140(2):154–63.
Li H, Lu M, Guo H, Li W, Zhang H. 2010. Protective effect of sucrose on the membrane
was supported by the FT-IR data. Both Lactobacillus strains showed properties of Lactobacillus casei Zhang subjected to freeze-drying. J Food Prot 73(4):715–9.
greater adhesion ability in the presence of cellobiose, and in- Martins M, Faleiro ML, da Costa AMR, Chaves S, Tenreiro R, Matos AP, Costa MC. 2010.
Mechanism of uranium (VI) removal by two anaerobic bacterial communities. J Hazard Mater
teresting results from FT-IR also exhibited changes associated 184(1):89–96.
with surface characteristics of Lactobacillus in the presence of this Mukai T, Arihara K. 1994. Presence of intestinal lectin-binding glycoproteins on the cell surface
of Lactobacillus acidophilus. Biosci Biotech Bioch 58(10):1851–4.
prebiotic. Nauman D, Barnickel G, Bradaczek H, Labischinski H, Giesbrecht P. 1982. Infrared spec-
troscopy, a tool for probing bacterial peptidoglycan. Eur J Biochem 125(3):505–15.
Olah A, Belagyi T, Issekutz A, Gamal ME, Bengmark S. 2002. Randomized clinical trial of
References specific Lactobacillus and fibre supplement to early enteral nutrition in patients with acute
Aachary AA, Prapulla SG. 2011. Xylooligosaccharides (XOS) as an emerging prebiotic: microbial pancreatitis. Brit J Surg 89(9):1103–7.
synthesis, utilization, structural characterization, bioactive properties, and applications. Compr Oldenhof H, Wolkers WF, Fonseca F, Passot S, Marin M. 2005. Effect of sucrose and mal-
Rev Food Sci F 10(1):2–16. todextrin on the physical properties and survival of air-dried Lactobacillus bulgaricus: an in situ
Bengmark S. 2005. Synbiotics and the mucosal barrier in critically ill patients. Curr Opin Fourier transform infrared spectroscopy study. Biotechnol Prog 21(3):885–92.
Gastroen 21(6):712–6. Oust A, Møretrø T, Kirschner C, Narvhus JA, Kohler A. 2004. Evaluation of the robustness
Coconnier MH, Klaenhammer TR, Kerneis S, Bernet MF, Servin AL. 1992. Protein-mediated of FT-IR spectra of Lactobacilli towards changes in the bacterial growth conditions. FEMS
adhesion of Lactobacillus acidophilus BG2FO4 on human enterocyte and mucus-secreting cell Microbiol Lett 239(1):111–6.
lines in culture. Appl Environ Microbiol 58(6):2034–9. Sadiq MB, Hanpit hakpong W, Tarning J, Anal AK. 2015. Screening of phytochemicals and in
Collado MC, Meriluoto J, Salminen S. 2008. Adhesion and aggregation properties of probiotic vitro evaluation of antibacterial and antioxidant activities of leaves, pods and bark extracts of
and pathogen strains. Eur food Res Technol 226(5):1065–73. Acacia nilotica (L.) Del. Ind Crop Prod 77:873–82.
Deepika G, Green RJ, Frazier RA, Charalampopoulos D. 2009. Effect of growth time on the Santivarangkna C, Wenning M, Foerst P, Kulozik U. 2007. Damage of cell envelope of Lacto-
surface and adhesion properties of Lactobacillus rhamnosus GG. J Appl Microbiol 107(4):1230– bacillus helveticus during vacuum drying. J Appl Microbial 102(3):748–56.
40. Schachtsiek M, Hammes WP, Hertel C. 2004. Characterization of Lactobacillus coryniformis DSM
Del Re B, Sgorbati B, Miglioli M, Palenzona D. 2000. Adhesion, autoaggregation and hy- 20001T surface protein Cpf mediating coaggregation with and aggregation among pathogens.
drophobicity of 13 strains of Bifidobacterium longum. Lett Appl Microbiol 31(6):438–42. Appl Environ Microbiol 70(12):7078–85.
FAO/WHO. 2002. Guidelines for the Evaluation of Probiotics in Food. Food and Agriculture Schar-Zammaretti P, Dillmann ML, D’Amico N, Affolter M, Ubbink J. 2005. Influence of
Organization of the United Nations and World Health Organization Working Group Report, fermentation medium composition on physicochemical surface properties of Lactobacillus aci-
London Ontario, Canada. dophilus. Appl Environ Microbiol 71:8165–73.
Gandhi A, Shah NP. 2014. Effects of salt concentration and pH on structural and functional Vinderola CG, Medici M, Perdigón G. 2004. Relationship between interaction sites in the
properties of Lactobacillus acidophilus: FT-IR spectroscopic analysis. Int J Food Microbiol gut hydrophobicity, mucosal immunomodulating capacities and cell wall protein profiles in
173:41–7. indigenous and exogenous bacteria. J Appl Microbial 96:230–43.
Gandhi A, Shah NP. 2016. Effect of salt stress on morphology and membrane composition Wang LQ, Meng XC, Zhang BR, Wang, Y, Shang YL. 2010. Influence of cell surface properties
of Lactobacillus acidophilus, Lactobacillus casei, and Bifidobacterium bifidum, and their adhesion to on adhesion ability of bifidobacteria. World J Microb Biot 26(11):1999–2007.
human intestinal epithelial-like Caco-2 cells. J Dairy Sci 99(4):2594–605.
Garcı́a-Cayuela T, Korany AM, Bustos I, de Cadiñanos LPG, Requena T, Peláez C, Martı́nez-
Cuesta MC. 2014. Adhesion abilities of dairy Lactobacillus plantarum strains showing an aggre-
gation phenotype. Food Res Int 57:44–50.
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