Hindawi Publishing Corporation

Journal of Biomedicine and Biotechnology

Volume 2012, Article ID 286216, 6 pages
doi:10.1155/2012/286216

Research Article
Effects of Saponins against Clinical E. coli Strains and
Eukaryotic Cell Line

Michał Arabski,1 Aneta Wȩgierek-Ciuk,2 Grzegorz Czerwonka,1

Anna Lankoff,2, 3 and Wiesław Kaca1
1 Department of Microbiology, Institute of Biology, Jan Kochanowski University in Kielce, ul. Świȩtokrzyska 15, 25-406 Kielce, Poland
2 Department of Radiobiology and Immunology, Jan Kochanowski University in Kielce, ul. Świȩtokrzyska 15, 25-406 Kielce, Poland
3 Center for Biological Densitometry and Radiobiology, Institute of Chemistry and Nuclear Technology, ul. Dorodna 16,

03-195 Warsaw, Poland

Correspondence should be addressed to Michał Arabski, [email protected]

Received 12 September 2011; Accepted 22 November 2011

Academic Editor: Celina Janion

Copyright © 2012 Michał Arabski et al. This is an open access article distributed under the Creative Commons Attribution License,
which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Saponins are detergent-like substances showing antibacterial as well as anticancer potential. In this study, the effects of saponins
from Quillaja saponaria were analyzed against prokaryotic and eukaryotic cells. Multidrug-resistant clinical E. coli strains were
isolated from human urine. As eukaryotic cells, the CHO-K1 cell lines were applied. Antibacterial effect of ampicillin, streptomycin,
and ciprofloxacin in the presence of saponins was measured by cultivation methods. Properties of saponins against CHO-K1 cells
were measured by the MTT test, hemolysis assay and flow cytometry. Saponin from Quillaja saponaria has a cytotoxic effect at
concentrations higher than 25 μg/mL and in the range of 12–50 μg/mL significantly increases the level of early apoptotic cells.
Saponin at dose of 12 μg/mL enhances the six E. coli strains growth. We postulate that saponins increase the influx of nutrients
from the medium into E. coli cells. Saponins do not have synergetic effects on antibacterial action of tested antibiotics. In contrary,
in the presence of saponins and antibiotics, more CFU/mL E. coli cells were observed. This effect was similar to saponins action
alone towards E. coli cells. In conclusion, saponins was cytotoxic against CHO-K1 cells, whereas against E. coli cells this effect was
not observed.

1. Introduction structure) [1]. The biological properties of saponins depend

on the structure of the aglycone and/or the number of sugar
Saponins contain a steroidal or a triterpenoid aglycone to units involved [5]. The therapeutic potential of saponins
which one or more sugar chains are attached [1]. These against eukaryotic cells is associated with their cell mem-
chemical structures determine their biological properties as brane-permeabilizing properties by complexing with choles-
natural nonionic detergents which have cytotoxic, hemo- terol [6]. Our previous studies with a series of synthetic
lytic, molluscicidal, anti-inflammatory, antifungal, antiyeast, saponins showed their potential as anticancer drugs [7]. Sap-
antibacterial, and antiviral activities [2–4]. Saponins are onins, having surface-active properties, might insert into the
nonionic detergents belonging to the group of glycosides. lipid bilayer, bind to cholesterol, form domains enriched with
They are used in the pharmaceutical industry because some cholesterol-saponin complexes, and finally lyse cells [8]. On
forms are the starting point for the semisynthesis of steroidal the other hand, the absence of cholesterol in the membrane
drugs. Many have pharmacological properties and are used structure does not inhibit pore formation by saponins [9,
in phytotherapy and in the cosmetic industry. Saponins can 10]. In our previous study, we suggested that saponin might
be classified into two groups based on the nature of their disturb the permeability of the bacterial outer membrane.
aglycone skeleton: steroidal (consisting of a C27 spirostane About 90% of the surface of naturally cholesterol-free Gram-
skeleton comprising a six-ring structure) and triterpenoid negative bacteria cell-wall outer membranes are covered by
(consisting of a C30 skeleton comprising a pentacyclic lipopolysaccharide (LPS). We concluded that saponin might
2 Journal of Biomedicine and Biotechnology

interact with the lipid A part of Proteus LPSs and thereby USA) described elsewhere [16]. Briefly, the cells were washed
increase the permeability of bacterial cell wall. Lipid A-sap- two times with cold PBS and then resuspended in a 1 × bind-
onin complexes might promote antibiotic (colistin, ampi- ing buffer at a concentration of 1 × 106 cells/mL. An aliquot
cillin) uptake to inherently resistant bacteria cells [11]. of 125 μL of the cell suspension was incubated with 5 μL of
The potentially antibacterial effect of saponins from clin- Annexin V-FITC and 5 μL of propidium iodide (PI) at room
ical point of view is associated with a comprehensive inves- temperature for 15 min in the dark. The cells were resus-
tigation of its antibacterial effect and minimal cytotoxic pro- pended in 400 μL of 1 × binding buffer. The fluorescence was
perties in eukaryotic cells. We previously showed that colistin determined using a Becton Dickinson FACSscan flow cyto-
or ampicillin in the presence of 15 μg/mL saponins reduced meter. A computer system (CellQuest Pro, Becton Dickin-
the numbers of cells of the laboratory strains P. mirabilis son) was used for data acquisition and analysis. Data for
S1959 and R45 [11]. An important medical problem is the 20000 events were stored. A cell gate containing CHO-K1 was
elimination of pathogenic Gram-positive and Gram-negative established on the basis of forward and side light scatter. Four
bacterial strains. Several types of actions are published, such different populations of cells were detected with the Annexin
as bacteriophage treatment [12], inhibition of bacterial adhe- V-FITC kit: normal cells that are Annexin negative and PI ne-
sion [13], and increasing the permeability of bacterial cell gative and express no fluorescence, early apoptotic cells that
walls [14]. are Annexin positive and PI negative and express green flu-
The aims of the present study are orescence, late apoptotic/necrotic cells that are Annexin pos-
itive and PI positive and express green and orange fluores-
(i) testing the saponin abilities to induce hemolytic and cence, and necrotic cells that are Annexin negative and PI
cytotoxic effects against eukaryotic cells, positive and express orange fluorescence.
(ii) studies of saponin influences on growth of clinical
E. coli strains,
2.5. Hemolysis Assay. Blood was obtained from healthy
(iii) testing of saponin abilities to interfere with antibiotic
young male donors. The red blood cells (RBCs) were washed
action against E. coli cells.
three times and resuspended in sterile PBS to give about 15 ×
106 cells per mL and further processed [17]. The erythrocytes
2. Material and Methods were incubated with saponin in a range of 0.5–50 μg/mL for
1 h at 37◦ C. After centrifugation of the nonhemolysed eryth-
2.1. Chemicals. Saponin from Quillaja saponaria bark was rocytes (1400 rpm, 15 min), the absorbance of the released
obtained from Sigma Chemical Co., St. Louis, MO, USA. The hemoglobin in the wavelength range of 400–600 nm was
main aglycone (sapogenin, 20–35%) moiety is quillaic acid, measured. The percentage of hemolysis was determined by
a triterpene of predominantly 30 carbon atoms of the 12 - comparing the absorbance of hemoglobin at 435, 540, and
oleanane type. The aglycone is bound to various sugars, in- 575 nm released from the RBCs in the presence of saponin.
cluding glucose, glucuronic acid, galactose, xylose, apiose, The positive control (100% hemolysis) was determined by
rhamnose, fucose, and arabinose. the amount of hemoglobin released from 15 × 106 RBCs after
10 min of incubation with water.
2.2. Cell Culture. CHO-K1 cells were cultured at 37◦ C
in a humidified 5% CO2 atmosphere in plastic dishes in
McCoy’s 5A medium supplemented with 10% heat-inacti- 2.6. Bacteria Growth in the Presence of Saponin and Antibi-
vated fetal calf serum, 2 mM L-glutamine, and antibiotics otics. Six strains of E. coli isolated from patients at the
(100 units/mL penicillin and 100 μg/mL streptomycin). Department of Microbiology, Holy Cross Cancer Center in
Kielce, Poland, were included in this study. The stock solu-
tion of saponin (Sigma Chemical Co., St. Louis, MO, USA)
2.3. MTT Test. Saponin at final concentrations of 12 to
was diluted in LB medium or minimal M9 medium described
100 μg/mL was added to 500 μl of the CHO-K1 cell suspen-
elsewhere [18]. The reaction mixtures contained saponin at
sion (1.5 × 106 /mL of complete McCoy’s 5A medium) onto
concentrations of 0.1–12 μg/mL and a bacterial suspension
plates. CHO-K1 cells were treated with saponin for 72 hours
of E. coli strains at 102 cells/mL in each probe in final volu-
at 37◦ C in 5% CO2 . After treatment, the viability of cells was
mes of 300 μl of proper medium. Incubation was carried out
evaluated by the MTT assay. MTT reagent was added to each
for 18 h at 37◦ C. Bacterial viability was expressed in terms
plate, and after 6 h of incubation in Hera Cell, 1 mL of SDS
of colony-forming units (CFU/mL). Additionally, the anti-
(10% in 0.01 N HCl) was added to dissolve the water-insolu-
microbial susceptibility of the clinical E. coli strains to ampi-
ble formazan salt. One hour later, the OD650 nm-OD570 nm
cillin, streptomycin, and ciprofloxacin at 25–400 μg/mL was
difference was measured. Unexposed cells were regarded as
prospectively tested in the presence of saponin at 12 μg/mL
100% viable [15].
in LB medium by the cultivation methods.
2.4. Apoptosis. CHO-K1 cells were treated with saponin at
different concentrations (6, 12, 24, and 50 μg/mL) for 72 2.7. Data Analysis. The data were analyzed using the Sta-
hours. After treatment, the frequencies of early apoptotic, tistica software package (StatSoft, Tulsa, OK, USA). All the
late apoptotic, and necrotic cells were evaluated with the values in this study are expressed as the mean ± SD of three
Annexin V-FITC apoptosis detection Kit I (BD Pharmingen, experiments. If no significant differences between variations
Journal of Biomedicine and Biotechnology 3

Table 1: Percentage of early and late apoptotic and necrotic CHO-K1 cells following treatment with saponin measured by flow cytometry;
mean of three experiments ± SD. IP: propidium iodide.

Apoptosis
Saponin (μg/mL) Normal cells (Annexin−/IP−) Necrosis (Annexin−/IP+)
Early (Annexin+/IP−) Late (Annexin+/IP+)
0 96.08 ± 0.07 0.07 ± 0.06 2.53 ± 0.06 0.69 ± 0.06
12 85.75 ± 0.62 5.99 ± 0.33 6.21 ± 0.19 2.06 ± 0.13
25 79.73 ± 2.52 18.67 ± 2.49 1.89 ± 0.04 0.70 ± 0.04
50 64.59 ± 0.32 30.88 ± 0.10 2.09 ± 0.16 2.52 ± 0.17

100
CHO cells viability (%)

80

60 ∗

40
∗
20 ∗

0
0 12 25 50 100
Saponin (µg/mL)

Figure 1: Percentage of the viability of CHO-K1 cells following treatment with saponin measured by the MTT assay; mean of three
experiments ± SD. ∗ P < 0.005.

1.6
∗
the MTT assay. It was found that saponin at concentrations
1.4 25 and 50 μg/mL caused 70% and 50% cell killing, respec-
tively.
1.2
Absorbance

1
3.1.2. Apoptosis. Table 1 shows the level of apoptosis mea-
0.8 ∗ ∗ sured by flow cytometer with the Annexin V-FITC kit and
∗
0.6 ∗ propidium iodide staining according to [16]. Saponin dose-
∗∗ depended lyses of CHO-K1 cells were observed, as in MTT
0.4
∗∗
test (Figure 1). The number of early and late apoptotic cells
0.2 was detected after incubation with saponin in a concentra-
0 tion range of 12–50 μg/mL. The flow cytometric analyses
3 6 12 25 50 100 of the CHO-K1 cells population showed that saponins at
Saponin (µg/mL) concentrations of 12–50 μg/mL significantly increased the
435 nm percentage of early apoptotic cells up to 30.88%. It was in
540 nm a saponin dose-dependent manner compared with the un-
575 nm treated control. The increase in the number (6.21%) of late
apoptosis was notice at the dose of 12 μg/mL of saponin. The
Figure 2: Hemolysis of human erythrocytes incubated with sap-
onin. Hemolysis was measured at 435, 540, and 570 nm; mean of
necrosis of cells varied and did not excite 2.52% at saponin
three independent experiments ± SD. ∗ P < 0.005. dose 50 μg/mL.

3.1.3. Hemolysis Assay. In addition to CHO-K1 cell line, the

were found as assessed by the Snedecor-Fisher test, the human red blood cells (RBCs) were used as hemolysis mark-
differences were compared by the ANOVA test. er. Figure 2 shows the amount of hemoglobin released from
RBCs that was determined by the absorbance at 435, 540, and
3. Results 575 nm in the presence of saponin at concentrations ranging
from 3 to 100 μg/mL. We observed a statistically significant,
3.1. The Effect of Saponin on Eukaryotic Cells dose-depended increase in hemolysis at concentrations from
25 up to 100 μg/mL. The proportion of pick at 435, 540, and
3.1.1. Drug Resistance. Figure 1 shows the percent viability 575 nm was similar. This may suggest that porfirine rings
of the CHO-K1 cells after 4-day treatment with saponin at and iron molecules are not released from hemoglobin, and
concentrations ranging from 12 to 100 μg/mL measured by observed hemolysis is the effect of RBCs wall destruction.
4 Journal of Biomedicine and Biotechnology

1014
∗
1012
1012

1010 1010

(CFU/mL)
(CFU/mL)

108 108

106 106

104 104

102 102
0 0.1 0.2 0.4 0.75 1.5 3 6 12 0 0.1 0.2 0.4 0.75 1.5 3 6 12
Saponin (µg/mL) Saponin (µg/mL)
E. coli strain 1 E. coli strain 4
E. coli strain 2 E. coli strain 5
E. coli strain 3 E. coli strain 6
(a) (b)

Figure 3: The effect of saponin on the growth of the E. coli strains incubated for 18 h at 37◦ C in LB (a) and M9 medium (b); mean of three
independent experiments ± SD. ∗ P < 0.005.

The saponin in range 3–12 μg/mL does not induce hemolysis. 4. Discussion
This indicates that critical amount of saponin molecules
Medically desired situation is the concentration of saponins
25 μg/mL is required to destabilize cell wall of erythrocytes.
with antibacterial properties that has minimal cytotoxic acti-
vity against eukaryotic cells. Saponins possess detergent-like
properties and might increase the permeability of bacterial
3.2. The Effect of Saponin on Bacterial Cells cell membranes without destroying them [19]. In theory, this
activity might facilitate antibiotic influx through the bacte-
3.2.1. The Effect of Saponin on Bacterial Growth. Figure 3(a)
rial cell wall membrane. In the presented studies, we com-
shows the effect of saponin at concentrations ranging from 1
pared saponins’ activities against prokaryotic and eukaryotic
to 12 μg/mL on the growth of the six clinical E. coli strains in- cells. We used commercial saponin from Quillaja saponaria,
cubated for 18 h at 37◦ C in LB medium. The presence of which is one of the major sources of industrial triterpenoid
saponin does not inhibite E.coli cells growth. It is worth to saponins. Saponins extracts from Quillaja saponaria have
notice that we observed statistically significant increases in been in practical use as foaming agents in beverages or emul-
CFU of the multidrug resistant- (MDR-) E. coli strains in the sifiers in foods [20]. In our investigations, we analyzed the
presence of saponin at a concentration of 6 and 12 μg/mL in activity of saponin from Quillaja saponaria against CHO-K1
LB medium, in contrast to growth of bacterial strains in M9 cell line. The saponin extracts increase the number of early
medium which was influenced by saponin presence (Figure apoptotic cells in a dose-dependent manner. Saponins do not
3(b)). induce significantly necrosis and late apoptosis. Effect of sap-
onin on membrane receptors for apoptosis was shown also
by others [5]. In our studies, we also have shown that saponin
3.2.2. The Effect of Saponin and Antibiotics on Bacterial at concentrations higher than 12 μg/mL has significantly cy-
Growth. In next experiment, dose of 12 μg/mL of saponin in totoxic effect on CHO-K1 cells. Similar effect was observed
LB medium was used for testing antibiotic action against E. with human erythrocytes at dose 25 μg/mL of saponins. In-
coli cells. Figure 4 shows the effect of the presence of saponin terestingly, lower doses do not lyse of RBC at all. That may in-
and ampicillin, streptomycin, or ciprofloxacin at concentra- dicate a defined amount of saponins intercalating to cell wall
tions ranging from 25 to 400 μg/mL on the growth of the membranes of erythrocytes needed to lyse it. To analyze
clinical E. coli strains incubated for 18 h at 37◦ C in LB med- the antibacterial properties of saponin, we use saponin at
ium. All six strains were sensitive to ciprofloxacin and resis- 12 μg/mL taking into consideration its minimal cytotoxic ef-
tant to ampicillin and streptomycin. The addition of saponin fect against eukaryotic cells. Saponin does not inhibites E. coli
caused in strains 3 and 6 that ciprofloxacin was less effective growth in minimal M9 and Luria broth. In contrary, we ob-
(Figure 4 compares open circle and open triangle). Having a served statistically significant increases in CFU/mL of six
similar effect, increases of amount of cells were observed with tested MDR E. coli strains in the presence of saponin at a
ampicillin or streptomycin and saponin mixtures in all concentration of 12 μg/mL. This effect was not observed on
strains tested (black or gray circle and triangle, Figure 4). The bacterial growth in M9 minimal medium. Our study of
presented results indicate that saponin enhances clinical E. analysis of susceptibility of clinical E. coli strains against
coli cells growth in the presence of antibiotics. ampicillin, streptomycin, and ciprofloxacin in presence of
Journal of Biomedicine and Biotechnology 5

1010 1010
E. coli strain 1 E. coli strain 2

108 108

(CFU/mL)
(CFU/mL)

106 106

104 104

102 102
0 25 50 100 200 400 0 25 50 100 200 400
Antibiotic (µg/mL) Antibiotic (µg/mL)
(a) (b)

1010 E. coli strain 3 1010

E. coli strain 4

108 108
(CFU/mL)

(CFU/mL)
106 106

104 104

102 102
0 25 50 100 200 400 0 25 50 100 200 400
Antibiotic (µg/mL) Antibiotic (µg/mL)
(c) (d)

1010 1010
E. coli strain 5 E. coli strain 6

108 108
(CFU/mL)

(CFU/mL)

106 106

104 104

102 102
0 25 50 100 200 400 0 25 50 100 200 400
Antibiotic (µg/mL) Antibiotic (µg/mL)
Ampicillin Ampicillin + 12 µg/mL saponin Ampicillin Ampicillin + 12 µg/mL saponin
Streptomycin Streptomycin + 12 µg/mL saponin Streptomycin Streptomycin + 12 µg/mL saponin
Ciprofloxacin Ciprofloxacin + 12 µg/mL saponin Ciprofloxacin Ciprofloxacin + 12 µg/mL saponin
(e) (f)

Figure 4: The effect of saponin at 12 μg/mL and ampicillin, streptomycin, or ciprofloxacin at concentrations ranging from 25 to 400 μg/mL
on the growth of the E. coli strains incubated for 18 h at 37◦ C in LB medium; mean of three independent experiments ± SD.

saponin at a concentration of 12 μg/mL confirmed that sap- due to differences on cell membranes of two cell types.
onins enhance bacteria growth, even in antibiotic presence. The biological properties of chemical substances potentially
In conclusion, we presented varied potencies of saponins useful in clinical use should be characterized by their defined
against eukaryotic and prokaryotic cells. That is probably activity in correlation with the optimal concentration.
6 Journal of Biomedicine and Biotechnology

Acknowledgments [13] B. Montdargent and D. Letourneur, “Toward new biomateri-

als,” Infection Control and Hospital Epidemiology, vol. 21, no. 6,
The work was supported by a Grant no. N N304 044639. The pp. 404–410, 2000.
authors would like to thank Bonita Durnaś from the Depart- [14] L. Mamelli, S. Petit, J. Chevalier et al., “New antibiotic molecu-
ment of Microbiology, Holy Cross Cancer Center in Kielce, les: bypassing the membrane barrier of gram negative bacteria
Poland, for the collected E. coli strains and Marta Kowalik, increases the activity of peptide deformylase inhibitors,” PLoS
Marianna Jagódka, and Katarzyna Pastuszka for technical ONE, vol. 4, no. 7, article e6443, 2009.
support. [15] I. Majsterek, M. Arabski, A. Czechowska et al., “Imatinib
(STI571) inhibits DNA repair in human leukemia oncogenic
tyrosine kinase-expressing cells,” Zeitschrift fur Naturforschung
References C, vol. 61, no. 11-12, pp. 896–902, 2006.
[16] Z. Darzynkiewicz, Current Protocols in Cell Biology, John Wiley
[1] S. G. Sparg, M. E. Light, and J. Van Staden, “Biological activit- & Sons, 2003.
ies and distribution of plant saponins,” Journal of Ethnophar- [17] M. Arabski, K. Gwoździnski, B. Sudak, and W. Kaca, “Effects
macology, vol. 94, no. 2-3, pp. 219–243, 2004. of Proteus mirabilis lipopolysaccharides with different O-poly-
[2] Y. M. Leung, Y. J. Ou, C. Y. Kwan, and T. T. Loh, “Specific inter- saccharide structures on the plasma membrane of human
action between tetrandrine and Quillaja saponins in promot- erythrocytes,” Zeitschrift fur Naturforschung C, vol. 63, no. 5-6,
ing permeabilization of plasma membrane in human leukemic pp. 460–468, 2008.
HL-60 cells,” Biochimica et Biophysica Acta, vol. 1325, no. 2, pp. [18] J. Sambrook and D. W. Russell, Molecular Cloning: A Labora-
318–328, 1997. tory Manual, Cold Spring Harbor Laboratory Press, New York,
[3] S. Sen, H. P. S. Makkar, S. Muetzel, and K. Becker, “Effect of NY, USA, 3rd edition, 1989.
Quillaja saponaria saponins and Yucca schidigera plant extract [19] M. C. Jacob, M. Favre, and J. C. Bensa, “Membrane cell per-
on growth of Escherichia coli,” Letters in Applied Microbiology, meabilisation with saponin and multiparametric analysis by
vol. 27, no. 1, pp. 35–38, 1998. flow cytometry,” Cytometry, vol. 12, no. 6, pp. 550–558, 1991.
[4] C. Bachran, M. Sutherland, I. Heisler, P. Hebestreit, M. F. [20] D. Pelah, Z. Abramovich, A. Markus, and Z. Wiesman, “The
Melzig, and H. Fuchs, “The saponin-mediated enhanced up- use of commercial saponin from Quillaja saponaria bark as a
take of targeted saporin-based drugs is strongly dependent natural larvicidal agent against Aedes aegypti and Culex pip-
on the saponin structure,” Experimental Biology and Medicine, iens,” Journal of Ethnopharmacology, vol. 81, no. 3, pp. 407–
vol. 231, no. 4, pp. 412–420, 2006. 409, 2002.
[5] M. Chwalek, N. Lalun, H. Bobichon, K. Plé, and L. Vout-
quenne-Nazabadioko, “Structure-activity relationships of
some hederagenin diglycosides: haemolysis, cytotoxicity and
apoptosis induction,” Biochimica et Biophysica Acta, vol. 1760,
no. 9, pp. 1418–1427, 2006.
[6] H. Gögelein and A. Hüby, “Interaction of saponin and
digitonin with black lipid membranes and lipid monolayers,”
Biochimica et Biophysica Acta, vol. 773, no. 1, pp. 32–38, 1984.
[7] H. Myszka, D. Bednarczyk, M. Najder, and W. S. Kaca, “Syn-
thesis and induction of apoptosis in B cell chronic leukemia by
diosgenyl 2-amino-2-deoxy-β-D-glucopyranoside hydrochlo-
ride and its derivatives,” Carbohydrate Research, vol. 338, no.
2, pp. 133–141, 2003.
[8] E. Baumann, G. Stoya, A. Völkner, W. Richter, C. Lemke, and
W. Linss, “Hemolysis of human erythrocytes with saponin
affects the membrane structure,” Acta Histochemica, vol. 102,
no. 1, pp. 21–35, 2000.
[9] R. Segal, P. Shatkovsky, and I. Milo Goldzweig, “On the mech-
anism of saponin hemolysis. I. Hydrolysis of the glycosidic
bond,” Biochemical Pharmacology, vol. 23, no. 5, pp. 973–981,
1974.
[10] W. P. Winter, “Mechanism of saponin induced red cell hemol-
ysis: evidence for the involvement of aquaporin CHIP28,”
Blood, vol. 84, supplement 1–10, abstract 445, 1994.
[11] M. Arabski, S. Wasik, K. Dworecki, and W. Kaca, “Laser inter-
ferometric and cultivation methods for measurement of co-
listin/ampicilin and saponin interactions with smooth and
rough of Proteus mirabilis lipopolysaccharides and cells,” Jour-
nal of Microbiological Methods, vol. 77, no. 2, pp. 178–183,
2009.
[12] S. O’Flaherty, R. P. Ross, and A. Coffey, “Bacteriophage and
their lysins for elimination of infectious bacteria: Review ar-
ticle,” FEMS Microbiology Reviews, vol. 33, no. 4, pp. 801–819,
2009.
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