Biotechnology and Bioprocess Engineering 16: 971-977 (2011)
DOI 10.1007/s12257-011-0050-6
RESEARCH PAPER
Antibacterial Properties of the Extract of Abelmoschus esculentus
Carla C. C. R. de Carvalho, Priscila Almeida Cruz, M. Manuela R. da Fonseca, and Lauro Xavier-Filho
Received: 7 February 2011 / Revised: 29 March 2011 / Accepted: 3 April 2011
© The Korean Society for Biotechnology and Bioengineering and Springer 2011
Abstract In this study, antimicrobial properties of both
lyophilized and fresh water extracts of the okra pods were
assessed against Rhodococcus erythropolis and R. opacus,
Mycobacterium sp. and M. aurum, Staphylococcus aureus,
Escherichia coli, and Xanthobacter Py2. The extracts were
effective against all bacterial strains tested, except R.
erythropolis and the fresh extract displayed better anti-
microbial properties than the lyophilized extract. A fresh
extract concentration of 97.7 mg/mL was sufficient to
kill all S. aureus cells, which is a worldwide source of
nosocomial infection. The extract was also effective in
inhibiting the growth of both Mycobacterium strains, X.
Py2 and S. aureus, but was ineffective against R. eryth-
ropolis and E. coli. The lipid fraction of the okra gum was
found to be responsible for the antibacterial properties and
the protein and polysaccharide fractions displayed no
antimicrobial activity. The two major constituents of the
lipid fraction, palmitic and stearic acids, were apparently
responsible for the antimicrobial properties of the okra
extract.
Keywords: antibacterial agent, biocide, Hibiscus esculentus,
okra, lady finger
Carla C. C. R. de Carvalho, M. Manuela R. da Fonseca
IBB-Institute for Biotechnology and Bioengineering, Centre for Biological
and Chemical Engineering, Instituto Superior Técnico, Lisbon 1049-001,
Portugal
Carla C. C. R. de Carvalho*, M. Manuela R. da Fonseca
Department of Bioengineering, Instituto Superior Técnico, Universidade
Técnica de Lisboa, Lisbon 1049-001, Portugal
Tel: +351-21-841-9189; Fax: +351-21-841-9062
E-mail: ccarvalho@ist.utl.pt
Priscila Almeida Cruz, Lauro Xavier-Filho
Instituto de Tecnologia e Pesquisa, Aracaju-Sergipe 49032-490, Brazil
1. Introduction
The plant Abelmoschus esculentus (L.) Moench, formally
known as Hibiscus esculentus L. (common name okra) is
widely spread from West Africa to India, Southern Europe
and the Americas since it is tolerant to both (i) hot and dry,
and (ii) hot and humid climates [1]. The simplicity of
cultivation techniques and dependable yields make this one
of the best vegetables for the Tropics.
The water extract of A. esculentus fruits, also known as
okra gum, has been efficiently used as a lubricant [2] and
to stabilize foams [3] and suspensions [4]. The gum of the
okra pods has been traditionally used in culinary appli-
cations as a thickener of soups (e.g. Louisiana’s Gumbo
soup) and stews. It has also been used as a substitute of
egg-white and as a replacement of fat in chocolate bar
cookies [5,6] and in chocolate frozen dairy desserts while
maintaining acceptable sensory characteristics of the pro-
ducts [7].
Several previous studies have examined the physical and
chemical properties of the mucilage of okra. In 1954,
Whistler and Conrad found that the polysaccharide solution
obtained from okra consisted of D-galactose, L-rhamnose,
and D-galacturonic acid [8]. By stopping hydrolysis at the
appropriate time, these authors were able to obtain a
crystalline galactobiose, which was shown to be 4-O-D-
galactopyranosyl-D-galactopyranose [9]. In 1956, Amin
identified three carbohydrates and arabinose [10]. In 1973,
BeMiller reviewed previous papers that described the
composition and properties of the mucilage of okra [11].
Four years later, Woolfe and co-workers evaluated the
composition and properties of the okra extract [3]. Hydro-
lysis of the polysaccharide was shown to produce galac-
turonic acid, and the neutral sugars galactose, rhamnose
and glucose at a proportion of 1.3:1.0:0.1:0.1. The proteins
in the extract were not significantly affected by the
purification steps and could not be separated from the
972
polysaccharide by either gel chromatography or disc electro-
phoresis. The moisture content of the crude extract was
9.35% of wet weight, while the protein content was 9.42%.
Mg, Ca, K, and P were present at concentrations of 0.51,
2.02, 0.68, and 0.18% of the wet weight, respectively.
Only a few studies have examined the pharmacological
properties of A. esculentus pods. Ansari et al. [12] evaluated
the antioxidant properties of okra extracts while Lengsfeld
and co-workers [13] studied the ability of the glycosylated
compounds from okra fruits to inhibit the adhesion of
Helicobacter pylori cells to human gastric mucosa. In the
later study, fractioning of the crude polysaccharide extract
showed that the active fractions contained rhamnogalac-
turonans and a considerable amount of glucuronic acid and
also glycosylated proteins with molecular weights between
25 and 37 kDa. Curiously, none of the isolated fractions
was efficient in inhibiting H. pylori adhesion. Based on
these findings, the authors concluded that the anti-adhesive
properties of okra were derived from the combination of
glycoproteins and highly acidic sugar compounds that
make a complex three-dimensional structure only in the
fresh extract of the okra fruits. In Asian medicine, okra
pods have been traditionally used for the treatment of
gastric irritations and inflammation. Other medical appli-
cations include replacement of blood plasma [14]. Bevans
and co-workers previously evaluated the efficacy of the
crashed fruits of okra for the treatment of conjunctivitis and
the fruits and flowers were shown to slightly inhibit
Bacillus cereus and P. aeruginosa growth, while the leaves
were inefficient in inhibiting growth of these bacteria [15].
The aim of the present study was to evaluate the anti-
bacterial properties of the water extract of the okra fruits
against Gram-positive and Gram-negative strains, namely
Rhodococcus and Mycobacterium strains, Staphylococcus
aureus, E. coli, and Xanthobacter Py2.
2. Materials and Methods
2.1. Strains and plant material
Rhodococcus erythropolis DCL14, R. opacus PWD4,
Mycobacterium sp. NRRL B-3805, Mycobacterium aurum
NgR9, Staphylococcus aureus ATCC 25923, Escherichia
coli BL21 (DE3)pLys, Xanthobacter Py2, and Pseudomonas
aeruginosa 8821 were used in this study. The okra pods
were bought in the public market of the city of Aracaju,
Sergipe state, Brazil.
2.2. Bacterial growth
The rhodococci strains and X. Py2 were grown in orbital
incubators at 30ºC, while the remaining strains were grown
at 37º. The agitation speed used in all cases was 200 rpm.
Biotechnology and Bioprocess Engineering 16: 971-977 (2011)
The cells were grown in elemental mineral medium [16]
with phosphate buffer (pH 7.0) and 2 g/L of glucose as the
carbon source except when mentioned otherwise.
2.3. Characterisation of the okra extract
2.3.1. Preparation
After being washed, the okra pods were cut in transversal
half-centimeter slices and immersed in distilled water. The
polysaccharide extract was separated from the remaining
crushed material by sieving and stored at −20ºC. Extrac-
tions were performed with both unpeeled and peeled okra
pods so that a chloroplast-containing and a chloroplast-free
polysaccharide extract were obtained.
2.3.2. Fractioning
A crude protein extract was prepared from okra pods by
ammonium sulphate precipitation and resuspension in
water. The protein content was determined using the Brad-
ford method [17].
The lipids and polysaccharides were extracted after
okra pods were first heated at 70ºC for 10 min to inactivate
the proteins. Extraction with ethanol, which is a polar
solvent, allowed for the extraction of polar lipids present
in the extract and simultaneously the precipitation of the
polysaccharides. Filtration was used to recover the poly-
saccharides from the ethanol phase. Extraction with
chloroform resulted in the extraction of apolar lipids.
After evaporation of the respective solvents under a
stream of nitrogen, both the polar and apolar lipids were
recovered.
2.3.3. Fatty acid composition
The lipids of both the fresh and lyophilised okra extract
were extracted according to the Bligh and Dyer method as
reported by Findlay et al. [18]. The fatty acids were
derivatized into fatty acid methyl esters and analyzed by
gas chromatography on an Agilent (Santa Clara, CA, USA)
6890N gas chromatograph with a FID and a 7683 B series
injector equipped with a 30 m HP-5 capillary column from
J&W Scientific (Folsom, CA, USA), as described previ-
ously [19]. Peak identification was achieved using a
qualitative standard of bacterial fatty acid methyl esters
and one of polyunsaturated fatty acids, both from Supelco
(Bellefonte, USA), and a mixture of cis/trans 18:1 from
Sigma-Aldrich (Buchs, Switzerland). The results presented
in this work were the average of three independent samples.
2.3.4. Surface tension
The surface tension was measured using a plate device in
a K8 tensiometer from Krüss GmbH (Hamburg, Germany)
according to the Wilhelmy technique [20].
Antibacterial Properties of the Extract of Abelmoschus esculentus
2.4. Antibacterial activity of the extract and mechanisms
of action
2.4.1. Effect on bacterial cell viability
Different concentrations of the okra extract were added to
bacterial suspensions of each bacterium. After incubation
for 30 min at 30ºC and 200 rpm, cellular viability was
assessed by fluorescent microscopy and image analysis.
Cell viability was measured using a LIVE/DEAD® Bac-
LightTM bacterial viability kit, which was purchased from
Molecular Probes (Invitrogen Co., Carlsbad, California, USA).
The samples were prepared according to the protocol
provided by the manufacturer and observed by fluore-
scence microscopy using an Olympus CX40 microscope,
with an Olympus U-RFL-T burner and an U-MWB mirror
cube unit (excitation filter: BP450-480; barrier filter:
BA515). Images were captured by an EvolutionTM MP5.1
CCD colour camera using the software Image-Pro Plus,
both of which were purchased from Media Cybernetics,
Inc. Image analysis was carried out as described previously
[21]. At least 15 images of each sample were taken.
2.4.2. Effect on bacterial growth
The ability of the extract to inhibit bacterial growth was
determined by diffusion of the compounds on agar plates
and by broth microdilution susceptibility tests according to
the CLSI guidelines [22]. The diffusion test was performed
by measuring the inhibition area around a well containing
only the extract (100 µL at 100%) on agar plates contain-
ing 104 ~ 105 cell of each strain. To assess which fraction
was responsible for the antibacterial properties, fractions
containing proteins, polysaccharides, polar lipids and apolar
lipids were added to different wells on the agar plate. The
concentration of each fraction used was equal to the con-
centration of each component in the original extract. The
inhibition area was determined by analyzing images using
the software Image-Pro Plus from Media Cybernetics, Inc.
(USA). The area presented did not include the area of the
well. The minimum inhibition concentration (MIC) was
determined by the broth microdilution method in 96-well
microtitre plates. Briefly, twofold serial dilutions of the
okra extract were prepared in Mueller-Hinton broth to a
final volume of 150 µL. The bacterial strains were grown
overnight, collected in the exponential phase and diluted in
PBS to a McFarland 0.5 turbidity standard and 50 µL of
the cell suspension was added to each well. Growth was
assessed after 16 and 18 h. The MIC was defined as the
lowest compound concentration that resulted in no growth.
2.4.3. DNA denaturation
Crude DNA was extracted from bacteria by alkaline lysis
[23]. The A. esculentus extract was mixed in equal pro-
973
portion with the crude DNA and incubated for 45 min.
DNA denaturation was determined after 15 and 45 min of
incubation by UV spectrophotometry and by visually assess-
ing the existence of DNA precipitation.
2.4.4. Protein precipitation
Bacterial proteins were separated from cell lysates by am-
monium sulphate precipitation and resuspended in water.
Protein precipitation was assessed after mixing the protein
extract with the okra extract for 15 min.
2.4.5. Enzyme denaturation
The S. aureus and R. erythropolis cells were collected at
the exponential phase at an optical density of 0.8 and
enzymes were collected by centrifugation after alkaline
lysis of the cells. Catalase activity was determined in the
presence and absence of the A. esculentus extract by mea-
suring the disappearance of hydrogen peroxide at 240 nm
at 30 sec intervals [24].
2.5. Error analysis
The average error associated with the GC quantification of
each FAME was ± 5.1%, with a confidence interval of
99.5%. The errors were calculated based on seven indepen-
dently prepared standard solutions. The error associated
with image analysis was ± 5.8%, based on the standard
deviation and sample mean of fifteen repeated images
taken from the same sample, with a confidence interval of
99.5%.
3. Results and Discussion
In this study, the effects of the okra gum on both cell
viability and bacterial growth were evaluated. Previous
studies have shown that the glycosylated compounds of the
immature fruits are able to inhibit the adhesion of the
bacterium H. pylori to human gastric mucosa in situ [13]
and the adhesion of Campylobacter jejuni to intestinal
epithelia from chicken in vitro [25].
3.1. Properties and composition of the A. esculentus
extract
The density of the extract of the Brazilian A. esculentus
used in this work was 1.024 ± 0.003 g/mL and the pH of
the concentrated crude extract was 4.2 for the peeled okra
and 6.2 for the unpeeled okra. Saturation of lyophilized
extract in water was achieved at a concentration of 9.5 mg/
mL. Solutions containing 0.5, 1, and 10% (v/v) extract had
surface tensions of 69.2, 60.7, and 41.4 mN/m, respec-
tively, indicating that the extract can act as a biosurfactant.
The extract obtained from peeled and unpeeled okra con-
974 Biotechnology and Bioprocess Engineering 16: 971-977 (2011)

Table 1. Minimum inhibitory concentration of okra extract against

several Gram-positive and Gram-negative bacteria
MIC (%, v/v)
Peeled Unpeeled
S. aureus 6.25 6.25
Mycobacterium sp. 12.5 12.5
M. aurum 6.25 6.25
R. erythropolis 66.7 80
R. opacus 50 50
X. Py2 12.5 12.5
E. coli 80 80
P. aeruginosa 12.5 6.25

Fig. 1. Fatty acid composition of the unpeeled, peeled, and

lyophilised okra.
tained, per gram, 5.20 ± 0.01 and 4.89 ± 0.02 g of protein.
The okra extract contained 34.1% palmitic acid and 26.2%
stearic acid, as the main fatty acids, and only 3.8% of
linoleic acid. Similarly, the extract from peeled okra fruits
contained 45.6% palmitic acid, 29.9% stearic acid, and
3.4% of linoleic acid (Fig. 1). However, the lyophilized
extract contained 31.1% of palmitic acid and 21.5% of
linoleic acid since the extract was obtained by crushing the
okra pods with seeds. The fatty acid composition has also
been shown to vary during plant growth [26].
Thin layer chromatography (TLC) indicated that the
polysaccharide contained galactose, rhamnose and glucose.
The composition of the okra gum has been studied in-depth
by several research groups and the major cell wall poly-
saccharides from okra pods were shown to be rich in galac-
tose, rhamnose and galacturonic acid [27]. In addition,
pectin was shown to contain an unusual substitution of its
rhamnosyl residues with acetyl and alpha-linked galactosyl
groups [28]. The major antioxidants in okra were determin-
ed to be quercetin 3-O-xylosyl (1''' → 2'') glucoside,
quercetin 3-Oglucosyl (1''' → 6'') glucoside, quercetin 3-O-
glucoside and quercetin 3-O-(6''-O-malonyl)-glucoside [29].
The amount of magnesium, calcium, potassium and phos-
phorous (0.51, 2.02, 0.68, and 0.18% of wet weight,
respectively) has also been determined [3].
Tamarindus indica had MICs of 8, 15.5, and 14 mg/mL
against S. aureus, E. coli, and P. aeruginosa, respectively
[31]. The fruit pulp of tamarind has also been traditionally
used to make jams, chutneys, desserts and drinks. The
ethanolic extract of leaves and fruits of Hyaenanche globosa
had a MIC of 3.1 mg/mL against M. Smegmatis; however,
this South African plant is poisonous [32].
Cell growth inhibition was also assessed when the cells
were grown on agar plates containing wells filled only with
the okra extract. Cells that were inhibited could not grow
in the area surrounding the well through which the extract
diffused. M. aurum, X. Py2, and P. aeruginosa cells were
largely inhibited and were unable to grow in a circular area
of 343, 306, and 357 mm2, respectively (Fig. 2). The ex-
tract was also efficient in inhibiting the growth of S. aureus
and Mycobacterium sp. However, the growth of R. erythro-
polis and E. coli cells was unaffected and R. opacus cells
were only slightly affected (Fig. 2).
Practically no differences in antibacterial properties were
observed between the okra gum extracted from peeled and

3.2. Antibacterial properties of the A. esculentus extract

Low extract concentrations for both peeled and unpeeled
A. esculentus were effective against S. aureus, Mycobac-
terium sp., M. aurum, X. Py2 and P. aeruginosa (Table 1).
The MICs in these cases ranged between 6.25 and 12.5%
(v/v), which corresponded to 6.4 ~ 12.8 mg/mL. Similar
MIC values have been observed for other plant extracts.
Acetone, methanol, ethanol and both cold and hot aqueous Fig. 2. Inhibition areas observed around wells containing only
extracts of the fruits of Terminalia chebula also had a MIC (100%) peeled and unpeeled A. esculentus extract in agar plates
of 12.5 mg/mL against S. aureus [30]. Ethanolic extracts of with each of the test strains.
Antibacterial Properties of the Extract of Abelmoschus esculentus 975

Fig. 3. Viability of bacterial cells incubated with (A) lyophilized and (B) fresh okra extracts for 30 min.

unpeeled fruits (Table 1 and Fig. 2). Therefore, the remain- Table 2. Viability of R. erythropolis DCL14 and R. opacus PWD4
ing tests were conducted with water extracts obtained from cells grown on glucose, limonene, and toluene as the sole carbon
sources
unpeeled fruits since they are usually used in culinary
recipes. Cell viability (%)
The effect of the okra gum was also evaluated after Okra extract, % (v/v)
exposing resting cells to the extract for 30 min. The gum Carbon source 0.5 1 10
displayed antibacterial properties against all bacterial strains Glucose 100.00 98.10 96.51
tested, as shown by the decrease in cell viability, except R. erythropolis Limonene 97.42 86.03 59.17
towards R. erythropolis cells (Fig. 3). At the same con- Toluene 94.21 75.31 66.10
centration, the fresh extract was, in general, more effective Glucose 98.00 95.21 93.31
than the lyophilized okra extract. In addition, because of R. opacus Limonene 91.37 85.38 53.83
the low solubility of the later, concentrations higher than Toluene 92.42 79.22 37.46
9.5 mg/mL could not be tested. When the results obtained
at fresh extract concentrations of 4.9 and 97.7 mg/mL were
compared, the reduction in cell viability was shown to (v/v), DCL14 and PWD4 cells grown in limonene as the
range from 34.7 to 46.8%. However, exceptions were sole carbon source were also similarly resistant to the okra
observed for R. erythropolis, which was only reduced by a extract, with 59.2 and 53.8% of the cells remaining viable,
3.5%. In addition, for S. aureus and P. aeruginosa, an okra respectively. However, R. opacus PWD4 cells were more
extract concentration of 97.7 mg/mL was completely effec- susceptible than those of DCL14 when grown on toluene.
tive in inhibiting bacterial growth. S. aureus can cause We believe these results occurred because (i) the properties
several illnesses from skin infections to pneumonia, men- and composition of the cellular membrane of the bacterial
ingitis and septicaemia and is the major cause of noso- strains is highly dependent on the carbon source used
comial infections worldwide [33], while P. aeruginosa is [34,35], and (ii) the effect of okra gum was influenced by
known for causing fatal lung infections in patients with the carbon source. These combined results indicate that the
cystic fibrosis. main effects of the extract were on the cellular membrane
As shown in Fig. 3, a significant difference in cell and cell wall.
viability between the two rhodococci strains was observed.
Since R. erythropolis was grown in glucose and R. opacus 3.3. Effect on bacterial macromolecules
was grown in toluene (the latter grows faster on toluene The extract of A. esculentus did not cause visual precipi-
than on glucose), the effect of the carbon source on cell tation of bacterial proteins. The addition of the extract to
susceptibility to the okra extract was evaluated. When the S. aureus and R. erythropolis lysates resulted in only a
cells were grown on glucose, limonene and toluene, the 14.9% reduction of catalase activity indicating that no
viability of R. erythropolis and R. opacus cells, after expo- significant enzymatic inactivation was caused by the
sure to the okra gum, was dependent on the carbon source extract. Similarly, the increase in UV absorbance of crude
used for cell growth (Table 2). Both R. erythropolis DCL14 DNA was only 15.1% after 45 min of incubation with the
and R. opacus PWD4 cells grown on glucose were nearly extract, which suggests that the extract does not cause
unaffected by the okra extract. At a concentration of 10% significant denaturation of crude DNA.
976
Fig. 4. Inhibition areas observed around wells containing only
okra extract, polar and apolar lipids extracted from A. esculentus at
the same concentration used in the okra extract, and olive oil, on
agar plates with S. aureus, M. aurum, R. erythropolis, and E. coli
growing cells. No column indicates that no inhibition was
observed.
3.4. Effect of the fractions of the extract on antibacterial
activity
To assess which components (polysaccharides, proteins or
lipids) were responsible for the antibacterial properties, the
extract was fractioned. Wells made on agar plates were
filled with 100 µL of each fraction containing a concent-
ration equal to that observed in the original extract. The
inhibition areas observed with the original water extract
and that obtained with the polar lipids were very similar;
1.0, 3.4, and 5.7% difference for S. aureus, M. aurum, and
E. coli, respectively (Fig. 4). On the other hand, the inhibi-
tion area when apolar lipids were used was half the value
obtained when the unfractionated extract was used. None
of the extracts was effective against R. erythropolis cells.
Curiously, the growth of this bacterium was affected by
olive oil (Fig. 4), which is mainly composed of oleic (ca.
72%) and linoleic (ca. 21%) acids [36]. However, olive oil
was less effective than the water and ethanol extracts of A.
esculentus against the remaining bacterial strains tested. No
antibacterial effect was observed in the presence of the
fractions containing the proteins and the polysaccharides
alone.
Comparison of the antimicrobial properties of the okra
gum in fractions containing proteins, polysaccharides and
lipids indicate that the components responsible for the
antibacterial properties were lipidic. The short-chain caprylic,
capric, and lauric fatty acids have been found to have
antibacterial properties against mastitis pathogens [37], S.
aureus [38], and Listeria monocytogenes [39]. Long-chain
unsaturated fatty acids have also been shown to display
antibacterial activity probably because they affect fatty acid
synthesis in bacteria. Linoleic, palmitoleic, oleic, linolenic
and arachidonic acids inhibit the enoyl-acyl carrier protein
Biotechnology and Bioprocess Engineering 16: 971-977 (2011)
reductase (FabI), which is essential in bacterial fatty acid
synthesis [40]. The concentration of linoleic acid in the
lyophilized extract in the present study, which was
obtained from okra pods with seeds, was sufficient to
confer antimicrobial properties to the extract.
The major lipids present in the fresh okra extract were
palmitic acid (34.1%) and stearic acid (26.2%) and they
were most likely responsible for the antibacterial properties
of this extract. Palmitic acid was previously found to be the
responsible for the antibacterial properties of Pentanisia
prunelloides, a perennial herb, which root is used in Zulu
traditional medicine [41]. Stearic acid was among the fatty
acids found to confer antibacterial properties to Dombeya
rotundifolia, which has been used in traditional medicine to
treat intestinal ulcers and stomach illnesses [42].
4. Conclusion
The results of this work demonstrate the potential of using
the extract of A. esculentus as an antibacterial agent with
possible applications in the food and pharmaceutical
industries.
Acknowledgements
Carla C. C. R. de Carvalho would like to thank Fundação
para a Ciência e a Tecnologia, Portugal, for financial
support (through the program Ciência 2007).
References
1. Martin, F. W. and R. M. Ruberté (1978) Vegetables for the hot,
humid tropics: Part 2. Okra. U. S. Department of Agriculture,
New Orleans.
2. O'bryant, J. C. (1998) Organic lubricant. US Patent 6,124,248.
3. Woolfe, L. M., M. Chaplin, and G. Otchere (1977) Studies on the
mucilages extracted from Okra fruits and Baobab leaves. J. Sci.
Fd. Agric. 28: 519-529.
4. Wahi, S. P., V. D. Sharma, V. K. Jain, and P. Sinha (1985) Studies
on suspending property of mucilages of Hygrophila spinosa and
Hibiscus esculentus. Indian Drugs 22: 500-502.
5. Costantino, A. J. and J. E. Romanchik-Cerpovicz (2004) Physical
and sensory measures indicate moderate fat replacement in fro-
zen dairy dessert is feasible using okra gum as a milk-fat ingre-
dient substitute. J. Am. Diet. Assoc. 104: 44.
6. Romanchik-Cerpovicz, J. E., R. W. Tilmon, and K. A. Baldree
(2002) Moisture retention and consumer acceptability of choco-
late bar cookies prepared with okra gum as a fat ingredient sub-
stitute. J. Am. Diet. Assoc. 102: 1301-1303.
7. Romanchik-Cerpovicz, J. E., A. C. Costantino, and L. H. Gunn
(2006) Sensory evaluation ratings and melting characteristics
show that okra gum is an acceptable milk-fat ingredient substi-
tute in chocolate frozen dairy dessert. J. Am. Diet. Assoc. 106:
594-597.
Antibacterial Properties of the Extract of Abelmoschus esculentus
8. Whistler, R. L. and H. E. Conrad (1954) 2-O-(D-galactopyrano-
syluronic acid)-L-rhamnose from okra mucilage. J. Am. Chem.
Soc. 76: 3544-3546.
9. Whistler, R. L. and H. E. Conrad (1954) A crystalline galactobi-
ose from acid hydrolysis of okra mucilage. J. Am. Chem. Soc. 76:
1673-1674.
10. Amin, E. S. (1956) The mucilages of Hibiscus esculentus (okra
or bamia-fellahi) and Corchorus olitorius (mulukhia). J. Chem.
Soc. 78: 828-832.
11. BeMiller, J. and R. Whistler (1992) Industrial gums. 3rd ed.,
Academic Press, London, UK.
12. Ansari, N. M., L. Houlihan, B. Hussain, and A. Pieroni (2005)
Antioxidant activity of five vegetables traditionally consumed by
south-Asian migrants in Bradford, Yorkshire, UK. Phytother.
Res. 19: 907-911.
13. Lengsfeld, C., F. Titgemeyer, G. Faller, and A. Hensel (2004)
Glycosylated compounds from okra inhibit adhesion of Helico-
bacter pylori to human gastric mucosa. J. Agric. Food Chem. 52:
1495-1503.
14. Benjamin, B. H., K. H. Ihrig, and D. A. Roth (1951) The use of
okra as a plasma replacement. Rev. Canad. Biol. 10: 215-221.
15. Bevans, N., M. Alford, F. Guanchez, M. Aregullin, and R. Rod-
riguez (2001) Preliminary phytochemical study of two Caribbean
Malvaceae used in the treatment of conjunctivitis. J. Undergrad.
Study Indep. Res. Winter 1: 20-24.
16. Wiegant, W. M. and J. A. M. de Bont (1980) A new route for eth-
ylene glycol metabolism in Mycobacterium E44. J. Gen. Micro-
biol. 120: 325-331.
17. Bradford, M. M. (1976) A rapid and sensitive method for the
quantification of microgram quantities of protein utilizing the
principle of protein dye binding. Anal. Biochem. 72: 248-254.
18. Findlay, R. H., G. M. King, and L. Watling (1989) Efficacy of
phospholipid analysis in determining microbial biomass in sedi-
ments. Appl. Environ. Microbiol. 55: 2888-2893.
19. de Carvalho, C., V. Fatal, S. Alves, and M. da Fonseca (2007)
Adaptation of Rhodococcus erythropolis cells to high concentra-
tions of toluene. Appl. Microbiol. Biotechnol. 76: 1423-1430.
20. Adamson, A. W. and A. P. Gast (1997) Physical chemistry of sur-
faces. 6th ed., Wiley-Interscience, NY.
21. de Carvalho, C. C. C. R., M. -N. E. Pons, M. Manuela, and R. D.
Fonseca (2003) Principal components analysis as a tool to sum-
marise biotransformation data: Influence on cells of solvent type
and phase ratio. Biocatal. Biotransform. 21: 305-314.
22. CLSI (2010) Performance standards for antimicrobial suscepti-
bility testing; Twentieth informational supplement. M100-S20.
23. Birnboim, H. C. and J. Doly (1979) A rapid alkaline extraction
procedure for screening recombinant plasmid DNA. Nucleic
Acids Res. 7: 1513-1523.
24. Beers, R. F. and I. W. Sizer (1952) A spectrophotometric method
for measuring the breakdown of hydrogen peroxide by catalase.
J. Biol. Chem. 195: 133-140.
25. Lengsfeld, C., G. Faller, and A. Hensel (2007) Okra polysaccha-
rides inhibit adhesion of Campylobacter jejuni to mucosa iso-
lated from poultry in vitro but not in vivo. Anim. Feed Sci.
Technol. 135: 113-125.
26. Gopalakrishnan, N., T. N. B. Kaimal, and G. Lakshminarayana
(1982) Fatty acid changes in Hibiscus esculentus tissues during
growth. Phytochem. 21: 565-568.
977
27. Sengkhamparn, N., R. Verhoef, H. A. Schols, T. Sajjaanantakul,
and A. G. J. Voragen (2009) Characterisation of cell wall polysac-
charides from okra (Abelmoschus esculentus (L.) Moench). Car-
bohydr. Res. 344: 1824-1832.
28. Sengkhamparn, N., E. J. Bakx, R. Verhoef, H. A. Schols, T. Saj-
jaanantakul, and A. G. J. Voragen (2009) Okra pectin contains an
unusual substitution of its rhamnosyl residues with acetyl and
alpha-linked galactosyl groups. Carbohydr. Res. 344: 1842-1851.
29. Shui, G. and L. L. Peng (2004) An improved method for the anal-
ysis of major antioxidants of Hibiscus esculentus Linn. J. Chro-
matogr. A. 1048: 17-24.
30. Aneja, K. R. and R. Joshi (2009) Evaluation of antimicrobial
properties of fruit extracts of Terminalia chebula against dental
caries pathogens. Jundishapur J. Microbiol. 2: 105-111.
31. Doughari, J. H. (2006) Antimicrobial activity of Tamarindus
indica Linn. Tropical J. Pharmaceut. Res. 5: 597-603.
32. Momtaz, S., N. Lall, A. Hussein, S. N. Ostad, and M. Abdollahi
(2010) Investigation of the possible biological activities of a poi-
sonous South African plant; Hyaenanche globosa (Euphorbi-
aceae). Pharmacognosy Magazine 6: 34-41.
33. Falcone, M., P. Serra, and M. Venditti (2009) Serious infections
due to methicillin-resistant Staphylococcus aureus: An evolving
challenge for physicians. Eur. J. Intern. Med. 20: 343-347.
34. de Carvalho, C. C. C. R., B. Parreño-Marchante, G. Neumann, M.
M. R. da Fonseca, and H. J. Heipieper (2005) Adaptation of
Rhodococcus erythropolis DCL14 to growth on n-alkanes, alco-
hols and terpenes. Appl. Microbiol. Biotechnol. 67: 383-388.
35. de Carvalho, C., L. Wick, and H. Heipieper (2009) Cell wall
adaptations of planktonic and biofilm Rhodococcus erythropolis
cells to growth on C5 to C16 n-alkane hydrocarbons. Appl.
Microbiol. Biotechnol. 82: 311-320.
36. de Carvalho, C. C. C. R. and M. J. Caramujo (2008) Ancient pro-
cedures for the high-tech World: Health benefits and antimicro-
bial compounds from the Mediterranean empires. Open
Biotechnol. J. 2: 235-246.
37. Nair, M. K. M., J. Joy, P. Vasudevan, L. Hinckley, T. A. Hoag-
land, and K. S. Venkitanarayanan (2005) Antibacterial effect of
caprylic acid and monocaprylin on major bacterial mastitis
pathogens. J. Dairy Sci. 88: 3488-3495.
38. Kamdem, S. S., M. E. Guerzoni, J. Baranyi, and C. Pin (2008)
Effect of capric, lauric and [alpha]-linolenic acids on the division
time distributions of single cells of Staphylococcus aureus. Int. J.
Food Microbiol. 128: 122-128.
39. Dawson, P. L., G. D. Carl, J. C. Acton, and I. Y. Han (2002)
Effect of lauric acid and nisin-impregnated soy-based films on
the growth of Listeria monocytogenes on turkey bologna. Poult.
Sci. 81: 721-726.
40. Zheng, C. J., J. -S. Yoo, T. -G. Lee, H. -Y. Cho, Y. -H. Kim, and
W. -G. Kim (2005) Fatty acid synthesis is a target for antibacterial
activity of unsaturated fatty acids. FEBS Lett. 579: 5157-5162.
41. Yff, B. T. S., K. L. Lindsey, M. B. Taylor, D. G. Erasmus, and A.
K. Jäger (2002) The pharmacological screening of Pentanisia
prunelloides and the isolation of the antibacterial compound
palmitic acid. J. Ethnopharmacol. 79: 101-107.
42. Reid, K. A., A. K. Jäger, M. E. Light, D. A. Mulholland, and J. V.
Staden (2005) Phytochemical and pharmacological screening of
Sterculiaceae species and isolation of antibacterial compounds. J.
Ethnopharmacol. 97: 285-291.