Natural Product Research

Formerly Natural Product Letters

ISSN: 1478-6419 (Print) 1478-6427 (Online) Journal homepage: http://www.tandfonline.com/loi/gnpl20

Antibacterial and antifungal activities of the

polyphenolic fractions isolated from the seed coat
of Abrus precatorius and Caesalpinia crista

Lubna Mobin, Syed Asad Saeed, Rashida Ali, Syed Ghufran Saeed & Rahil
Ahmed

To cite this article: Lubna Mobin, Syed Asad Saeed, Rashida Ali, Syed Ghufran Saeed & Rahil
Ahmed (2018) Antibacterial and antifungal activities of the polyphenolic fractions isolated from
the seed coat of Abrus�precatorius and Caesalpinia�crista, Natural Product Research, 32:23,
2835-2839, DOI: 10.1080/14786419.2017.1378217

To link to this article: https://doi.org/10.1080/14786419.2017.1378217

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Published online: 26 Sep 2017.

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NATURAL PRODUCT RESEARCH, 2018
VOL. 32, NO. 23, 2835–2839
https://doi.org/10.1080/14786419.2017.1378217

SHORT COMMUNICATION

Antibacterial and antifungal activities of the polyphenolic

fractions isolated from the seed coat of Abrus precatorius and
Caesalpinia crista
Lubna Mobina, Syed Asad Saeeda, Rashida Alia,b, Syed Ghufran Saeeda and
Rahil Ahmedb
a
Department of Food Science and Technology, University of Karachi, Karachi, Pakistan; bQuality Assurance
Department, English Biscuit Manufacturers (Pvt.) Limited, Karachi, Pakistan

ABSTRACT ARTICLE HISTORY

Crude seed coat extracts from Abrus precatorius and Caesalpinia crista Received 29 June 2017
were purified into four different fractions namely phenolic acids, Accepted 29 August 2017
flavonols, flavanols and anthocyanin which were then examined
KEYWORDS
for their polyphenol contents and antimicrobial potentials. The Abrus precatorius; Caesalpinia
fractions derived from seed coat of A. precatorius were found more crista; seed coat; solid phase
potent with high phenolic and flavonoid contents as compared to extraction; polyphenol
C. crista fractions. The significant antibacterial activity was observed fractions; antimicrobial
against all strain tested by the fractions of both samples apart from
anthocyanin fraction. It was interesting to note that the phenolic
acid fractions of both samples was found more active against
gram-negative bacteria, while gram-positive bacteria were found
to be more sensitive towards flavonol fractions. The phenolic acid
and flavonol fractions being potent antibacterial were selected to
demonstrate the antifungal capacity of two samples. Among them,
phenolic acid fraction of both samples was found active towards all
the fungal strain.

1. Introduction
Today worldwide resistance to antimicrobials is becoming a serious problem. Thus, studies
in search of new natural antibiotics have always been in consideration in order to deal with

CONTACT  Lubna Mobin  lubna.mobin@hotmail.com, lubna.mobin@uok.edu.pk

 Supplemental data for this article can be accessed at https://doi.org/10.1080/14786419.2017.1378217.
© 2017 Informa UK Limited, trading as Taylor & Francis Group
2836 L. MOBIN ET AL.

this rising issue. The plants of Abrus precatorius and Caesalpinia crista are well-known for
their therapeutic values and antimicrobial activities of the different parts of both medicinal
plants have been explored (Bobbarala and Vadlapudi 2009; Ishan et al. 2013). However,
antimicrobial activity specifically from seed coat of these two medicinal plant has not been
investigated. The present research is therefore, based on the antimicrobial activity of the
polyphenol fractions viz. phenolic acid (Ia), flavanol (Ib), flavonol (Ic) and anthocyanin (II)
fractions derived from A. precatorius and C. crista seeds coat to evaluate the medicinal poten-
tial against certain diseases.

2. Result and discussion

The non-anthocyanin fractions (Ia, Ib and Ic) isolated from the crude extract of the A. preca-
torius and C. crista seeds coat have shown significant antibacterial activity towards all
screened species. Purification and isolation of different polyphenol compounds from crude
seed coat extract of A. precatorius and C. crista is schematically illustrated in Figure S1 (Siddiqi
et al. 2011). The identification of different class of polyphenolics in the isolated fractions I
and II (which is not the part of this manuscript) was carried out by the DAD-HPLC. The chro-
matographic conditions were set according to basic protocol described by Kim and Lee
(2002). The predominant phenolic compounds found in fraction I were epicatechin, syringic
acid, caeffic acid and vanillic acid, while delphinidin were the major anthocyanidin found in
the fraction II isolated from the A. precatorius seed coat (Table S1, Figures S2–S4). In the seed
coat of C. crista, among the taken 10 polyphenolic standards, no such polyphenolic com-
pounds were identified thus, the seed coat of C. crista is not supposed to be the main source
of polyphenols such as vanillic acid, syringic acid, caeffic acid, coumaric acid, epicatechin,
myrcetin, kaempherol, delphinidin, pelargonidin, cyanidin and their derivatives. The isolated
anthocyanin fraction (II) was found active only against S. aureus (Table 1). In both samples,
the order with respect to total phenolic and flavonoid contents was fraction II > fraction
Ic > fraction Ib > fraction Ia (Table S2), while the antibacterial effectiveness of fractions
derived from extracts were in sequence of fraction Ic > fraction Ib > fraction Ia for gram-pos-
itive bacteria and fraction Ia > fraction Ic > fraction Ib for gram-negative bacteria. The

Table 1. Antibacterial activity of the fractions isolated from crude extract of A. precatorius and C. crista
seed coat.a
Zone of inhibition (mm)
Seed coat
sample Frac S. typhi P. aerogenosa E.coli S. pyogenes S. aureus B. subtilis
A. precatorius Ia 45 ± 0.21e6 36 ± 0.17f5 28 ± 0.14e4 18 ± 0.25b2 22 ± 0.19c3 15 ± 0.25c1
Ib 25 ± 0.18c4 17 ± 0.18c2 15 ± 0.22b1 25 ± 0.20e4 33 ± 0.17 g5 22 ± 0.14e3
Ic 29 ± 0.15d3 25 ± 0.21e2 22 ± 0.25d1 35 ± 0.15f5 39 ± 0.13 h6 30 ± 0.18f4
II – – – 21 ± 0.91d1 25 ± 0.14e2 –
C. crista Ia 25 ± 0.90c6 22 ± 0.16d5 20 ± 0.19c4 15 ± 0.11a2 18 ± 0.12a3 10 ± 0.22a1
Ib 15 ± 0.19a3 12 ± 0.23a2 10 ± 0.99a1 20 ± 0.12c4 23 ± 0.11d5 12 ± 0.23b2
Ic 17 ± 0.22b3 13 ± 0.11b2 10 ± 0.15a1 20 ± 0.31c5 27 ± 0.14f6 19 ± 0.19d4
II – – – – 20 ± 0.11b –
Gentamycin 34 ± 0.12 30 ± 0.21 30 ± 0.14 27 ± 0.14 26 ± 0.11 32 ± 0.91
a
Values are mean of three replicates (5 mm well diameter).
(–) not observed.
No activity was observed in negative control.
Values in the same column with different superscript lowercase alphabets and values in the same row with superscript
lowercase numbers are significantly different at (p < 0.05).
 NATURAL PRODUCT RESEARCH 2837

fraction Ic, being higher in phenolic and flavonoids content, was found more active against
gram-positive bacteria with MIC between 31.25 and 250 µg/mL whereas gram-negative
bacteria were found more sensitive towards fraction Ia with MIC between 15.62 and 250 
µg/mL (Table S3). The genus S. aureus has shown the highest activity as gram-positive was
found more sensitive towards both the extracts, while S. typhi was the most susceptible
amongst the gram-negative species. The most resistant gram-positive bacteria against both
extract found was B. subtilis, while most resistant gram-negative strain was E.coli. The solvent
in which crude extracts were dissolved has shown no inhibitory effect. The polyphenol com-
pounds (flavonoids and non-flavonoids) basically exert antimicrobial action by causing
damage to the cytoplasmic membrane either by perforation causing reduction in the fluidity
of membrane (Tsuchiya and Iinuma 2000), preventing synthesis of nucleic acid (Mori et al.
1987) by inhibiting the enzyme topoisomerase (Plaper et al. 2003), preventing energy metab-
olism by inhibiting the activity of NADH cytochrome reductase (Haraguchi et al. 1998), inhib-
iting cell wall and cell membrane synthesis. The chelation of metal by phenolic compounds
is another possibility for the death of microorganisms (Negi 2012). The variations in the
fractions antibacterial potential could be explained on the basis of the fact that the inhibitory
action of polyphenol components against each microbe is different depending on the nature
of microbes and structure of polyphenols. In particular, the outer membrane complexity of
gram-negative bacteria acts as a protective barrier against lipophilic compounds. Thus,
higher antibacterial activities of fraction (Ia) of both the seed coat extracts against gram-neg-
ative bacteria was due to lesser lipophilic nature of phenolic acid. This increases the mem-
brane permeability and more uptake of phenolic acids (Ia) as compared to the flavanol (Ib)
and flavonol (Ic) in the cell that is responsible for successfully inhibiting the growth of
microbes by interfering with the electron transfer, reacting with nucleic acids and proteins
and/or other enzymatic activities (Dorman and Deans 2000). However, the inhibitory poten-
tial of flavanol and flavonol present in fraction Ib and Ic depends on the degree and position
of -OH on the A, B, C rings of these flavonoid (Lai and Roy 2004) and delocalised electron
system (Gochev et al. 2010). The absence of inhibitory effect of fraction II (except against
S.aureus) in both samples may be attributed to the membrane permeability resistance of
bacterial cell towards complex and polymerised structure of anthocyanin. These cause fewer
uptakes of antimicrobials inside the cell and thus, results in less and/or no inhibitory effect.
Fractions Ia and Ic of both samples, because of its higher antibacterial activity, were
selected to demonstrate the antifungal capability of A. precatorius and C. crista seed coat.
Results showed that both samples possessed antifungal property with A. precatorius seed
coat fractions (zone of inhibition ranged 25–35 mm) been more active than C. crista seed
coat (zone of inhibition ranged 21–30 mm). The phenolic acid fraction (Ia) of both samples
was found active towards all the tested fungal species, while flavonol fraction (Ic) has showed
activity only against C. albicans (Table 2). The polyphenol compound, depending on their
diverse structure, get diffuses through the fungal membrane and interfere with various
metabolic pathways by preventing the synthesis of protein, glucan, ergosterol, glocosamine
and chitin in fungi (Brul and Klis 1999).

3. Conclusion
In conclusion, it can be stated that the target sites of variety of flavonoids and non-flavonoids
on the microbial cells are different. Even the single class of polyphenolic compounds may
2838 L. MOBIN ET AL.

Table 2. Antifungal capacity of the Fraction Ia and Ib derived from crude extract of A. precatorius and
C. crista against different fungal strains.a
Zone of inhibition (mm)
A. precatorius C. crista Nystatin
Fungal species Fraction Ia Fraction Ic Fraction Ia Fraction Ic
C. albican 30 ± 0.09 18 ± 0.11 22 ± 0.14 15 ± 0.14 35 ± 0.15
A. flavus 35 ± 0.12 – 30 ± 0.90 – 40 ± 0.99
A. niger 32 ± 0.14 – 28 ± 0.13 – 37 ± 0.13
P. notatum 25 ± 0.12 – 21 ± 0.12 – 31 ± 0.18
a
Values are mean of three replicates (5 mm well diameter).
(–) not observed.
No activity was observed in negative control.

have different mode of actions. Results stated that high antibacterial activity of extracts or
fraction depends not only on the content of flavonoids and non-flavonoids but also the
chemical nature of the phenolics and flavonoids. The less lipophilic nature of non-flavonoid
polyphenol (phenolic acids) made them more reactive towards the outer membrane of
gram-negative bacteria, which eventually resulted in the loss of the functionality of cell.
Growth inhibition of Aspergillus species make them ideal to be use in food products for
minimising aflatoxin formation.

Disclosure statement
No potential conflict of interest was reported by the authors.

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