Volume 2 • Issue 6 • 1000131
Clin Microbial
ISSN: 2327-5073 CMO, an open access journal
Open Access
Ahmed et al., Clin Microbial 2013, 2:6
DOI: 10.4172/2327-5073.1000131
Open Access
Clinical Microbiology: Open Access
The Relationship between Bioactive Compounds with Diastase Activity and
Antibacterial Synergy of Honey and Potato Starch Combinations against
Klebsiella pneumonia
Moussa Ahmed*, Baghdad Khiati, Saad Aissat Noureddine Djebli, Abdelmalek Meslem and Salima Bacha
Pharmacognosy and Api-Phytotherapy Research Laboratory, Mostaganem University, Algeria
Keywords: Algerian honey; Antibacterial activity; Potato starch;
Synergism; Bioactive compounds; Diastase activity
Introduction
Klebsiella pneumoniae, an important human pathogen, is the most
common cause for pneumonia, bacteremia, and septicemia, is also
involved in urinary tract infections (UTI) [1] Treatment of K. pneumoniae
infections has become increasingly difcult because of the predominance
of multiple-antibiotic-resistant strains [2,3]. Antibiotic resistance has
increased substantially in recent years and is posing an ever-increasing
therapeutic problem [4,5]. Honey is a natural, nontoxic, and inexpensive
product for the need of novel therapies against bacterial infections. Te
clinical use of honey has an enormous potential, especially in the fght
against antibiotic-resistant strains [5,6]. Several bioactive compounds
have been identifed in honey which contributed to its antibacterial
action. Te main factor is the high osmotic activity of honey, which does
not allow bacterial growth [7]. Recent experimental fnding indicated
that the amylase present in honey increases the osmotic efect in the
media by increasing the amount of sugars and consequently increasing
the antibacterial activity [8]. Honey contains small amounts of diferent
enzymes, notably, diastase (α-and β -amylase), invertase (α-glucosidase),
glucose-oxidase, catalase and acid phosphatase, which come from
nectar sources, salivary fuids and the pharyngeal gland secretions of
the honeybee [9]. Diastase is a natural enzyme of honey. Αlpha-amylase
degrades starch to a mixture of the disaccharide maltose, the trisaccharide
maltotriose and oligosaccharides known as dextrin’s [10]. Also phenolic
compounds may contribute to antimicrobial activity [11]. Te inhibitory
efects of polyphenols for α-amylases have attracted great interest among
researchers [12,13]. In this study, we evaluated the antibacterial activity
of 6 varieties of honey against K. pneumoniae and their combination
with potato starch in correlation with diastase number (α- amylase), and
Bioactive Compounds (total favonoid and polyphenol content).
Materials and Methods
Honey samples
Six (n=6) commercial honeys of diferent foral sources and
*Corresponding author: Moussa Ahmed, Pharmacognosy and Api-Phytotherapy
Research Laboratory, Mostaganem University, Algeria, Tel: +213 65234059; Fax:
+213 65234059; E-mail: moussa7014@yahoo.fr
Received July 22, 2013; Accepted October 17, 2013; Published October 22,
2013
Citation: Ahmed M, Khiati B, Aissat S, Djebli N, Meslem A, et al. (2013) The
Relationship between Bioactive Compounds with Diastase Activity and Antibacterial
Synergy of Honey and Potato Starch Combinations against Klebsiella pneumonia.
Clin Microbial 2: 131. doi: 10.4172/2327-5073.1000131
Copyright: © 2013 Ahmed M, et al. This is an open-access article distributed under
the terms of the Creative Commons Attribution License, which permits unrestricted
use, distribution, and reproduction in any medium, provided the original author and
source are credited.
Abstract
Honey samples produced by Apismellifera, both uniforal and multiforal from different sources in Algeria were
examined for their antibacterial capacity and their synergism with potato starch. An agar incorporation technique was
used to assess the minimum inhibition concentration (MIC) and the minimum inhibition additive concentration (MIAC)
of honey against K. pneumoniae ATCC 27736. The physicochemical properties, α-amylases activity, total phenolic
and favonoid content, of six representative honey samples were determined. The MIC for the six varieties of honey
without potato starch against K. pneumoniae ranged between 14% and 24% (v ⁄ v). When starch was incubated
with honey and then added to media, an MIC drop was noticed with each variety and it ranged between 5.55 % and
16.66%.The total phenolic content of honey samples was between 1.50–108.21 mg GAE/100 g honey as gallic acid
equivalent, total favonoid content varied from 5.41 to 9.94 mg Catechin/kg . Mean value for diastase was 16.55 ± 2.8
(range 7.3–23.5) expressed as diastase number in Gothe’sscale. No signifcant correlation was established between
α-amylases activity and bioactive compounds. Honey: potato starch combinations show real potential for future use
as a treatment for infections caused by K. pneumonia.
geographical origins where purchased from local market and lef at
room temperature until further analysis.
Preparation of the stock starch solution
Te stock starch solution was prepared by dissolving 0.5 g of dried
soluble starch in deionised water in a volumetric fask. Afer heating
and stirring the solution for approximately ten minutes, starch was
completely dissolved, and the volumetric fask was flled with deionised
water to the mark.
Bacterial culture and inoculum preparation
Pure culture of K. pneumonia 27736 was obtained from the
Department of Biology, Faculty of Sciences, Mostaganem University,
Algeria. Te bacteria was grown on Nutrient Agar (NA; Merck
Germany) slant, incubated at 37°C for 24 h, and kept at 4°C until further
use. Bacterial suspension was prepared by inoculating one loopful of
the 24-h-old bacterial colonies into 10.0 ml of sterilized distilled water.
Te inoculums size was adjusted to match the turbidity of McFarland
0.5 scale (1×10
8
cells/ml) and diluted with sterilized distilled water to
the inoculums size of 1×10
7
cells/ml.
Diastase activity (Diastase number)
Diastase activity was measured with Phadebas, according to the
Harmonized Methods of the European Commission of Honey [14].
C
l
i
n
i
c
a
l
M
i
c
r
o
b
i
o
l
o
g
y
:
O
p
e
n
A
c
c
e
s
s
ISSN: 2327-5073
Editorial