Saïd Ennahar

• University of Strasbourg

The compounds in leaf and stem extracts of Astragalus emarginatus Labill., a plant species used in traditional Lebanese medicine, were investigated for antioxidant properties. First, the activity of various extracts was assessed using the Trolox equivalent antioxidant capacity, oxygen radical absorption capacity, and 2,2-diphenyl-1-picryl-hydrazyl-...

Extracts from ‘Zhéri’ and ‘Hicaznar’ varieties of pomegranate, Punica granatum L., were obtained by subjecting powdered peels to extraction using water, water/ethanol (1:1; v/v), ethanol, acetone and heptane. Using the agar diffusion assay, extracts with water and/or ethanol were shown to display significant antimicrobial activity with diameters of...

A high-performance liquid chromatography-mass spectrometry technique hyphenated on-line with an immobilized enzyme reactor (IMER) was developed by the use of 3 known acetylcholinesterase (AChE) inhibitors (galanthamine, huperzine A and tacrine). This bioanalytical device allows qualitative comparison of the inhibitory strengths of AChE inhibitors....

Enterococcus faecium E16, a strain isolated from goats’ cheese made from unpasteurized milk, showed antibacterial activity, in particular against Listeria monocytogenes strains. The active compound was produced at high amounts in de Man Rogosa and Sharpe broth at 37 °C. It was purified to homogenity from culture supernatant by using a simple method...

The extraction of bioactive compounds from medicinal plants requires methods which are as diverse as the chemical nature of the compounds themselves. In this study, a 96-well microplate was used where solvent mixtures spanning wide ranges of selectivity and polarity were tested with the objective of extracting a broad range bioactive compounds from...

Aims: Oxidative stress (OS) plays a major role in type 2 diabetes and its vascular and hepatic complications, and novel therapeutic approaches include natural antioxidants. Our previous chemical and biological studies demonstrated the antioxidant activities of red cabbage (RC), and here, we aimed to determine the in vivo effects of 2-month long RC...

Background: Oxidative stress (OS) plays an important role in type 2 diabetes (T2D) pathogenesis and its complications. New therapies target natural antioxidants as an alternative and/or supplemental strategy to prevent and control them. Our previous chemical and biological studies highlighted the important antioxidant activities of cherries, among...

We analyzed the adhesion capacity to mucus of 31 Lactobacillus pentosus strains isolated from naturally fermented Aloreña green table olives using an immobilized mucin model. On the basis of their adhesive capacity to mucin, three phenotypes were selected for cell-wall protein proteomic analysis to pinpoint proteins involved in the adhesion process...

Acidity often prevents the undesirable microbial colonization both in fermented foods and under gastric conditions. Thus, the acid resistance of Lactobacillus pentosus strains used as starter cultures and/or probiotics requires further understanding. This was investigated by means of comparative proteomic approach using three strains representing t...

Background: Oxidative stress plays a major role in the development of several diseases. A healthy food diet, supplying the organism with antioxidants, is generally believed to help prevent this stress. Methods: With the aim of screening several antioxidants commonly found in food, a method using β pancreatic cells and based on the reduction of a...

Aims: The purpose of this study was to isolate, characterize and determine the structure and the antibacterial activities of a bacteriocin produced by Carnobacterium maltaromaticum CPN, a strain isolated from unpasteurized-milk Camembert cheese. Methods and results: This bacteriocin, termed maltaricin CPN, was produced at higher amounts in MRS b...

Probiotic bacterial cultures require resistance mechanisms to avoid stress-related responses under challenging environmental conditions; however, understanding these traits is required to discern their utility in fermentative food preparations, versus clinical and agricultural risk. Here, we compared the proteomic responses of Lb. pentosus MP-10, a...

Objectives: Oxidative stress plays a major role in the pathophysiology of type 2 diabetes and today new therapeutics target natural antioxidants as an alternative/additional strategy. Our studies, combining chemistry and biology, aimed to compare the anti-oxidant properties of fruits and vegetables (F&V) and to validate in vivo the effect of select...

Lactobacillus pentosus MP-10 isolated from brines of naturally fermented Alorena green table olives has potential probiotic traits includin inhibition of human pathogenic bacteria, survival at low ph (1.5) and bile salt tolerance (3%). Lactobacillus pentosus MP-10 was intrinsically resistant to some antibiotics but sensitive to different biocides i...

This study investigates the mechanisms of UV-A (315 to 400 nm) photocatalysis with titanium dioxide (TiO2) applied to the degradation of Escherichia coli and their effects on two key cellular components: lipids and proteins. The impact of TiO2 photocatalysis on E. coli survival was monitored by counting on agar plate and by assessing lipid peroxida...

In this study, the thermal stability of phospholipids (PLs) extracted from coffee beans was investigated. Chemical analysis was used to obtain information about the effect of heat treatment on the content of PLs in roasted coffee beans. Normal phase high-performance liquid chromatography (HPLC) combined with evaporative light scattering detector (E...

DNA damage in frozen and chilled salmon (Salmo salar) was investigated using the comet assay. Frozen salmon was thawed at 10 �C and chilled salmon was kept in flake ice, mimicking a thermal abuse and an extended storage, respectively. Electrophoresis conditions were as follows: a second layer of lowmelting- point agarose at 1 g/100 mL; a voltage of...

To investigate cell determinants of resistance to gastric acidity in Lactobacillus plantarum using comparative proteomics. Among ten L. plantarum strains that were tested for their acid resistance in vitro, three strains with different phenotypes were selected for comparative proteomic analysis: LC 804 (resistant), CIP A159 (intermediate) and CECT...

Several SPE sorbents were investigated for the extraction of a group of chemically diverse isothiocyanates (ITCs). They included bonded silica, carbon-based, and polymer-based sorbents with various functional groups. Results showed large differences in the ability of these sorbents to simultaneously extract ITCs from standard solutions. Recovery ra...

The largely adopted reversed-phase HPLC analysis of the molecular species of isothiocyanates (ITCs) was performed and showed losses during the chromatographic run with eight ITCs. These losses, which obviously impact the accuracy of quantitative determinations, were due to precipitation in the chromatographic system. At 22°C, they ranged from 5.4%...

The antimicrobial activities of ten isothiocyanates (ITCs), namely sulforaphane, iberin, allyl-, benzyl-, methyl-, phenyl-, phenylethyl-, propyl-, 3-methylthiophenyl-, and 3-methylthiopropyl-ITC, were tested against a range of pathogenic and food spoilage bacteria. The activities of six ITCs were investigated for the first time. The growth of the b...

A liquid chromatographic-electrospray ionization-tandem mass spectrometric (LC-ESI-MS(2)) method has been developed for determination of the molecular species of phosphatidylethanolamine (PE) in four food matrices (soy, egg yolk, ox liver, and krill oil). The extraction and purification method consisted of a pressurized liquid extraction procedure...

The identification of cell determinants involved in probiotic features is a challenge in current probiotic research. In this work, markers of bile tolerance in Lactobacillus casei were investigated using comparative proteomics. Six L. casei strains were classified on the basis of their ability to grow in the presence of bile salts in vitro. Constit...

In this study, molecular species of sphingomyelin (SM) in egg yolk, calf brain, ox liver, and krill oil were investigated. Classes of phospholipids (PLs) were purified, identified, and quantified by normal phase semipreparative high-performance liquid chromatography (HPLC) combined with evaporative light scattering detectors (ELSD). For SM molecula...

Numerous isothiocyanates (ITCs) are poorly soluble in water which causes their precipitation in aqueous mobile phases used in reversed phase liquid chromatography (RP-LC), thus impacting the accuracy of the quantification. By comparing the amounts of ITCs injected and released from the column, losses could be estimated at 5-32% depending on polarit...

With the aim of discovering new natural active extracts against malaria parasites, Icacina senegalensis was selected after an ethnopharmacological survey conducted on plants used in traditional malaria treatment in Senegal. Different concentrations of the plant extract and fractions were tested on synchronized Plasmodium falciparum cultures at the...

Identification of differentially expressed protein spots among L. plantarum LC 56, LC 804 and 299 V in standard growth conditions. The table lists proteins with at least a twofold difference of expression (p-value < 0.05) between the three strains cultured in MRSC. Identification was achieved following excision of differentially expressed spots bet...

Lactic acid bacteria are commonly marketed as probiotics based on their putative or proven health-promoting effects. These effects are known to be strain specific but the underlying molecular mechanisms remain poorly understood. Therefore, unravelling the determinants behind probiotic features is of particular interest since it would help select st...

The extraction of three major phospholipid (PL) classes contained in soybean, egg yolk, calf brain, and ox liver was investigated by means of two methods. The PL amounts were evaluated. A new method, based on pressurized liquid extraction (PLE), was applied for total lipids (TL), including PL, extraction and compared with a standard liquid extracti...

Heat-induced oxidative modifications of two phosphatidylcholine molecular species as potential functional food components were evaluated. 1-stearoyl-2-oleoyl-sn-glycero-3-phosphocholine (SOPC) and 1-stearoyl-2-linoleoyl-sn-glycero-3-phosphocholine (SLPC) were chosen as models. The optimal temperature for hydroperoxide formation was determined by MS...

An investigation was carried out into the stability of fatty acyl groups to heat-induced oxidative changes as affected by their chemical environment. The behavior of oleic and linoleic acyl groups when esterified in triacylglycerols (TAGs) and phosphatidylcholines (PCs) was evaluated. The monitoring of the oxidative degradation using liquid chromat...

This study is a contribution to the exploration of natural phospholipid (PL) sources rich in long-chain polyunsaturated fatty acids (LC-PUFAs) with nutritional interest. Phosphatidylcholines (PCs) were purified from total lipid extracts of different food matrices, and their molecular species were separated and identified by liquid chromatography-el...

Enterococcus faecalis WHE 96, a strain isolated from soft cheese based on its anti-Listeria activity, produced a 5,494-Da bacteriocin that was purified to homogeneity by ultrafiltration and cation-exchange and reversed-phase chromatographies. The amino acid sequence of this bacteriocin, named enterocin 96, was determined by Edman degradation, and i...

Phytosteryl esters (PE) are used as ingredients in functional food to decrease plasma concentration of low density lipoprotein-cholesterol (LDL-C). Effective impairment of cholesterol absorption by PE suggests that these esters are hydrolyzed by the pancreatic cholesterol esterase (CEase, EC 3.1.1.13) and the liberated sterol may interfere with cho...

Enterococcus faecium IT62, a strain isolated from ryegrass in Japan, produces three bacteriocins (enterocins L50A, L50B, and IT) that have been previously purified and the primary structures of which have been determined by amino acid sequencing (E. Izquierdo, A. Bednarczyk, C. Schaeffer, Y. Cai, E. Marchioni, A. Van Dorsselaer, and S. Ennahar, Ant...

Enterococcus faecium WHE 81, a multi-bacteriocin producer, was tested for its antimicrobial activity on Listeria monocytogenes in Munster cheese, a red smear soft cheese. The naturally delayed and superficial contamination of this type of cheese allowed the use of E. faecium WHE 81 at the beginning of the ripening as a surface culture. A brine solu...

The identification of cell components involved in probiotic activities is a challenge in current probiotic research. In this work, a new approach based on proteomics as an analytical tool for the identification of characteristic protein profiles related to adhesion to mucin as a model probiotic property was used. Three Lactobacillus plantarum strai...

Whey proteins were used as a coating material to improve encapsulation of Lactobacillus plantarum strains in calcium alginate beads. L. plantarum 299v, L. plantarum 800 and L. plantarum CIP A159 were used in this study. Inactivation experiments were carried out in simulated gastric fluid (SGF) and simulated intestinal fluid (SIF). Cross-sections of...

Enterococcus faecium IT62, isolated from ryegrass in Japan, was shown to produce three different bacteriocins, two of which had molecular masses and amino acid sequences that corresponded to those of enterocin L50A and enterocin L50B. These peptides existed, however, as chemically modified forms that were either N formylated or N formylated and oxi...

Eight Bifidobacterium longum strains, including reported probiotic strains (commercial and noncommercial), collection strains, and laboratory isolates, were investigated for their ability to adhere to mucin as well as their ability to tolerate acid and bile. Strains could be discriminated based on their sensitivity at pH values of 2.0 to 2.5 and bi...

Modulation of host immunity is one of the proposed benefits of the consumption of probiotics. Nonetheless, comparative studies on the immunological properties that support the selection of strains of the same species for specific health benefits are limited. In this study, the ability of different strains of Bifidobacterium longum to induce cytokin...

Soft and red smear cheeses are frequently contaminated by Listeria monocytogenes, sometimes at relatively high concentration (< 105 CFUg-1). This bacterium is radiosensitive (D10 value of approximately 0.45 kGy) but irradiation of the whole cheese by X-rays induced off-flavours when the dose exceeded 1.0 kGy. Irradiation could be effective in elimi...

UV radiation is able to induce lipid peroxidation. Photooxidation-induced beta-sitosterol oxides were monitored in four vegetable oils exposed to sunlight for 10, 20, and 30 days during May 2005 (northeastern France), exposed to artificial light generated by a high-pressure Hg lamp for 21, 42, and 63 h at room temperature, and exposed to a 10 MeV e...

In the last decade, a variety of ribosomally synthesized antimicrobial peptides or bacteriocins produced by lactic acid bacteria have been identified and characterized. As a result of these studies, insight has been gained into fundamental aspects of biology and biochemistry such as producer self protection, membrane–protein interactions, and prote...

Effects of abusive storage conditions on the quality of fresh chicken were studied by detecting DNA damage to breast fillets and liver with the neutral comet assay. Chilled samples were kept at 4 degrees C for prolonged periods, whereas frozen samples were exposed to temperatures of 4 degrees C, representing inadvertent thawing, and 20 degrees C, r...

Phylogenetic analysis of 16S rRNA gene sequences revealed that Enterococcus solitarius is not a member of the genus Enterococcus, but is related to species of the genus Tetragenococcus. On a phylogenetic tree, E. solitarius clustered with Tetragenococcus halophilus and Tetragenococcus muriaticus, with which it showed the highest 16S rRNA gene seque...

The taxonomic positions of Weissella kimchii and Weissella cibaria were examined using phenotypic tests, 16S rRNA gene sequencing and DNA-DNA hybridization. The two species displayed very similar sugar-fermentation patterns and produced the D-form of lactic acid. Phylogenetic analysis showed that W. cibaria is the closest neighbour of W. kimchii, s...

A total of 161 low-G+C-content gram-positive bacteria isolated from whole-crop paddy rice silage were classified and subjected to phenotypic and genetic analyses. Based on morphological and biochemical characters, these presumptive lactic acid bacterium (LAB) isolates were divided into 10 groups that included members of the genera Enterococcus, Lac...

Enterococcus faecium WHE 81, isolated from cheese, has been reported to produce a bacteriocin called "enterocin 81" [J. Appl. Microbiol. 85 (1998) 521.]. Purification of "enterocin 81" was carried out using ammonium sulfate precipitation, desalting on ODP-90 reverse-phase column, and purification through SP Sepharose HP cation exchange and C2/C18 r...

Thirty-one Listeria strains were tested for sensitivity to four class IIa bacteriocins, namely, enterocin A, mesentericin Y105, divercin V41, and pediocin AcH, and to nisin A. Class IIa bacteriocins displayed surprisingly similar antimicrobial patterns ranging from highly susceptible to fully resistant strains, whereas nisin A showed a different pa...

Enterocin A produced by Enterococcus faecium EFM01 displayed a narrow antimicrobial spectrum, mainly directed against Listeria spp. In particular, the bacteriocin was extremely active against 13 Listeria monocytogenes strains. This high specificity of action of enterocin A for Listeria spp. relative to lactic acid bacteria, together with its broad...

Enterococcus faecium WHE 81, isolated from a smear-surface soft cheese, was shown to produce at least two bacteriocins, with high anti-Listeria activity. One of the bacteriocins produced was purified to homogeneity by ammonium sulfate precipitation, desalting on an ODP-90 reversephasecolumn, SP Sepharose HP cation exchange chromatography and C22/C1...

In the last decade, a variety of ribosomally synthesized antimicrobial peptides, or bacteriocins, produced by lactic acid bacteria have been identified and characterized. As a result of these studies, insight has been gained into various fundamental aspects of biology and biochemistry such as bacteriocin processing and secretion, mechanisms of cell...

Lactic acid bacteria, Chizuka, Yasaka 5B,7B,8B, and 9B, which have been previously isolated from different well-aged Nukadoko in the Northern Kyushu area, were investigated for bacteriocin production and purification. Their isolates were shown to produce bacteriocin-like substances. However, the properties of these substances were surprisingly iden...

Enterocin 81, a bacteriocin produced by Enterococcus faecium WHE 81 previously isolated from cheese, exhibited a very narrow spectrum of activity, which is mainly directed against enterococci and Listeria spp. including Listeria monocytogenes. Enterocin 81 activity, which was extremely rapid with maximal effect achieved within 30 min, could not be...

The anti-Listeria monocytogenes activity of Lactobacillus plantarum WHE 92, a pediocin AcH producer, was investigated in Munster cheese, a smear-surface soft cheese. The appearance of L. monocytogenes in the cheese, which naturally occurs solely in the crust and never before 1 week of ripening, could be prevented by spraying a cell suspension of L....

Among 1,962 bacterial isolates from a smear-surface soft cheese (Munster cheese) screened for activity against Listeria monocytogenes, six produced antilisterial compounds other than organic acids. The bacterial strain WHE 92, which displayed the strongest antilisterial effect, was identified at the DNA level as Lactobacillus plantarum. The protein...

Amicrobiologicalstudyofsmear-surfacesoftcheeses(Mun- ster type), carried out in 1992 with 357 samples prepared from pasteurized milk and 115 samples prepared from raw milk, has shown high-level contamination of these cheeses withListeria monocytogenes. This bacterium was found in almost 60% of cheeses prepared from pasteurized milk (with levels hig...