Ahmed O. El-Gendy

Nanotechnology: A Valuable Strategy to Improve Bacteriocin Formulations

Bacteriocins are proteinaceous antibacterial compounds, produced by diverse bacteria, which have been successfully used as: (i) food biopreservative; (ii) anti-biofilm agents; and (iii) additives or alternatives to the currently existing antibiotics, to minimize the risk of emergence of resistant strains. However, there are several limitations that challenge the use of bacteriocins as biopreservatives/antibacterial agents. One of the most promising avenues to overcome these limitations is the use of nanoformulations. This review highlights the practical difficulties with using bacteriocins to control pathogenic microorganisms, and provides an overview on the role of nanotechnology in improving the antimicrobial activity and the physicochemical properties of these peptides.

Exploring the Antimicrobial and Antitumor Potentials of Streptomyces sp. AGM12-1 Isolated from Egyptian Soil

The occurrence of extensive antibiotics resistant bacteria increased the demands for mining out new sources of antimicrobial agents. Actinomycetes, especially Streptomyces sp. have grasped considerable attention worldwide due to production of many useful bioactive metabolites. In the present study, a total of 52 actinomycetes were isolated from agricultural soil samples in Beni-Suef, Egypt. All isolates were characterized based on colony morphology, mycelium coloration, and pigment diffusion. They were screened for their capabilities to show antimicrobial activities against different indicator microorganisms, and only 20 isolates have shown significant antimicrobial activities against at least one of the tested indicator microorganisms. The isolate AGM12-1 was active against all tested microorganisms and showed a marked antitumor activity with IC50 3.3 and 1.1 μg/ml against HCT-116 and HepG-2 cell lines respectively. It was genotypically characterized as Streptomyces sp. with the presence of PKS Π biosynthetic gene cluster. Mannitol, ammonium sulfate, pH 7, 2% inoculum size and incubation for 11 days at 30°C were the optimum conditions that used to maximize the production and hence allowed purification of one active antimicrobial compound to homogeneity using high performance liquid chromatography with a molecular mass of m/z 488.05. Nuclear magnetic resonance structural elucidation showed that this compound was a diketopiperazine derivative.

The fsr Quorum-Sensing System and Cognate Gelatinase Orchestrate the Expression and Processing of Proprotein EF_1097 into the Mature Antimicrobial Peptide Enterocin O16

UNLABELLED A novel antimicrobial peptide designated enterocin O16 was purified from Enterococcus faecalis. Mass spectrometry showed a monoisotopic mass of 7,231 Da, and N-terminal Edman degradation identified a 29-amino-acid sequence corresponding to residues 90 to 119 of the EF_1097 protein. Bioinformatic analysis showed that enterocin O16 is composed of the 68 most C-terminal residues of the EF_1097 protein. Introduction of an in-frame isogenic deletion in the ef1097 gene abolished the production of enterocin O16. Enterocin O16 has a narrow inhibitory spectrum, as it inhibits mostly lactobacilli. Apparently, E. faecalis is intrinsically resistant to the antimicrobial peptide, as no immunity connected to the production of enterocin O16 could be identified. ef1097 has previously been identified as one of three loci regulated by the fsr quorum-sensing system. The introduction of a nonsense mutation into fsrB consistently impaired enterocin O16 production, but externally added gelatinase biosynthesis-activating pheromone restored the antimicrobial activity. Functional genetic analysis showed that the EF_1097 proprotein is processed extracellularly into enterocin O16 by the metalloprotease GelE. Thus, it is evident that the fsr quorum-sensing system constitutes the regulatory unit that controls the expression of the EF_1097 precursor protein and the protease GelE and that the latter is required for the formation of enterocin O16. On the basis of these results, this study identified antibacterial antagonism as a novel aspect related to the function of fsr and provides a rationale for why ef1097 is part of the fsr regulon. IMPORTANCE The fsr quorum-sensing system modulates important physiological functions in E. faecalis via the activity of GelE. The present study presents a new facet of fsr signaling. The system controls the expression of three primary target operons (fsrABCD, gelE-sprE, and ef1097-ef1097b). We demonstrate that the concerted expression of these operons constitutes the elements necessary for the production of a bacteriocin-type peptide and that antimicrobial antagonism is an intrinsic function of fsr. The bacteriocin enterocin O16 consists of the 68 most C-terminal residues of the EF_1097 secreted proprotein. The GelE protease processes the EF_1097 proprotein into enterocin O16. In this manner, fsr signaling enables E. faecalis populations to express antimicrobial activity in a cell density-dependent manner.

Clinical Screening for Bacteriocinogenic Enterococcus faecalis Isolated from Intensive Care Unit Inpatient in Egypt

A bacterium originally isolated from stool sample from ICU inpatient was biochemically and molecularly characterized and identified as Enterococcus faecalis OS6. This strain showed the ability to exert antimicrobial activities against some members of Gram-positive bacteria as Lactobacilli and Enterococci. However, no activities were detected against all tested Gram-negative and fungal indicator strains. Diminishing of antimicrobial activities upon heat and proteinase K treatments confirmed the proteinaceous nature of the recorded activity. Therefore, strain OS6 was extensively screened for the presence of 10 common bacteriocin structural genes where genes of Enterolysin A and Cytolysin were detected and confirmed by further gene sequencing. Further characterization of strain OS6 showed several virulence determinants including gelatinase, haemolysin, bile salt hydrolase and multiple antibiotic resistance traits towards 17 out of 31 different antibiotics. Most of these 17 antibiotics belonged to Cephalosporins, Aminoglycosides, Lincomycins, Polypeptides, Quinolones, Rifamycins and Sulfonamides classes. To the best of our knowledge, this is the first attempt to report production of bacteriocins (mainly Enterolysin A) in between pathogenic E. faecalis isolated from human clinical specimen in Egypt.

Enhancement of the productivity of the potent bacteriocin avicin A and improvement of its stability using nanotechnology approaches.

Herein, enhancements of the yield and antimicrobial activity duration of the bacteriocin avicin A were accomplished using fractional factorial design (FFD) and layered double hydroxide (LDH) nanoparticles. Firstly, potential factors affecting bacteriocin production were selected for preliminary study. By a 25-1 FFD, high pH was shown to have a positive effect on avicin A yield, while temperature and duration of incubation, as well as peptone nitrogen sources all had negative effects. The highest bacteriocin production and activity (2560 BU/ml) were observed after 30 h of incubation at 30 °C, with pH adjustment at 7, and in the presence of 2 g mannitol as carbon source and 2.2 g peptone as nitrogen source. Secondly, avicin A nanocomposites with different LDH precursors were tested. Only avicin A-ZnAl-CO3 LDH demonstrated a potent antimicrobial activity against Lactobacillus sakei LMGT 2313 that lasted for at least 24 days, as compared to the values of 6 and 15 days observed with the free avicin A that has been stored at room temperature and at 4 °C, respectively. In conclusion, avicin A production and stability can be improved by manipulating the growth conditions and media composition, together with conjugation to LDHs.

Microbial diversity of mer operon genes and their potential rules in mercury bioremediation and resistance

REVIEW ARTICLE Microbial Diversity of Mer Operon Genes and Their Potential Rules in Mercury Bioremediation and Resistance Martha M. Naguib, Ahmed O. El-Gendy and Ahmed S. Khairalla Department of Biotechnology and Life Sciences, Faculty of Post Graduate Studies for Advanced Sciences, Beni-Suef University, Beni-Suef 62511, Egypt Department of Microbiology and Immunology, Faculty of Pharmacy, Beni-Suef University, Beni-Suef 62511, Egypt

Characterization of natural bioactive compounds produced by isolated bacteria from compost of aromatic plants

This study aimed to highlight the importance of compost from aromatic plants as a stunning source for several bio active compounds generated from their inhabited thermophilic bacteria. Some of the isolated compounds could have a potential role in the treatment of microbial infections.