scholarly journals Addition of L-cysteine to the N- or C-terminus of the all-D-enantiomer [D(KLAKLAK)2] increases antimicrobial activities against multidrug-resistant Pseudomonas aeruginosa, Acinetobacter baumannii and Escherichia coli

PeerJ ◽  
2020 ◽  
Vol 8 ◽  
pp. e10176
Author(s):  
Maki K. Ohno ◽  
Teruo Kirikae ◽  
Eisaku Yoshihara ◽  
Fumiko Kirikae ◽  
Isao Ishida

Background Antimicrobial peptides have a broad spectrum of antimicrobial activities and are attracting attention as promising next-generation antibiotics against multidrug-resistant (MDR) bacteria. The all-d-enantiomer [D(KLAKLAK)2] has been reported to have antimicrobial activity against Escherichia coli and Pseudomonas aeruginosa, and to be resistant to protein degradation in bacteria because it is composed of D-enantiomer compounds. In this study, we demonstrated that modification of [D(KLAKLAK)2] by the addition of an L-cysteine residue to its N- or C- terminus markedly enhanced its antimicrobial activities against Gram-negative bacteria such as MDR Acinetobacter baumannii, E. coli, and P. aeruginosa. Methods The peptides [D(KLAKLAK)2] (DP), DP to which L-cysteine was added at the N-terminus C-DP, and DP to which L-cysteine was added at the C-terminus DP-C, were synthesized at >95% purity. The minimum inhibitory concentrations of peptides and antibiotics were determined by the broth microdilution method. The synergistic effects of the peptides and the antibiotics against MDR P. aeruginosa were evaluated using the checkerboard dilution method. In order to assess how these peptides affect the survival of human cells, cell viability was determined using a Cell Counting Kit-8. Results C-DP and DP-C enhanced the antimicrobial activities of the peptide against MDR Gram-negative bacteria, including A. baumannii, E. coli, and P. aeruginosa. The antimicrobial activity of DP-C was greater than that of C-DP, with these peptides also having antimicrobial activity against drug-susceptible P. aeruginosa and drug-resistant P. aeruginosa overexpressing the efflux pump components. C-DP and DP-C also showed antimicrobial activity against colistin-resistant E. coli harboring mcr-1, which encodes a lipid A modifying enzyme. DP-C showed synergistic antimicrobial activity against MDR P. aeruginosa when combined with colistin. The LD50 of DP-C against a human cell line HepG2 was six times higher than the MIC of DP-C against MDR P. aeruginosa. The LD50 of DP-C was not altered by incubation with low-dose colistin. Conclusion Attachment of an L-cysteine residue to the N- or C-terminus of [D(KLAKLAK)2] enhanced its antimicrobial activity against A. baumannii, E. coli, and P. aeruginosa. The combination of C-DP or DP-C and colistin had synergistic effects against MDR P. aeruginosa. In addition, DP-C and C-DP showed much stronger antimicrobial activity against MDR A. baumannii and E. coli than against P. aeruginosa.

mBio ◽  
2018 ◽  
Vol 9 (4) ◽  
Author(s):  
Yingbo Shen ◽  
Zuowei Wu ◽  
Yang Wang ◽  
Rong Zhang ◽  
Hong-Wei Zhou ◽  
...  

ABSTRACTThe recent emergence of a transferable colistin resistance mechanism, MCR-1, has gained global attention because of its threat to clinical treatment of infections caused by multidrug-resistant Gram-negative bacteria. However, the possible transmission route ofmcr-1amongEnterobacteriaceaespecies in clinical settings is largely unknown. Here, we present a comprehensive genomic analysis ofEscherichia coliisolates collected in a hospital in Hangzhou, China. We found thatmcr-1-carrying isolates from clinical infections and feces of inpatients and healthy volunteers were genetically diverse and were not closely related phylogenetically, suggesting that clonal expansion is not involved in the spread ofmcr-1. Themcr-1gene was found on either chromosomes or plasmids, but in most of theE. coliisolates,mcr-1was carried on plasmids. The genetic context of the plasmids showed considerable diversity as evidenced by the different functional insertion sequence (IS) elements, toxin-antitoxin (TA) systems, heavy metal resistance determinants, and Rep proteins of broad-host-range plasmids. Additionally, the genomic analysis revealed nosocomial transmission ofmcr-1and the coexistence ofmcr-1with other genes encoding β-lactamases and fluoroquinolone resistance in theE. coliisolates. These findings indicate thatmcr-1is heterogeneously disseminated in both commensal and pathogenic strains ofE. coli, suggest the high flexibility of this gene in its association with diverse genetic backgrounds of the hosts, and provide new insights into the genome epidemiology ofmcr-1among hospital-associatedE. colistrains.IMPORTANCEColistin represents one of the very few available drugs for treating infections caused by extensively multidrug-resistant Gram-negative bacteria. The recently emergentmcr-1colistin resistance gene threatens the clinical utility of colistin and has gained global attention. Howmcr-1spreads in hospital settings remains unknown and was investigated by whole-genome sequencing ofmcr-1-carryingEscherichia coliin this study. The findings revealed extraordinary flexibility ofmcr-1in its spread among genetically diverseE. colihosts and plasmids, nosocomial transmission ofmcr-1-carryingE. coli, and the continuous emergence of novel Inc types of plasmids carryingmcr-1and newmcr-1variants. Additionally,mcr-1was found to be frequently associated with other genes encoding β-lactams and fluoroquinolone resistance. These findings provide important information on the transmission and epidemiology ofmcr-1and are of significant public health importance as the information is expected to facilitate the control of this significant antibiotic resistance threat.


Author(s):  
Samaila Abubakar ◽  
Musa Muktari ◽  
Rejoice Atiko

The synthesis and antimicrobial application of Co (III) and Fe (III) complexes of imine functionalized N-heterocyclic carbene (Imino-NHC) ligands is reported. The ligand precursors 1-(2-[(hydroxyl-benzylidene)-amino]-ethyl)-3-R-3H-imidazol-1-ium bromide where R = pyridyl (1a) and benzyl (1b) have been reported in our previous work. The in-situ generated ligands of 1a and 1b have been successfully coordinated to CoBr2 and [FeI(Cp)(CO)2] leading to the isolation of air-stable N^C^N^O four coordinate Co(III)  complex 2 and a six-coordinate Fe(III) complex 3. The synthesised complexes were both found to be NMR inactive hence were characterize using FTIR and LRMS. The complexes were screened for antimicrobial activities against four gram-negative bacteria Escherichia Coli (E-coli), Shigella, Klebsiella pneumoniae (K. Pneumoniae) and Salmonella typhi (S. typhi) and a gram positive bacteria Staphylocossus aureus (S. aureus). The antimicrobial test was conducted using disc diffusion methods and based on the concentrations of 100, 200, 300, 400 and 500 µg/ mL, significant activities were recorded for both cobalt and the iron complexes.


2013 ◽  
Vol 2013 ◽  
pp. 1-7 ◽  
Author(s):  
Paulina L. Páez ◽  
Claudia M. Bazán ◽  
María E. Bongiovanni ◽  
Judith Toneatto ◽  
Inés Albesa ◽  
...  

The prevalence of antibiotic resistance has resulted in the need for new approaches to be developed to combat previously easily treatable infections. The main aim of this work was to establish the potential of the syntheticα-diimine chromium(III) and ruthenium(II) complexes (where theα-diimine ligands are bpy = 2,2-bipyridine, phen = 1,10-phenanthroline, and dppz = dipyrido[3,2-a:2′,3′-c]-phenazine) like [Cr(phen)3]3+, [Cr(phen)2(dppz)]3+, [Ru(phen)3]2+, and [Ru(bpy)3]2+as antibacterial agents by generating oxidative stress. The [Cr(phen)3]3+and [Cr(phen)2(dppz)]3+complexes showed activity against Gram positive and Gram negative bacteria with minimum inhibitory concentrations (MICs) ranging from 0.125 μg/mL to 1 μg/mL, while [Ru(phen)3]2+and [Ru(bpy)3]2+do not exhibit antimicrobial activity against the two bacterial genera studied at the concentration range used. When ciprofloxacin was combined with [Cr(phen)3]3+for the inhibition ofStaphylococcus aureusandEscherichia coli, an important synergistic effect was observed, FIC 0.066 forS. aureusand FIC 0.064 forE. coli. The work described here shows that chromium(III) complexes are bactericidal forS. aureusandE. coli. Our results indicate thatα-diimine chromium(III) complexes may be interesting to open new paths for metallodrug chemotherapy against different bacterial genera since some of these complexes have been found to exhibit remarkable antibacterial activities.


2021 ◽  
Vol 22 (15) ◽  
pp. 7959
Author(s):  
Federica Armas ◽  
Adriana Di Stasi ◽  
Mario Mardirossian ◽  
Antonello A. Romani ◽  
Monica Benincasa ◽  
...  

The emergence of multidrug-resistant bacteria is a worldwide health problem. Antimicrobial peptides have been recognized as potential alternatives to conventional antibiotics, but still require optimization. The proline-rich antimicrobial peptide Bac7(1-16) is active against only a limited number of Gram-negative bacteria. It kills bacteria by inhibiting protein synthesis after its internalization, which is mainly supported by the bacterial transporter SbmA. In this study, we tested two different lipidated forms of Bac7(1-16) with the aim of extending its activity against those bacterial species that lack SbmA. We linked a C12-alkyl chain or an ultrashort cationic lipopeptide Lp-I to the C-terminus of Bac7(1-16). Both the lipidated Bac-C12 and Bac-Lp-I forms acquired activity at low micromolar MIC values against several Gram-positive and Gram-negative bacteria. Moreover, unlike Bac7(1-16), Bac-C12, and Bac-Lp-I did not select resistant mutants in E. coli after 14 times of exposure to sub-MIC concentrations of the respective peptide. We demonstrated that the extended spectrum of activity and absence of de novo resistance are likely related to the acquired capability of the peptides to permeabilize cell membranes. These results indicate that C-terminal lipidation of a short proline-rich peptide profoundly alters its function and mode of action and provides useful insights into the design of novel broad-spectrum antibacterial agents.


2019 ◽  
Vol 10 ◽  
pp. 1864-1872
Author(s):  
Prof. Teodora P. Popova

The effect of ionized aqueous solutions (anolytes and catholyte) in the processing of fruits (cherries, morellos, and strawberries) for decontamination has been tested. Freshly prepared analytes and catholyte without the addition of salts were used, as well as stored for 7 months anolytes, prepared with 0.5% NaCl and a combination of 0.5% NaCl and 0.5% Na2CO3. The anolyte prepared with a combination of 0.5% NaCl and 0.5% Na2CO3, as well as the anolyte obtained with 0.5% NaCl, exhibit high antimicrobial activity against the surface microflora of strawberries, cherries, and sour cherries. They inactivate E. coli for 15 minutes. The other species of the fam. Enterobacteriaceae were also affected to the maximum extent, as is the total number of microorganisms, especially in cherries and sour cherries. Even stored for 7 months, they largely retain their antimicrobial properties. Anolyte and catholyte, obtained without the addition of salts, showed a lower effect on the total number of microorganisms, but had a significant effect on Gram-negative bacteria, and especially with regard to the sanitary indicative E. coli.


2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Tessa B. Moyer ◽  
Ashleigh L. Purvis ◽  
Andrew J. Wommack ◽  
Leslie M. Hicks

Abstract Background Plant defensins are a broadly distributed family of antimicrobial peptides which have been primarily studied for agriculturally relevant antifungal activity. Recent studies have probed defensins against Gram-negative bacteria revealing evidence for multiple mechanisms of action including membrane lysis and ribosomal inhibition. Herein, a truncated synthetic analog containing the γ-core motif of Amaranthus tricolor DEF2 (Atr-DEF2) reveals Gram-negative antibacterial activity and its mechanism of action is probed via proteomics, outer membrane permeability studies, and iron reduction/chelation assays. Results Atr-DEF2(G39-C54) demonstrated activity against two Gram-negative human bacterial pathogens, Escherichia coli and Klebsiella pneumoniae. Quantitative proteomics revealed changes in the E. coli proteome in response to treatment of sub-lethal concentrations of the truncated defensin, including bacterial outer membrane (OM) and iron acquisition/processing related proteins. Modification of OM charge is a common response of Gram-negative bacteria to membrane lytic antimicrobial peptides (AMPs) to reduce electrostatic interactions, and this mechanism of action was confirmed for Atr-DEF2(G39-C54) via an N-phenylnaphthalen-1-amine uptake assay. Additionally, in vitro assays confirmed the capacity of Atr-DEF2(G39-C54) to reduce Fe3+ and chelate Fe2+ at cell culture relevant concentrations, thus limiting the availability of essential enzymatic cofactors. Conclusions This study highlights the utility of plant defensin γ-core motif synthetic analogs for characterization of novel defensin activity. Proteomic changes in E. coli after treatment with Atr-DEF2(G39-C54) supported the hypothesis that membrane lysis is an important component of γ-core motif mediated antibacterial activity but also emphasized that other properties, such as metal sequestration, may contribute to a multifaceted mechanism of action.


2016 ◽  
Vol 60 (10) ◽  
pp. 5995-6002 ◽  
Author(s):  
Kristin R. Baker ◽  
Bimal Jana ◽  
Henrik Franzyk ◽  
Luca Guardabassi

ABSTRACTThe envelope of Gram-negative bacteria constitutes an impenetrable barrier to numerous classes of antimicrobials. This intrinsic resistance, coupled with acquired multidrug resistance, has drastically limited the treatment options against Gram-negative pathogens. The aim of the present study was to develop and validate an assay for identifying compounds that increase envelope permeability, thereby conferring antimicrobial susceptibility by weakening of the cell envelope barrier in Gram-negative bacteria. A high-throughput whole-cell screening platform was developed to measureEscherichia colienvelope permeability to a β-galactosidase chromogenic substrate. The signal produced by cytoplasmic β-galactosidase-dependent cleavage of the chromogenic substrate was used to determine the degree of envelope permeabilization. The assay was optimized by using known envelope-permeabilizing compounds andE. coligene deletion mutants with impaired envelope integrity. As a proof of concept, a compound library comprising 36 peptides and 45 peptidomimetics was screened, leading to identification of two peptides that substantially increased envelope permeability. Compound 79 reduced significantly (from 8- to 125-fold) the MICs of erythromycin, fusidic acid, novobiocin and rifampin and displayed synergy (fractional inhibitory concentration index, <0.2) with these antibiotics by checkerboard assays in two genetically distinctE. colistrains, including the high-risk multidrug-resistant, CTX-M-15-producing sequence type 131 clone. Notably, in the presence of 0.25 μM of this peptide, both strains were susceptible to rifampin according to the resistance breakpoints (R> 0.5 μg/ml) for Gram-positive bacterial pathogens. The high-throughput screening platform developed in this study can be applied to accelerate the discovery of antimicrobial helper drug candidates and targets that enhance the delivery of existing antibiotics by impairing envelope integrity in Gram-negative bacteria.


Author(s):  
Amita Shobha Rao ◽  
Shobha Kl ◽  
Prathibha Md’almeida ◽  
Kiranmai S Rai

  Objective: Infections caused by Gram-negative bacteria are important causes of morbidity and mortality. Extracts of plants and herbs such as Clitorea ternatea are used as diuretic. This work attempts to find out antimicrobial activity of aqueous and alcoholic extract of C. ternatea roots against Pseudomonas aeruginosa (ATCC 27853), Escherichia coli (ATCC 25922), clinical strains of Klebsiella pneumoniae, and Candida albicans.Methods: The agar well-diffusion method was done using Mueller Hinton agar and Sabouraud’s dextrose agar. The microorganism grown in peptone water was inoculated into culture medium. 4 mm diameter well punched into the agar was filled with 20 μl of aqueous and alcoholic root extracts C. ternatea extracts in various concentrations (100-25 μg/ml). The plates were incubated and antimicrobial activity was evaluated.Results: Aqueous root extract of C. ternatea with the concentration of 100 μg/ml showed zone of inhibition against E. coli (ATCC 25922) 18 mm, P. aeruginosa (ATCC 27853) 14 mm, multidrug resistant strain of K. pneumoniae 15 mm. Alcoholic extract of C. ternatea with the concentration of 100 μg/ml showed zone of inhibition of 35 mm against E. coli (ATCC 25922), P. aeruginosa (ATCC 27853) 22 mm, and multidrug resistant strain of K. pneumoniae 28 mm. C. albicanswas resistant to both extract of C. ternatea root. Conclusions: Alcoholic extract of C. ternatea is a better antibacterial agent against multidrug resistant Klebsiella species and other Gram-negative pathogens. Further, studies are required to identify active substances from the alcoholic extracts of C. ternatea for treating infections.


Author(s):  
M. Y. Iliyasu ◽  
I. Mustapha ◽  
H. Yakubu ◽  
H. M. Shuaibu ◽  
A. F. Umar ◽  
...  

Background of Study: Many virulence determinants contribute to the pathogenicity of Gram negative bacteria, like Escherichia coli, which is the most common cause of many infections worldwide such as urinary tract infection (UTI), profuse diarrhoea and septicaemia. Aim: To determine the genotypic characteristics of adhesin-producing E. coli isolates from clinical specimens. Place and Duration of Study: Conducted at the Infectious diseases hospital Bayara, Bauchi state, Nigeria, between February to March, 2019. Methods: A total of twelve (12) Gram negative bacterial isolates were selected based on the ability to grow on Luria-Bertani (LB) agar medium containing 100 µg/ml ampicillin. The isolates were from urine, stool, and blood specimens. The isolates were screened for multidrug resistant pattern according to Kirby-Bauer disc diffusion method. Adhesion factors, Fimbrial adhesin (fimH) and Invasive plasmid adhesin (ipaH) was genotyped by conventional PCR and sequenced. Results: All the isolates were resistant to Ampicillin, Cephalothin, Erythromycin, Fusidic acid, Novobiocin and Oxacillin, but sensitive to Augmentin, Colistin sulphate and Imipenem. Presence of fimH and ipaH genes were observed in nine isolates that expressed strong relationship with. Multidrug resistance (MDR). The fimH was the most prevalent found in urine, stool and blood isolates. Most of the adhesion genes sequence (61.8%) in this study had significant alignment (95 to 100% homology) with E.coli genome in the NCBI database. Conclusion: This study revealed the role of adhesin as virulence markers in MDR Gram negative bacteria and FimH is one of the commonest gene in MDR E.coli pathotypes.


2020 ◽  
Vol 18 (1) ◽  
pp. 72-78 ◽  
Author(s):  
Fatima Benyoucef ◽  
Mohammed El Amine Dib ◽  
Boufeldja Tabti ◽  
Arrar Zoheir ◽  
Jean Costa ◽  
...  

Background: Antibiotic resistance is today one of the most serious threats to global health, food security and development. Due to the growing number of infections, treatment becomes more difficult, if not impossible, because of the loss of antibiotic efficacy. Objective: In the present investigation, the chemical composition of essential oils of Ammoides verticillata and Satureja candidissima and their synergistic effects on antimicrobial activities were investigated. Methods: The chemical composition of the essential oil was analyzed by Gas Chromatography (GC) and Gas Chromatography-Mass Spectroscopy (GC/MS). The antimicrobial activity of the essential oils was investigated using dilution-agar method against nine bacterial strains three Gram-negative Escherichia coli (ATCC 25922), Pseudomonas aeruginosa (ATCC 27853) and Salmonella typhi (ATCC 14028), and six Gram-positive: Staphylococcus aureus (ATCC 43300,) Clostridium sporogenes (ATCC 19404), Bacillus subtilis (ATCC 6633), Enterococcus faecalis (ATCC 7314), Lactobacillus rhamnosus (ATCC 53103) and Bacillus cereus (ATCC 14579). Results: The essential oil of A. verticillata was characterized principally by carvacrol (44,3%), Limonene (19,3%) and p-cymene (19,2%). The constituents identified of S. candidissima essential oil were principally oxygenated monoterpenes represented by pulegone (70,4%). The essential oil of A. verticillata had a good antimicrobial activity against four bacterial strains (Escherichia coli, Salmonella typhi, Lactobacillus rhamnosus and Bacillus cereus) with MIC and MBC values between 0.2-0.4 µl/ml and 0.2-6.2 µl/ml, respectively. While, S. candidissima essential oil had moderate antimicrobial activities against all strains with MIC and MBC values between 1.5-6.2 µl/ml and 6.2-12.5 µl/ml, respectively. The results of antimicrobial activity of essential oils blend presented higher antimicrobial activity against all tested bacteria with MIC and MBC values between 0.3-1.5 µl/ml and 0.4-6.2 µl/ml, respectively. Conclusion: The essential oils blend presented high antimicrobial activity compared to virgin oils. This activity can be due to the association of active compounds such as carvacrol and pulegone. These findings provide a new source of drugs that may help in therapy to lead to the development of a new treatment based on a combination of these essential oils against gram-negative and gram-positive bacteria that continue to pose a threat to public health.


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