Antimicrobial efficacy of Cecropin A (1–7)- Melittin and Lactoferricin (17–30) against multi-drug resistant Salmonella Enteritidis

Gourkhede, Diksha; Bhoomika, Sirsant; Pathak, Richa; Yadav, Jay Prakash; Nishanth, Dani; Vergis, Jess; Malik, S.V.S.; Barbuddhe, Sukhadeo B.; Rawool, Deepak B.

doi:10.1016/j.micpath.2020.104405

Cited by 8 publications

(18 citation statements)

References 44 publications

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“…The haemolytic assay of the individual phytochemical was performed by measuring the release of haemoglobin from sheep and human ‘O’ erythrocytes at 540 nm [ 44 ]. The percentage of haemolysis was calculated as (A sample – A PBS )/ (A Triton−X – A PBS ) ×100, in which A sample is the absorbance of phytochemical treatment, A PBS is the absorbance of untreated control with PBS, and A Triton−X is the absorbance of lysed cells treated with Triton X-100 measured at 540 nm.…”

Section: Methodsmentioning

confidence: 99%

In silico molecular docking and in vitro antimicrobial efficacy of phytochemicals against multi-drug-resistant enteroaggregative Escherichia coli and non-typhoidal Salmonella spp.

et al. 2021

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Background In the wake of emergence of antimicrobial resistance, bioactive phytochemical compounds are proving to be important therapeutic agents. The present study envisaged in silico molecular docking as well as in vitro antimicrobial efficacy screening of identified phytochemical ligands to the dispersin (aap) and outer membrane osmoporin (OmpC) domains of enteroaggregative Escherichia coli (EAEC) and non-typhoidal Salmonella spp. (NTS), respectively. Materials and methods The evaluation of drug-likeness, molecular properties, and bioactivity of the identified phytocompounds (thymol, carvacrol, and cinnamaldehyde) was carried out using Swiss ADME, while Protox-II and StopTox servers were used to identify its toxicity. The in silico molecular docking of the phytochemical ligands with the protein motifs of dispersin (PDB ID: 2jvu) and outer membrane osmoporin (PDB ID: 3uu2) were carried out using AutoDock v.4.20. Further, the antimicrobial efficacy of these compounds against multi-drug resistant EAEC and NTS strains was determined by estimating the minimum inhibitory concentrations and minimum bactericidal concentrations. Subsequently, these phytochemicals were subjected to their safety (sheep and human erythrocytic haemolysis) as well as stability (cationic salts, and pH) assays. Results All the three identified phytochemicals ligands were found to be zero violators of Lipinski’s rule of five and exhibited drug-likeness. The compounds tested were categorized as toxicity class-4 by Protox-II and were found to be non- cardiotoxic by StopTox. The docking studies employing 3D model of dispersin and ompC motifs with the identified phytochemical ligands exhibited good binding affinity. The identified phytochemical compounds were observed to be comparatively stable at different conditions (cationic salts, and pH); however, a concentration-dependent increase in the haemolytic assay was observed against sheep as well as human erythrocytes. Conclusions In silico molecular docking studies provided useful insights to understand the interaction of phytochemical ligands with protein motifs of pathogen and should be used routinely before the wet screening of any phytochemicals for their antibacterial, stability, and safety aspects.

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Section: Methodsmentioning

confidence: 99%

In silico molecular docking and in vitro antimicrobial efficacy of phytochemicals against multi-drug-resistant enteroaggregative Escherichia coli and non-typhoidal Salmonella spp.

et al. 2021

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“…Although the tested AMPs were found potentially effective against the pathogens outside the eukaryotic cells and were not much effective against intracellular pathogens like Salmonella spp. (Gourkhede et al, 2020), CPPs were suggested as better alternatives for intracellular pathogens. Nevertheless, a few studies were carried out to explore the antibacterial efficacy of different CPPs against intracellular sensitive strains of NTS, particularly S. Typhimurium (Zhu and Shin, 2009;Li et al, 2012;Li et al, 2013).…”

Section: Selection and Designing Of Cama-cppmentioning

confidence: 99%

“…The CPP variant of CAMA was designed using online CellPPD software (https://webs.iiitd.edu.in/raghava/cellppd/ submission.php). Also, two known CPPs, viz., P7 and APP, suggested effective against S. Typhimurium (Li et al, 2012;Li et al, 2013), and the non-CPP version of CAMA (Gourkhede et al, 2020) was also evaluated in this study. All the peptides (Supplementary Table 1) commercially synthesized (Shanghai Science Biological Technology, China) were resuspended in sterile phosphate-buffered saline (PBS) and stored at À20 C until further use.…”

Section: Cell-penetrating Ampsmentioning

confidence: 99%

“…To investigate the stability of CPP-AMPs at physiological concentration of cationic salts (150 mM NaCl and 2 mM MgCl 2 ), corresponding peptides were co-incubated in the presence of 150 mM NaCl and 2 mM MgCl 2 , separately with each MDR strains of S. Enteritidis (n = 3) and S. Typhimurium (n = 3) in CA-MH broth (Gourkhede et al, 2020) and the antimicrobial efficacy was determined as described earlier.…”

Section: Effect Of Physiological Concentration Of Cationic Saltsmentioning

confidence: 99%

“…Nevertheless, CAMA was not found effective against intracellular pathogens such as Salmonella spp. (Gourkhede et al, 2020). Hence, in this study, we modified CAMA as a CPP variant (CAMA-CPP) and compared its antimicrobial efficacy with those CPPs that were suggested as effective against intracellular sensitive strains of S. Typhimurium (Li et al, 2012;Li et al, 2013).…”

Section: Introductionmentioning

confidence: 99%

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Antibacterial efficacy of in‐house designed cell‐penetrating peptide against multi‐drug resistant strains of Salmonella Enteritidis and Salmonella Typhimurium

Nishanth

Bhoomika

Gourkhede

et al. 2021

Environmental Microbiology

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Summary The in vitro antibacterial efficacy of an in‐house designed cell‐penetrating peptide (CPP) variant of Cecropin A (1–7)‐Melittin (CAMA) (CAMA‐CPP) against the characterized multi‐drug resistant (MDR) field strains of Salmonella Enteritidis and Salmonella Typhimurium were evaluated and compared with two identified CPPs namely, P7 and APP, keeping CAMA as control. Initially, the minimum inhibitory concentration (MIC) (μg ml−1) of in‐house designed CAMA‐CPP, APP and CAMA was determined to be 3.91, whereas that of P7 was 7.81; however, the minimum bactericidal concentration (MBC) of all the peptides were twice the MIC. CAMA‐CPP and CAMA were found to be stable under different conditions (high‐end temperatures, proteinase‐K, cationic salts, pH and serum) when compared to the other CPPs. Moreover, CAMA‐CPP exhibited negligible cytotoxicity in HEp‐2 and RAW 264.7 cell lines as well as haemolysis in the sheep and human erythrocytes with no adverse effects against the commensal gut lactobacilli. In vitro time‐kill assay revealed that the MBC levels of CAMA‐CPP and APP could eliminate the intracellular MDR‐Salmonella infections from mammalian cell lines; however, CAMA and P7 peptides were ineffective. CAMA‐CPP appears to be a promising antimicrobial candidate and opens up further avenues for its in vivo clinical translation.

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Therapeutic Potential of CPPs

Langel

2023

CPP, Cell-Penetrating Peptides

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Antimicrobial efficacy of Cecropin A (1–7)- Melittin and Lactoferricin (17–30) against multi-drug resistant Salmonella Enteritidis

Cited by 8 publications

References 44 publications

In silico molecular docking and in vitro antimicrobial efficacy of phytochemicals against multi-drug-resistant enteroaggregative Escherichia coli and non-typhoidal Salmonella spp.

In silico molecular docking and in vitro antimicrobial efficacy of phytochemicals against multi-drug-resistant enteroaggregative Escherichia coli and non-typhoidal Salmonella spp.

Antibacterial efficacy of in‐house designed cell‐penetrating peptide against multi‐drug resistant strains of Salmonella Enteritidis and Salmonella Typhimurium

Therapeutic Potential of CPPs

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