Recombinant Production and Antimicrobial Assessment of Beta Casein- IbAMP4 as a Novel Antimicrobial Polymeric Protein and its Synergistic Effects with Thymol

Fahimirad, Shohreh; Razavi, Seyed Hadi; Abtahi, Hamid; Alizadeh, Houshang; Ghorbanpour, Mansour

doi:10.1007/s10989-017-9605-5

Cited by 21 publications

(3 citation statements)

References 40 publications

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“…Additionally, the broad utilization of chemical disinfectants has caused an expanded growing resistance to multiple biocidal agents and pathogenic resistance ( Fahimirad et al, 2017 ; Fahimirad et al, 2018 ; Nadi et al, 2020 ). Thus, there is an urgent need for novel methods in removing bacteria from water supplies which meet the requirement of effective antimicrobial activity, superior filtration flux with acceptable retention potentials ( Zodrow et al, 2014 ).…”

Section: Introductionmentioning

confidence: 99%

Efficient removal of water bacteria and viruses using electrospun nanofibers

Fahimirad

Sillanpää

2021

Science of The Total Environment

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130

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Pathogenic contamination has been considered as a significant worldwide water quality concern. Due to providing promising opportunities for the production of nanocomposite membranes with tailored porosity, adjustable pore size, and scaled-up ability of biomolecules incorporation, electrospinning has become the center of attention. This review intends to provide a detailed summary of the recent advances in the fabrication of antibacterial and antiviral electrospun nanofibers and discuss their application efficiency as a water filtration membrane. The current review attempts to give a functionalist perspective of the fundamental progress in construction strategies of antibacterial and antiviral electrospun nanofibers. The review provides a list of antibacterial and antiviral agents commonly used as water membrane filters and discusses the challenges in the incorporation process. We have thoroughly studied the recent application of functionalized electrospun nanofibers in the water disinfection process, with an emphasis on their efficiency. Moreover, different antibacterial and antiviral assay techniques for membranes are discussed, the gaps and limitations are highlighted and promising strategies to overcome barriers are studies.

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Section: Introductionmentioning

confidence: 99%

Efficient removal of water bacteria and viruses using electrospun nanofibers

Fahimirad

Sillanpää

2021

Science of The Total Environment

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130

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“…values were poured into a microtube and incubated at 16 °C overnight [9] . Expression, purification and refolding optimization: Synthesized plasmids were transferred to E. coli BL21 (DE3) competent cells according to the Standard thermal shock protocol [10] . Finally, in three conditions, 50µl and 100µl, and bacterial deposits were cultured in nutrient agar (NA) containing ampicillin (100 mg/mL) and incubated at °C for 24h.…”

Section: Gene Constructionmentioning

confidence: 99%

LL37-rIb-AMP4 Hybrid Peptide as a Therapy for Systematic Infections of Acinetobacter baumannii, Pseudomonas aeruginosa, Vancomycin-resistant Enterococcus (VRE), and Methicillin-resistant Staphylococcus aureus (MRSA) cells in a mouse model.

Sadoogh Abbasian,

Ghaznavi-Rad

et al. 2023

IEM

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Background: Antimicrobial peptides (AMPs) are beneficial compounds that could be used as a new and effective method to suppress microbes. Both Ib-AMP4 and LL37 are antimicrobial peptides with a wide range of antimicrobial activities. This research aimed to evaluate the antibacterial potential of LL37-rIb-AMP4 hybrid protein as an antimicrobial agent against pathogenic bacteria. Therefore, its antibacterial effects against Acinetobacter baumannii, Pseudomonas aeruginosa, vancomycin-resistant Enterococcus (VRE), and methicillinresistant Staphylococcus aureus (MRSA) were investigated in vivo and in vitro.Materials & Methods: In this study, antimicrobial peptides rIb-AMP4, LL37, and LL37-rIb-AMP4 were expressed, purified, and refolded, and their synergistic and antibacterial effects in combination with each other (LL37+rIb-AMP4) and as fusion proteins (LL37-rIb-AMP4) were tested against A. baumannii, P. aeruginosa, VRE, and MRSA cells in vitro (MIC, time kill, and SEM) and against P. aeruginosa and VRE cells in vivo. Findings: LL37-rIb-AMP4 Protein with molecular weight= 28 KD was correctly produced and purified. Despite the lack of synergistic effects between LL37 and rIb-AMP4 peptides in vitro, the stability test results showed higher stability for LL37-rIb-AMP4 hybrid protein.The findings of in vivo tests confirmed that all infected mice were improved with LL37-rIb-AMP4 and no signs of bacteria were observed in their blood and spleen samples. Also, these results confirmed the stability and higher activity of LL37-rIb-AMP4 than the single form of these proteins. Conclusion:Considering the antimicrobial potential of the produced proteins, it seems that the recombinant LL37-rIb-AMP4 protein could be considered and used as a stable and active antimicrobial drug in future studies.

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“…After 24 hours of incubation at 37°C, 20 µL of resazurin dyes (0.02% w/v) was added to each well, and the plates were incubated for another 2 hours. The concentration of each treatment group in the last blue-colored well was deemed to be the MIC value (30).…”

Section: Antimicrobial Activity Assays Of Ll-37 and Oncorhyncin IImentioning

confidence: 99%

Evaluation of the Synergistic Effect of LL-37 and Oncorhyncin II Recombinant Proteins on Staphylococcus aureus Under In Vitro Conditions

Safari

Tabatabaei

Abtahi

et al. 2023

Jundishapur J Microbiol

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Background: The treatment of Staphylococcus aureus infections has become a public health crisis due to the extensive development of antimicrobial resistance. Antimicrobial peptides (AMPs) have been introduced as promising naturally-derived antimicrobial alternatives to antibiotics. LL-37 and oncorhyncin II are 2 AMPs with notable proven antibacterial effects. Objectives: This study aimed to produce recombinant LL-37 and oncorhyncin II and investigate their synergistic effects on S. aureus (ATCC25923). Methods: The synthetic genes of LL-37 and oncorhyncin II were individually ligated into the pET32a expression vector. Transformed pET32a was introduced into Escherichia coli BL21 as an expression host. The protein expression and purification steps were optimized, and the biological effectiveness of the peptides was evaluated by assessing the minimum inhibitory concentration (MIC), time-kill, and growth kinetic tests against S. aureus. Results: The MIC assay confirmed the effective antibacterial performances of LL-37 and oncorhyncin II against S. aureus at 30.6 and 47.93 µg/mL, respectively. The peptides’ synergistic activity was validated by the checkerboard method. A combination of LL-37 and oncorhyncin II at 2 × MIC showed a sharp decline of the viable cells with over 3-time reductions in log 10 colony-forming units (CFU)/mL within the first 5 hours. The growth kinetic results confirmed the high effectiveness of the peptides’ combination in eliminating the bacterial inoculum turbidity by 50% reduction during the first hour of exposure. Conclusions: The produced recombinant LL-37 and oncorhyncin II showed effective antimicrobial function against S. aureus. The synergistic performance of the peptides was repeatedly confirmed through checkerboard, time-kill, and growth kinetic assays.

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Recombinant Production and Antimicrobial Assessment of Beta Casein- IbAMP4 as a Novel Antimicrobial Polymeric Protein and its Synergistic Effects with Thymol

Cited by 21 publications

References 40 publications

Efficient removal of water bacteria and viruses using electrospun nanofibers

Efficient removal of water bacteria and viruses using electrospun nanofibers

LL37-rIb-AMP4 Hybrid Peptide as a Therapy for Systematic Infections of Acinetobacter baumannii, Pseudomonas aeruginosa, Vancomycin-resistant Enterococcus (VRE), and Methicillin-resistant Staphylococcus aureus (MRSA) cells in a mouse model.

Evaluation of the Synergistic Effect of LL-37 and Oncorhyncin II Recombinant Proteins on Staphylococcus aureus Under In Vitro Conditions

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