Antimicrobial Moieties Research Articles

Efficacious Extraction and Purification Technique of a Potential Antimycobacterial Bacteriocin Produced by Bacillus subtilis (MK733983) of Ethnomedicinal Origin.

The rapid proliferation of antibiotic-resistant microbes has imposed an urgent need for development of novel antimicrobial agents with diverse mechanisms. This study reports a novel extraction method with salting-in and salting-out method for obtaining potential bacteriocin from Bacillus subtilis (MK733983) of ethnomedicinal origin. This technique extracted bacteriocin with desired antimicrobial peptide moieties that showed creditable minimum inhibitory concentrations, thermostability and efficacy compared to all other extraction protocols attempted. Further study used a unique scheme of steps in RP-HPLC purification process using methanol-water as solvents for the bacteriocin that achieved an outstanding antimicrobial activity against Staphylococcus aureus (MTCC 737). The bacteriocin is sensitive to proteases, confirming its proteinaceous nature and showed promising heat stability up to 70°C for 10min. Bacteriocin extracted from a series of ammonium sulphate precipitation showed MIC values 350µg and 300µg for Mycobacterium smegmatis and Staphylococcus aureus respectively. On the other hand, bacteriocin extracted by using chloroform showed MIC values 400µg and 300µg for M. smegmatis and Staphylococcus aureus. All the results implicate the efficacy of bacteriocin and future prospect as an effective antimicrobial agent.

Incorporating Antimicrobial Activity During Synthesis of New Acid-Azo Dyes: Thermal Stability and Application on Various Fabrics.

Textile materials are susceptible to microbial attack as they provide suitable conditions for their growth. The microbes grow with normal body secretions on garments. These microbes are responsible for the weakening, brittleness, and discoloration of the substrate. Furthermore, they cause many health issues to the wearer, including dermal infection, bad odour etc. They threaten the human health as well as create tenderness in fabric. Usually, antimicrobial textiles are prepared by applying antimicrobial finishes after dyeing, which is an expensive approach. Concerning these adversities, in the present study, a series of antimicrobial acid-azo dyes have been synthesized by incorporating antimicrobial sulphonamide moiety into the dye molecules during its synthesis. A commercially available sulphonamide-based compound, sulfadimidine Na-salt was used as a diazonium component and coupled with different aromatic amines to get desired dye molecules. Since dyeing and finishing are two separate energy-intensive processes, in the current research work, an approach to combine both processes in one step has been adopted that would be economical, time-saving, and environment friendly. Structures of the resultant dye molecules have been confirmed using different spectral techniques such as Mass spectrometry, 1H-NMR spectroscopy, FT-IR, and UV-Visible spectroscopy. Thermal stability of the synthesized dyes was also determined. These dyes have been applied to wool and nylon-6 fabrics. Their various fastness properties were examined using ISO standard methods. All the compounds exhibited good to excellent fastness properties. The synthesized dyes and the dyed fabrics were screened biologically against Staphylococcus aureus ATCC 6538 and Escherichia coli ATCC 10536, resulting in significant antibacterial activities.

Supramolecular Self-Healing Antifouling Coating for Dental Materials.

In orthodontic treatment, orthodontic appliances are prone to bacterial infections, which pose a risk to oral health. Surface modification of orthodontic appliances has been explored to improve their antifouling properties and impart antibacterial capabilities, inhibiting initial bacterial adhesion and biofilm formation. However, coatings are susceptible to damage in the complex oral environment, leading to a loss of functionality. Here, we have prepared an antifouling self-healing coating based on supramolecular bonding by employing a simple spin coating method. The presence of the hydrophilic zwitterionic trimethylamine N-oxide (TMAO) and the hydrophobic antimicrobial moieties triclosan acrylate (TCSA) imparts to the polymers an amphiphilic structure and enhances the interaction with bacteria, resulting in excellent antimicrobial activity and surface antifouling properties. The multiple hydrogen bonds of ureido-pyrimidinone methacrylate (UPyMA) and ionic interactions contained in the polymers not only increased the adhesion of the coating to the material substrate (approximately 3 times) but also endowed the coating with the intrinsic self-healing ability to restore the antibiofouling properties at oral temperature and humidity. Finally, the polymer coating is biologically safe both in vitro and in vivo, showing no cytotoxic effects on cells and tissues. This research offers a promising avenue for improving the performance of orthodontic appliances and contributes to the maintenance and treatment of oral health.

Designing New Hybrid Antibiotics: Proline-Rich Antimicrobial Peptides Conjugated to the Aminoglycoside Tobramycin.

Resistance to aminoglycoside antibiotics is a serious problem, typically arising from inactivating enzymes, reduced uptake, or increased efflux in the important pathogens for which they are used as treatment. Conjugating aminoglycosides to proline-rich antimicrobial peptides (PrAMPs), which also target ribosomes and have a distinct bacterial uptake mechanism, might mutually benefit their individual activities. To this aim we have developed a strategy for noninvasively modifying tobramycin to link it to a Cys residue and through this covalently link it to a Cys-modified PrAMP by formation of a disulfide bond. Reduction of this bridge in the bacterial cytosol should release the individual antimicrobial moieties. We found that the conjugation of tobramycin to the well-characterized N-terminal PrAMP fragment Bac7(1-35) resulted in a potent antimicrobial capable of inactivating not only tobramycin-resistant bacterial strains but also those less susceptible to the PrAMP. To a certain extent, this activity also extends to the shorter and otherwise poorly active fragment Bac7(1-15). Although the mechanism that allows the conjugate to act when its individual components do not is as yet unclear, results are very promising and suggest this may be a way of resensitizing pathogens that have developed resistance to the antibiotic.

Antimicrobial nano-assemblies of tryptocidine C, a tryptophan-rich cyclic decapeptide, from ethanolic solutions

Tryptocidine C (TpcC), a Trp-rich cyclodecapeptide is a minor constituent in the antibiotic tyrothricin complex from Brevibacillus parabrevis. TpcC possesses a high tendency to oligomerise in aqueous solutions and dried TpcC forms distinct self-assembled nanoparticles. High-resolution scanning electron microscopy revealed the influence of different ethanol:water solvent systems on TpcC self-assembly, with the TpcC, dried from a high concentration in 15% ethanol, primarily assembling into small nanospheres with 24.3 nm diameter and 0.05 polydispersity. TpcC at 16 μM, near its CMC, formed a variety of structures such as small nanospheres, large dense nanospheroids and facetted 3-D-crystals, as well as sheets and coarse carpet-like structures which depended on ethanol concentration. Drying 16 μM TpcC from 75% ethanol resulted in highly facetted 3-D crystals, as well as small nanospheres, while those in 10% ethanol preparation had less defined facets. Drying from 20 to 50% ethanol led to polymorphic architectures with a few defined nanospheroids and various small nanoparticles, imbedded in carpet- and sheet-like structures. These polymorphic surface morphologies correlated with maintenance of fluorescence properties and the surface-derived antibacterial activity against Staphylococcus aureus over time, while there was a significant change in fluorescence and loss in activity in the 10% and 75% preparations where 3-D crystals were observed. This indicated that TpcC oligomerisation in solutions with 20–50% ethanol leads to metastable structures with a high propensity for release of antimicrobial moieties, while those leading to crystallisation limit active moieties release. TpcC nano-assemblies can find application in antimicrobial coatings, surface disinfectants, food packaging and wound healing materials.

Molecular Entities of Antimicrobial Drugsand Resistance Mechanism Underlying: Bioaccessibility, Bioavailability, and Bioaccumulation

Molecular Entities of Antimicrobial Drugsand Resistance Mechanism Underlying: Bioaccessibility, Bioavailability, and Bioaccumulation

Design and in silico analysis of a whole-cell biosensor able to kill methicillin-resistant Staphylococcus aureus.

The rise of methicillin-resistant Staphylococcus aureus (MRSA) infections has gained concern throughout the world over the past decades. Alternative therapeutic agents to antibiotics are rapidly growing to impede the proliferation of MRSA-caused infections. Lately, synthetic biology techniques have developed whole-cell biosensors by designing gene circuitry capable of sensing quorum-sensing (QS) molecules of pathogens and triggering expression of an antimicrobial moiety that kills MRSA and therefore prevents its further proliferation. Here, an E. coli was engineered in silico to act as a whole-cell biosensor that senses QS molecules from MRSA and triggers the expression of a bacteriocin that kills MRSA. To achieve this functionality, biosensor and bacteriocin modules were constructed and assembled into a vector. Both modules were codon-optimized to increase the yield production of the recombinant proteins. We then demonstrate in silico that the construction of a dual biosensor-killer plasmid, which holds two genetical modules known as biosensor and bacteriocin modules, enables the recombinant host to sense QS molecules from MRSA. Our designed whole-cell biosensor demonstrates in silico its ability to produce and secrete the bacteriocin as a function of the external concentration of autoinducer peptide from MRSA. These in silico results unravel the possibility of designing antimicrobial smarter therapeutics against resistant pathogens.

Conjugation of Synthetic Polyproline Moietes to Lipid II Binding Fragments of Nisin Yields Active and Stable Antimicrobials.

Coupling functional moieties to lantibiotics offers exciting opportunities to produce novel derivatives with desirable properties enabling new functions and applications. Here, five different synthetic hydrophobic polyproline peptides were conjugated to either nisin AB (the first two rings of nisin) or nisin ABC (the first three rings of nisin) by using click chemistry. The antimicrobial activity of nisin ABC + O6K3 against Enterococcus faecium decreased 8-fold compared to full-length nisin, but its activity was 16-fold better than nisin ABC, suggesting that modifying nisin ABC is a promising strategy to generate semi-synthetic nisin hybrids. In addition, the resulting nisin hybrids are not prone to degradation at the C-terminus, which has been observed for nisin as it can be degraded by nisinase or other proteolytic enzymes. This methodology allows for getting more insight into the possibility of creating semi-synthetic nisin hybrids that maintain antimicrobial activity, in particular when synthetic and non-proteinaceous moieties are used. The success of this approach in creating viable nisin hybrids encourages further exploring the use of different modules, e.g., glycans, lipids, active peptide moieties, and other antimicrobial moieties.

Isolation and Characterization of Lactobacillus spp. from Kefir Samples in Malaysia.

Kefir is a homemade, natural fermented product comprised of a probiotic bacteria and yeast complex. Kefir consumption has been associated with many advantageous properties to general health, including as an antioxidative, anti-obesity, anti-inflammatory, anti-microbial, and anti-tumor moiety. This beverage is commonly found and consumed by people in the United States of America, China, France, Brazil, and Japan. Recently, the consumption of kefir has been popularized in other countries including Malaysia. The microflora in kefir from different countries differs due to variations in culture conditions and the starter media. Thus, this study was aimed at isolating and characterizing the lactic acid bacteria that are predominant in Malaysian kefir grains via macroscopic examination and 16S ribosomal RNA gene sequencing. The results revealed that the Malaysian kefir grains are dominated by three different strains of Lactobacillus strains, which are Lactobacillus harbinensis, Lactobacillusparacasei, and Lactobacillus plantarum. The probiotic properties of these strains, such as acid and bile salt tolerances, adherence ability to the intestinal mucosa, antibiotic resistance, and hemolytic test, were subsequently conducted and extensively studied. The isolated Lactobacillus spp. from kefir H maintained its survival rate within 3 h of incubation at pH 3 and pH 4 at 98.0 ± 3.3% and 96.1 ± 1.7% of bacteria growth and exhibited the highest survival at bile salt condition at 0.3% and 0.5%. The same isolate also showed high adherence ability to intestinal cells at 96.3 ± 0.01%, has antibiotic resistance towards ampicillin, penicillin, and tetracycline, and showed no hemolytic activity. In addition, the results of antioxidant activity tests demonstrated that isolated Lactobacillus spp. from kefir G possessed high antioxidant activities for total phenolic content (TPC), total flavonoid content (TFC), ferric reducing ability of plasma (FRAP), and 1,1-diphenyl-2-picryl-hydrazine (DPPH) assay compared to other isolates. From these data, all Lactobacillus spp. isolated from Malaysian kefir serve as promising candidates for probiotics foods and beverage since they exhibit potential probiotic properties and antioxidant activities.

Surface-Attached Poly(oxanorbornene) Hydrogels with Antimicrobial and Protein-Repellent Moieties: The Quest for Simultaneous Dual Activity.

By copolymerizing an amphiphilic oxanorbornene monomer bearing N- tert-butyloxycarbonyl (Boc) protected cationic groups with an oxanorbornene-functionalized poly(ethylene glycol) (PEG) macromonomer, bifunctional comb copolymers were obtained. Varying the comonomer ratios led to copolymers with PEG contents between 5–25 mol %. These polymers were simultaneously surface-immobilized on benzophenone-bearing substrates and cross-linked with pentaerythritoltetrakis(3-mercapto­propionate). They were then immersed into HCl to remove the Boc groups. The thus obtained surface-attached polymer hydrogels (called SMAMP*-co-PEG) were simultaneously antimicrobial and protein-repellent. Physical characterization data showed that the substrates used were homogeneously covered with the SMAMP*-co-PEG polymer, and that the PEG moieties tended to segregate to the polymer–air interface. Thus, with increasing PEG content, the interface became increasingly hydrophilic and protein-repellent, as demonstrated by a protein adhesion assay. With 25 mol % PEG, near-quantitative protein-adhesion was observed. The antimicrobial activity of the SMAMP*-co-PEG polymers originates from the electrostatic interaction of the cationic groups with the negatively charged cell envelope of the bacteria. However, the SMAMP*-co-PEG surfaces were only fully active against E. coli, while their activity against S. aureus was already compromised by as little as 5 mol % (18.8 mass %) PEG. The long PEG chains seem to prevent the close interaction of bacteria with the surface, and also might reduce the surface charge density.

Postmarketing experience with Neutrolin\xae (taurolidine, heparin, calcium citrate) catheter lock solution in hemodialysis patients

Catheter-related bloodstream infections (CRBSI) are major complications for patients with life-threatening conditions requiring chronic vascular catheterization. The wide range of etiologic microbes and the ongoing development of resistance to antimicrobials with specific mechanisms of action make this an appropriate target for applying a nonspecific antimicrobial therapeutic. Taurolidine hydrolyzes into two antimicrobial moieties, formaldehyde and methylene glycol, which react with microbial surfaces. Neutrolin® (taurolidine, heparin, calcium citrate) was recently introduced in Germany as an antimicrobial catheter lock solution. This postmarketing experience collected data on 201 patients at 20 centers from January 2014 through September 2016. Likely CRBSI was observed in 13 episodes in 47,118 days (0.2759 per 1000 days [0.1468, 0.4718]). Thrombosed catheter was observed in seven catheters in 47,118 days (0.1486 per 1000 days [0.0595, 0.3061]). No adverse drug reactions that led to the discontinuation of Neutrolin® use were reported. Two patients experienced occasional transient dysgeusia. Neutrolin®, when used in conjunction with guideline-based catheter care, showed reduction in the rate of both CRBSI and catheter thrombosis relative to recent historical controls.

Polymeric Antimicrobial N-Halamine-Surface Modification of Stainless Steel

The surfaces of materials fabricated from stainless steel are challenging to functionalize with antimicrobial moieties. This work demonstrates that stainless steel surfaces can be modified with an N-halamine-based copolymer in order to obtain antimicrobial activity. In this regard, a copolymer (HACM) of 2-acrylamido-2-methyl-1-(5-methylhydantoinyl)propane and 3-cloro-2-hydroxypropyl methacrylate was synthesized and grafted onto a stainless steel surface via covalent attachment. Synthesized monomers and copolymers were characterized by NMR, FTIR, and XPS spectral analyses. Upon treatment with dilute bleach, the stainless steel surfaces were rendered antimicrobial, possessing a sufficient amount of chlorine content and excellent stability and durability. The modified stainless steel samples inactivated 6 logs of Staphylococcus aureus and Escherichia coli O157:H7 bacteria within 15 min of contact time. Stabilities of the coatings toward washing and UVA exposure were also studied. The stainless steel samples ...

Polymer-Based Surfaces Designed to Reduce Biofilm Formation: From Antimicrobial Polymers to Strategies for Long-Term Applications.

Contact-active antimicrobial polymer surfaces bear cationic charges and kill or deactivate bacteria by interaction with the negatively charged parts of their cell envelope (lipopolysaccharides, peptidoglycan, and membrane lipids). The exact mechanism of this interaction is still under debate. While cationic antimicrobial polymer surfaces can be very useful for short-term applications, they lose their activity once they are contaminated by a sufficiently thick layer of adhering biomolecules or bacterial cell debris. This layer shields incoming bacteria from the antimicrobially active cationic surface moieties. Besides discussing antimicrobial surfaces, this feature article focuses on recent strategies that were developed to overcome the contamination problem. This includes bifunctional materials with simultaneously presented antimicrobial and protein-repellent moieties; polymer surfaces that can be switched from an antimicrobial, cell-attractive to a cell-repellent state; polymer surfaces that can be regenerated by enzyme action; degradable antimicrobial polymers; and antimicrobial polymer surfaces with removable top layers.

Antimicrobial resistance challenged with metal-based antimicrobial macromolecules.

Antimicrobial resistance threatens the achievements of science and medicine, as it deactivates conventional antimicrobial therapeutics. Scientists respond to the threat by developing new antimicrobial platforms to prevent and treat infections from these resistant strains. Metal-based antimicrobial macromolecules are emerging as an alternative to conventional platforms because they combine multiple mechanisms of action into one platform due to the distinctive properties of metals. For example, metals interact with intracellular proteins and enzymes, and catalyse various intracellular processes. The macromolecular architecture offers a means to enhance antimicrobial activity since several antimicrobial moieties can be conjugated to the scaffold. Further, these macromolecules can be fabricated into antimicrobial materials for contact-killing medical implants, fabrics, and devices. As volatilization or leaching out of the antimicrobial moieties from the macromolecular scaffold is reduced, these medical implants, fabrics, and devices can retain their antimicrobial activity over an extended period. Recent advances demonstrate the potential of metal-based antimicrobial macromolecules as effective platforms that prevent and treat infections from resistant strains. In this review these advances are thoroughly discussed within the context of examples of metal-based antimicrobial macromolecules, their mechanisms of action and biocompatibility.

Mechanically Robust and Transparent N-Halamine Grafted PVA-co-PE Films with Renewable Antimicrobial Activity.

Antimicrobial polymeric films that are both mechanically robust and function renewable would have broad technological implications for areas ranging from medical safety and bioengineering to foods industry; however, creating such materials has proven extremely challenging. Here, a novel strategy is reported to create high-strength N-halamine incorporated poly(vinyl alcohol-co-ethylene) films (HAF films) with renewable antimicrobial activity by combining melt radical graft polymerization and reactive extrusion technique. The approach allows here the intrinsically rechargeable N-halamine moieties to be covalently incorporated into polymeric films with high biocidal activity and durability. The resulting HAF films exhibit integrated properties of robust mechanical strength, high transparency, rechargeable chlorination capability (>300 ppm), and long-term durability, which can effectively offer 3-5 logs CFU reduction against typical pathogenic bacterium Escherichia coli within a short contact time of 1 h, even at high organism conditions. The successful synthesis of HAF films also provides a versatile platform for exploring the applications of antimicrobial N-halamine moieties in a self-supporting, structurally adaptive, and function renewable form.

Development of a New Monomer for the Synthesis of Intrinsic Antimicrobial Polymers with Enhanced Material Properties.

The use of biocidal compounds in polymers is steadily increasing because it is one solution to the need for safety and hygiene. It is possible to incorporate an antimicrobial moiety to a polymer. These polymers are referred to as intrinsic antimicrobial. The biocidal action results from contact of the polymer to the microorganisms, with no release of active molecules. This is particularly important in critical fields like food technology, medicine and ventilation technology, where migration or leaching is crucial and undesirable. The isomers N-(1,1-dimethylethyl)-4-ethenyl-benzenamine and N-(1,1-dimethyl-ethyl)-3-ethenyl-benzenamine (TBAMS) are novel (Co-)Monomers for intrinsic anti-microbial polymers. The secondary amines were prepared and polymerized to the corresponding water insoluble polymer. The antimicrobial activity was analyzed by the test method JIS Z 2801:2000. Investigations revealed a high antimicrobial activity against Staphylococcus aureus and Escherichia coli with a reduction level of >4.5 log10 units. Furthermore, scanning electron microscopy (SEM) of E. coli. in contact with the polymer indicates a bactericidal action which is caused by disruption of the bacteria cell membranes, leading to lysis of the cells.

Synthesis, characterization and antimicrobial activities of water-soluble amphiphilic copolymers containing ciprofloxacin and quaternary ammonium salts.

A series of novel water-soluble amphiphilic copolymers containing ciprofloxacin (CPF) and quaternary ammonium salts were prepared by the free-radical copolymerization of methacrylate monomers with CPF, quaternary ammonium salt (QAS) monomers and butyl acrylate (BA). The copolymers were characterized by 1H-NMR, charge density and UV measurements. The antimicrobial activities of the copolymers, based on the minimum inhibition concentration (MIC) against Escherichia coli (E. coli), varied with hydrophobic BA content in the copolymers. The incorporation of the antimicrobial moiety ciprofloxacin further enhanced the antibacterial activities of the copolymers, reducing the MIC to 4.0 ppm. The UV260 absorption measurements were applied to reveal the dynamic antimicrobial process of the copolymers. The results of inhibition zones and shaking flask methods of antimicrobial-modified fibers showed that the copolymers had excellent antimicrobial activities when applied to cellulose fibers. In the presence of 2 wt% (based on oven-dried fibers) of the copolymer, the growth inhibition reached 100%.

Self-decontaminating properties of fluorinated copolymers integrated with ciprofloxacin for synergistically inhibiting the growth of Escherichia coli

In this paper, copolymers composed of antibacterial monomer containing ciprofloxacin, methyl methacrylate (MMA), and 2-perfluorooctylethyl methacrylate (FMA) were prepared, and the surface properties and antibacterial performance of the copolymers and blends-mixed PMMA were investigated. Surface characterization using dynamic contact angle measurement and X-ray photoelectron spectroscopy showed that anti-adhesive fluorinated moieties and antimicrobial moieties were highly concomitant on the material surface. All the copolymers and blends films exhibited excellent antibacterial properties. It was found that the fluorinated antibacterial copolymers showed significantly enhanced antibacterial efficiency toward Escherichia coli bacterium, and even markedly prevented the formation of biofilm for long term. The PMMA films blended with fluorinated antibacterial polymer also show similar results. In contrast, the common copolymer without fluorinated units cannot effectively resist bacterial adhesion, proliferation, and prevent biofilm formation. The desirable antibacterial polymer prohibiting the biofilm formation performance of copolymer with special push-me/pull-you structure which weaken the interaction among polymer chains resulted in the more easy segregation of ciprofloxacin on surface in real environment by the help of synergistic effect of fluorinated units, potentially enabling the design of new self-decontaminating biomaterials for control biofouling.

Editorial of the Special Issue Antimicrobial Polymers

The special issue “Antimicrobial Polymers” includes research and review papers concerning the recent advances on preparation of antimicrobial polymers and their relevance to industrial settings and biomedical field. Antimicrobial polymers have recently emerged as promising candidates to fight microbial contamination onto surfaces thanks to their interesting properties. In this special issue, the main strategies pursued for developing antimicrobial polymers, including polymer impregnation with antimicrobial agents or synthesis of polymers bearing antimicrobial moieties, were discussed. The future application of these polymers either in industrial or healthcare settings could result in an extremely positive impact not only at the economic level but also for the improvement of quality of life.

English

pores thereby altering the membrane permeability, the nucleoside analogues which inhibit the synthesis of nucleic acids, some antibiotics (e.g. griseofulvin) which inhibit cell division by hampering the synthesis of the microtubules and others.1 Alternative and less toxic antifungal formulations may include natural products, synthetic agents and polymeric materials such as some saponins, alkaloids, peptides, essential Abstract Introduction Fungicidal assemblies can be built from lipids, polymers and/or drugs to yield optimal activity against fungus in virtual absence of haemolysis. Candida albicans has often been used as a model for testing novel fungicidal assemblies both in vitro and in vivo. Fungicidal drugs require appropriate formulations to improve their therapeutic index at low cost and toxicity. Inexpensive synthetic lipids or surfactants and biocompatible, water soluble polymers can be assembled to provide novel vehicles for carrying the fungicidal drugs or eventually being the fungicidal agent themselves. In this critical review, perspectives of some important fungicidal assemblies of low toxicity and cost are disclosed and major factors such as mobility of the quaternary ammonium moiety, charge of the fungus cell, size and shape of the fungicidal nanostructures determining their mode of action are presented. Conclusion The major factors determining the activity of fungicidal assemblies are the mobility of the antimicrobial moiety, the charge on the fungus cell and the size and shape of the antifungal assembly.