Synthetic polymer-based antimicrobial materials destroy conventional antibiotic resistant microorganisms. Although these antibacterial polymers imitate the properties of antimicrobial peptides (AMPs), their effect on bacterial cell morphology has not been studied in detail. To investigate the morphology change of a bacterial cell in the presence of antimicrobial polymer, herein we have designed and synthesized side-chain amino acid-based cationic polymers, which showed efficient antibacterial activity against Gram-negative (Escherichia coli), as well as Gram-positive (Bacillus subtilis) bacteria. Morphological switching from a rod shape to a spherical shape of E. coli cells was observed by field emission-scanning electron microscopy analysis due to cell wall disruption, whereas the B. subtilis cell structure and size remained intact, but stacks of the cells formed after polymer treatment. The zone of inhibition experiment on an agar plate for E. coli cells exhibited drastic morphological changes at the vicinity of the polymer-treated portion and somewhat less of an effect at the periphery of the plate.
β-Glucosidase (BG) catalyzes the hydrolysis of cellobiose to glucose and is a rate-limiting enzyme in the conversion of lignocellulosic biomass to sugars toward biofuels. Since the cost of enzyme is a major contributor to biofuel economics, we report the bioconjugation of a temperature-responsive polymer with the highly active thermophilic β-glucosidase (B8CYA8) from Halothermothrix orenii toward improving enzyme recyclability. The bioconjugate, with a lower critical solution temperature (LCST) of 33 °C withstands high temperatures up to 70 °C. Though the secondary structure of the enzyme in the conjugate is slightly distorted with a higher percentage of β-sheet like structure, the stability and specific activity of B8CYA8 in the conjugate remains unaltered up to 30 °C and retains more than 70% specific activity of the unmodified enzyme at 70 °C. The conjugate can be reused for β-glucosidic bond cleavage of cellobiose for at least four cycles without any significant loss in specific activity.
A single step process of integrating a resonantly tuned silver nanoparticle into photonic crystal nanobeam cavities fabricated by focused ion beam milling is presented. Even though the quality factor of the cavities is reduced by a factor of 20, the emission peak at the cavity resonance is enhanced by 5-fold with respect to the cavities without the metal nanoparticle. The fluorescence is also compared before and after etching away the nanoparticle. Experimental quality factors and wavelength shifts are found to agree reasonably well with simulation values. These results are promising for future single photon emission studies involving the incorporation of quantum dot or NV center emitters into hybrid plasmonic/photonic crystal cavities for enhanced emission rates while retaining reasonably high quality factors.
To evaluate the comparative antibacterial activity of leucine-based cationic polymers having linear, hyperbranched, and star architectures containing both hydrophilic and hydrophobic segments against Gram-negative bacterium, Escherichia coli (E. coli), herein we performed zone of inhibition study, minimum inhibitory concentration (MIC) calculation, and bacterial growth experiment. The highest antibacterial activity in terms of the MIC value was found in hyperbranched and star architectures because of the greater extent of cationic and hydrophobic functionality, enhancing cell wall penetration ability compared to that of the linear polymer. The absence of the bacterial regrowth stage in the growth curve exhibited the highest bactericidal capacity of star polymers, when untreated cells (control) already reached to the stationary phase, whereas the bacterial regrowth stage with a delayed lag phase was critically observed for linear and hyperbranched architectures displaying lower bactericidal efficacy. Coagulation of E. coli cells, switching of cell morphology from rod to sphere, and lengthening due to stacking in an antimicrobial polymer-treated environment at the bacterial regrowth stage in liquid media were visualized critically by field emission scanning electron microscopy and confocal fluorescence microscopy instruments in the presence of 4′,6-diamidino-2-phenylindole stain.
The design of a new drug material through modification of some well-known antibiotics to combat pathogenic bacteria must include a complete understanding of matrix regulation because the human body consists of primarily three types of matrices, that is, solid, semisolid, and liquid, all of which have a tendency to regulate antibacterial efficacy along with the bactericidal mechanism of several antimicrobial agents. Herein, matrix-dependent action of ciprofloxacin-based polymeric hydrogel scaffold was explored against a new species of Vibrio, namely, Vibrio chemaguriensis Iso1 (V. chemaguriensis), which is resistant to most of the common antibiotics and possess genes that can be linked to pathogenicity. Ciprofloxacin was attached to the polymeric system consisting of hydrophilic polyethylene glycol methyl ether methacrylate (PEGMA) and zwitterionic sulphabetaine methacrylate (SBMA) with an antifouling nature via covalent linkage leading to effective polymer antibiotic conjugates (PACs) with linear and hyperbranched architectures. The hyperbranched PAC was transformed to a polymeric gel exhibiting greater antibacterial efficacy in solid matrix than that of the liquid one with a complete bactericidal effect and rod to spherical switching of bacterial cell followed by chain formation via "dual" contact activity and release mechanism through sustained removal of thiol-terminated ciprofloxacin fragment along with an equilibrium swelling and deswelling process. Lower killing efficacy was displayed in the liquid matrix with an intact cell morphology that is due to lack of forced contact between the cell wall and gel surface as well as entrapment of released bioactive fragment via an additional thick exopolysaccharide (EPS) layer, which represents a great challenge to modern medical sciences.
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