Boronic Acid Modified Polymer Nanoparticles for Enhanced Bacterial Deactivation.

A new method has been developed to enhance the antibacterial efficiency of traditional antibiotics. Chloramphenicol-imprinted polymer particles were decorated with boronic acid to improve their binding to both Gram-negative and -positive bacteria. The polymer particles have a high antibiotic loading and provide a slow release of the antibiotic payload to deactivate the target bacteria. The boronic acid modified polymer particles not only contribute to enhanced antibacterial efficiency, but also have the potential to act as scavengers to remove unused antibiotic from the environment.

Recyclable nanoparticles based on a boronic acid-diol complex for the real-time monitoring of imprinting, molecular recognition and copper ion detection.

Molecularly imprinted polymers (MIPs) have now become one of the most remarkable materials in the field of molecular recognition. Although many efforts have been made to study the process and mechanism of molecular imprinting, it has not been possible to monitor the interactions between the template and the growing polymer chains under real-time experimental conditions. The behavior of the template-monomer complex during the whole polymerization process has remained largely unknown. In this work, we introduce a fluorescence technique that allows monitoring of the template-functional monomer complex during an actual imprinting process, as well as the real-time signaling of template binding and dissociation from the imprinted polymer. For the first proof-of-principle, we select Alizarin Red S (ARS) and 4-vinylphenylboronic acid as the template and functional monomer, respectively, to synthesize MIP particles via precipitation polymerization. As the formation of the template-functional monomer complex leads to strong fluorescence emission, it allows the status of the template binding to be monitored throughout the whole reaction process in real time. Using the same fluorescence technique, the kinetics of template binding and dissociation can be studied directly without particle separation. The hydrophilic MIP particles can be used as a scavenger to remove ARS from water. In addition, the MIP particles can be used as a recyclable sensor to detect Cu ions. As the Cu ion forms a stable complex with ARS, it causes ARS to dissociate from the MIP nanoparticles, leading to effective fluorescence quenching. The non-separation analytical method based on fluorescence measurement provides a convenient means to study molecular imprinting reactions and the kinetics of molecular recognition using imprinted polymers. The recyclable nanoparticle sensor allows toxic Cu ions to be detected directly in water in the range of 0.1-100 µM with a recovery of 84-95%.

Biotechnological Approaches on Engineering Medicinal Tropane Alkaloid Production in Plants.

Hyoscyamine and scopolamine, belonging to medicinal tropane alkaloids (MTAs), are potent anticholinergic drugs. Their industrial production relies on medicinal plants, but the levels of the two alkaloids are very low in planta. Engineering the MTA's production is an everlasting hot topic for pharmaceutical industry. With understanding the MTA's biosynthesis, biotechnological approaches are established to produce hyoscyamine and scopolamine in an efficient manner. Great advances have been obtained in engineering MTA's production in planta. In this review, we summarize the advances on the biosynthesis of MTAs and engineering the MTA's production in hairy root cultures, as well in plants. The problems and perspectives on engineering the MTA's production are also discussed.

Imprinted Polymer Beads Loaded with Silver Nanoparticles for Antibacterial Applications.

After the emergence of multidrug-resistant strains, antibiotic resistance in bacteria has become an important problem. Thus, materials for combating multidrug-resistant bacteria are of vital importance. In this work, we developed an antibacterial material that can selectively capture and destruct bacteria on the basis of their physical characteristics. To achieve bacterial capture and deactivation with a single material, we used bacterial cells as templates to synthesize surface-imprinted polymer beads in bacteria-stabilized Pickering emulsions. Acrylate-functionalized polyethylenimine was used to coat the bacterial surface so that the coated bacteria can act as a particle stabilizer to establish an oil-in-water Pickering emulsion. Hydrophobic Ag nanoparticles were introduced into the oil phase composed of cross-linking monomers. Bacteria-imprinted beads (BIB) were obtained after the oil phase was polymerized. Bacterial binding experiments confirmed the importance of the imprinted sites for specific recognition with the target bacteria. The Ag nanoparticles embedded inside the polymer beads enhanced bacterial inactivation and reduced the leakage of heavy metal in aquatic environment. The combination of bacteria-imprinting with delivery of general-purpose antibacterial reagents offers a useful approach toward selective capture and destruction of bacteria.

Synthesis of Bacteria Imprinted Polymers by Pickering Emulsion Polymerization.

Molecularly imprinted polymers have been studied for a long time and have found useful applications in many fields. In most cases, small organic molecules are used as templates to synthesize imprinted polymers. In contrast to low molecular weight targets, large biological molecules and cells are more challenging to use as templates to synthesize cell-recognizing materials. This chapter presents an interfacial imprinting method to synthesize bacteria-recognizing polymer beads using Pickering emulsion polymerization. The tendency of bacteria to reside between two immiscible liquids is utilized to create surface-imprinted binding sites on cross-linked polymer microspheres.

Boronic Acid Functionalized Nanosilica for Binding Guest Molecules.

Dendritic fibrous nanosilica (DFNS) has very high surface area and well-defined nanochannels; therefore, it is very useful as supporting material for numerous applications including catalysis, sensing, and bioseparation. Due to the highly restricted space, addition of molecular ligands to DFNS is very challenging. This work studies how ligand conjugation in nanoscale pores in DFNS can be achieved through copper-catalyzed click reaction, using an optional, in situ synthesized, temperature-responsive polymer intermediate. A clickable boronic acid is used as a model to investigate the ligand immobilization and the molecular binding characteristics of the functionalized DFNS. The morphology, composition, nanoscale pores, and specific surface area of the boronic acid functionalized nanosilica were characterized by electron microscopy, thermogravimetric and elemental analysis, Fourier transform infrared spectroscopy, and nitrogen adsorption-desorption measurements. The numbers of boronic acid molecules on the modified DFNS with and without the polymer were determined to be 0.08 and 0.68 mmol of ligand/g of DFNS, respectively. We also studied the binding of small cis-diol molecules in the nanoscale pores of DFNS. The boronic acid modified DFNS with the polymer intermediate exhibits higher binding capacity for Alizarin Red S and nicotinamide adenine dinucleotide than the polymer-free DFNS. The two types of boronic acid modified DFNS can bind small cis-diol molecules in the presence of large glycoproteins, due in large part to the effect of size exclusion provided by the nanochannels in the DFNS.

Preparation of Boronic Acid-Functionalized Cryogels Using Modular and Clickable Building Blocks for Bacterial Separation.

Composite cryogels containing boronic acid ligands are synthesized for effective separation and isolation of bacteria. The large and interconnected pores in cryogels enable fast binding and release of microbial cells. To control bacterial binding, an alkyne-tagged boronic acid ligand is conjugated to azide-functionalized cryogel via the Cu(I)-catalyzed azide-alkyne cycloaddition reaction. The boronic acid-functionalized cryogel binds Gram-positive and Gram-negative bacteria through reversible boronate ester bonds, which can be controlled by pH and simple monosaccharides. To increase the capacity of affinity separation, a new approach is used to couple the alkyne-tagged phenylboronic acid to cryogel via an intermediate polymer layer that provides multiple immobilization sites. The morphology and chemical composition of the composite cryogel are characterized systematically. The capability of the composite cryogel for the separation of Gram-positive and Gram-negative bacteria is investigated. The binding capacities of the composite cryogel for Escherichia coli and Staphylococcus epidermidis are 2.15 × 109 and 3.36 × 109 cfu/g, respectively. The bacterial binding of the composite cryogel can be controlled by adjusting pH. The results suggest that the composite cryogel may be used as affinity medium for rapid separation and isolation of bacteria from complex samples.

Photoconjugation of temperature- and pH-responsive polymer with silica nanoparticles for separation and enrichment of bacteria.

A new photoconjugation approach was developed to prepare nanoparticle-supported boronic acid polymer for effective separation and enrichment of bacteria. The photo-activated polymer immobilization was demonstrated by coupling an azide-modified copolymer of N-isopropylacrylamide and glycidyl methacrylate to a perfluorophenyl azide-modified silica surface. The thermoresponsive polymer was synthesized using reversible addition fragmentation chain transfer polymerization followed by conversion of the pendant epoxides into azide groups. The perfluorophenyl azide-modified silica nanoparticles were synthesized by an amidation reaction between amino-functionalized silica and pentafluorobenzoyl chloride, and a subsequent treatment with sodium azide. Bacteria-capturing boronic acid was conjugated to the silica-supported polymer chains via Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC) click reaction. The particle size, morphology and organic content of the composite nanoparticles were characterized systematically. The capability of the nanocomposite to bind Gram-positive and Gram-negative bacteria was investigated. The nanocomposite exhibited high binding capacities for E. coli (13.4 × 107 CFU/mg) and S. epidermidis (7.66 × 107 CFU/mg) in phosphate buffered saline. The new photoconjugation strategy enables fast and straightforward grafting of functional polymers on surface, which opens many new opportunities for designing functional materials for bioseparation and biosensing.

Iron-chelated thermoresponsive polymer brushes on bismuth titanate nanosheets for metal affinity separation of phosphoproteins.

Separation of phosphoproteins plays an important role for identification of biomarkers in life science. In this work, bismuth titanate supported, iron-chelated thermoresponsive polymer brushes were prepared for selective separation of phosphoproteins. The iron-chelated thermoresponsive polymer brushes were synthesized by surface-initiated atom transfer radical polymerization of N-isopropylacrylamide and glycidyl methacrylate, followed by a ring opening reaction of epoxy group, and chelation of the obtained cis-diols with Fe3+ ions. The composite material was characterized to determine the size and thickness, the content of the organic polymer and the metal loading. The bismuth titanate supported, iron-chelated thermoresponsive polymer brushes showed selective binding for phosphoproteins in the presence of abundant interfering proteins, and a high binding capacity for phosphoproteins by virtue of the metal affinity between the metal ions on the polymer brushes and the phosphate groups in the phosphoproteins (664 mg ß-Casein per g sorbent). The thermoresponsive property of the polymer brushes made it possible to adjust phosphoprotein binding by changing temperature. Finally, separation of phosphoproteins from a complex biological sample (i.e. milk) was demonstrated using the nanosheet-supported thermoresponsive polymer brushes.

Nanoparticle-supported polymer brushes for temperature-regulated glycoprotein separation: investigation of structure-function relationship.

In this work, we synthesized a series of nanoparticle-supported boronic acid polymer brushes for affinity separation of glycoproteins. Polymer brushes were prepared by surface-initiated atom transfer radical polymerization of glycidyl methacrylate and N-isopropylacrylamide, followed by stepwise modification of the pendant as well as the end functional groups to introduce boronic acid moieties through a Cu(i)-catalyzed alkyne-azide cycloaddition reaction. We investigated the impact of the polymer structure on glycoprotein binding under different pH and temperature conditions, and established new methods that allow glycoproteins to be more easily isolated and recovered with minimal alteration in solvent composition. Our experimental results suggest that for the separation of glycoproteins, terminal boronic acids located at the end of polymer chains play the most important role. The thermo-responsibility of the new affinity adsorbents, in addition to the high capacity for glycoprotein binding (120 mg ovalbumin per g adsorbent), provides a convenient means to realize simplified bioseparation not only for glycoproteins, but also for other carbohydrate-containing biological molecules.

Dynamic assembly of molecularly imprinted polymer nanoparticles.

Manipulation of specific binding and recycling of materials are two important aspects for practical applications of molecularly imprinted polymers. In this work, we developed a new approach to control the dynamic assembly of molecularly imprinted nanoparticles by surface functionalization. Molecularly imprinted polymer nanoparticles with a well-controlled core-shell structure were synthesized using precipitation polymerization. The specific binding sites were created in the core during the first step imprinting reaction. In the second polymerization step, epoxide groups were introduced into the particle shell to act asan intermediate linker to immobilize phenylboronic acids, as well as to introduce cis-diol structures on surface. The imprinted polymer nanoparticles modified with boronic acid and cis-diol structures maintained high molecular binding specificity, and the nanoparticles could be induced to form dynamic particle aggregation that responded to pH variation and chemical stimuli. The possibility of modulating molecular binding and nanoparticle assembly in a mutually independent fashion can be exploited in a number of applications where repeated use of precious nanoparticles is needed.

Individually immobilized and surface-modified hydrocarbon-degrading bacteria for oil emulsification and biodegradation.

Effective emulsification plays an important role in the treatment of marine oil spills. The negative effects of chemical surfactants have necessitated a search for alternative dispersant that are sustainable and environmentally-friendly. To identify alternate dispersants, oil-in-seawater emulsions stabilized by hydrocarbon-degrading bacteria were investigated. After individual immobilization and surface-modification, the hydrocarbon-degrading bacteria, Bacillus cereus S-1, was found to produce a stable oil-in-seawater Pickering emulsion, which was similar to particle emulsifiers. The individual immobilization and surface-modification process improved the surface hydrophobicity and wettability of the bacterial cells, which was responsible for their effective adsorption at the oil-water interface. Through effective emulsification, the biodegradation of oil was remarkably facilitated by these treated bacteria, because of the increased interfacial area. By combining the emulsification and biodegradation, the results of this reported work demonstrated a novel approach for developing environmentally-friendly bioremediation technology in the field of oil treatment.