Deniz Aktaş Uygun

Lysozyme-Based Antibacterial Nanomotors

An effective and rapid bacterial killing nanotechnology strategy based on lysozyme-modified fuel-free nanomotors is demonstrated. The efficient antibacterial property of lysozyme, associated with the cleavage of glycosidic bonds of peptidoglycans present in the bacteria cell wall, has been combined with ultrasound (US)-propelled porous gold nanowire (p-AuNW) motors as biocompatible dynamic bacteria nanofighters. Coupling the antibacterial activity of the enzyme with the rapid movement of these p-AuNWs, along with the corresponding fluid dynamics, promotes enzyme-bacteria interactions and prevents surface aggregation of dead bacteria, resulting in a greatly enhanced bacteria-killing capability. The large active surface area of these nanoporous motors offers a significantly higher enzyme loading capacity compared to nonporous AuNWs, which results in a higher antimicrobial activity against Gram-positive and Gram-negative bacteria. Detailed characterization studies and control experiments provide useful insights into the underlying factors controlling the antibacterial performance of the new dynamic bacteria nanofighters. Rapid and effective killing of the Gram-positive Micrococcus lysodeikticus bacteria (69-84% within 1-5 min) is demonstrated.

Micromotor-Based Biomimetic Carbon Dioxide Sequestration: Towards Mobile Microscrubbers.

We describe a mobile CO2 scrubbing platform that offers a greatly accelerated biomimetic sequestration based on a self-propelled carbonic anhydrase (CA) functionalized micromotor. The CO2 hydration capability of CA is coupled with the rapid movement of catalytic micromotors, and along with the corresponding fluid dynamics, results in a highly efficient mobile CO2 scrubbing microsystem. The continuous movement of CA and enhanced mass transport of the CO2 substrate lead to significant improvements in the sequestration efficiency and speed over stationary immobilized or free CA platforms. This system is a promising approach to rapid and enhanced CO2 sequestration platforms for addressing growing concerns over the buildup of greenhouse gas.

Self-propelled chelation platforms for efficient removal of toxic metals

Water-powered ligand-modified micromotors that offer efficient ‘on-the-fly’ chelation of heavy metal contaminants are described. The Janus micromotors are prepared by functionalizing Mg/Au microsphere motors with a self-assembled monolayer of meso-2,3-dimercaptosuccinic acid (DMSA). The resulting micromotors propel autonomously in complex environmental and biological matrices, containing chloride and surfactant, obviating the need for external (peroxide) fuel or expensive Pt catalysts. Such self-propelled micromotors act as highly efficient dynamic chelation platforms that offer significantly shorter and more efficient water remediation processes compared to the common use of static remediation agents. The effective decontamination capability of the DMSA-based Janus micromotors is illustrated towards the rapid removal of Zn(II), Cd(II) and Pb(II). Factors influencing the micromachine-enhanced metal chelation process, such as the navigation time and number of motors, have been investigated. High removal efficiencies of ∼100% are obtained for all target metals following 2 min treatment of serum, seawater or lakewater samples spiked with 500 μg L−1 of each heavy metal. The chelation mechanism has been characterized using the Langmuir model, indicating strong interaction and monolayer-type adsorption of the target heavy metals onto the DMSA-binding layer. The new nanomotor concept holds considerable promise towards future metal remediation applications.

Dye Attached Nanoparticles for Lysozyme Adsorption

In this work, Reactive Blue 15 dye functionalized poly(HEMA) nanoparticles were synthesized for reversible adsorption of lysozyme from its aqueous solution. For this, nano-sized poly(HEMA) nanoparticles were synthesized by the surfactant free emulsion polymerization. Reactive Blue 15 dye then covalently attached to the polymeric structure. These novel dye attached poly(HEMA) nanoparticles were used for the adsorption of lysozyme. Characterization of dye attached nanoparticles was carried out by using FTIR, AFM, and elemental analysis. Incorporation of the dye onto the polymeric structure was demonstrated by FTIR and elemental analysis, while the size and the shape of the nanoparticles were shown by AFM. The incorporated amount of the dye was found to be 70.3 μmol/g nanoparticle with sulphur stoichiometry and it was found that the prepared nanoparticles were in a spherical form and were about 100 nm diameter. Lysozyme adsorption studies were carried out with different conditions (pH, lysozyme concentration, temperature, and ionic strength) and maximum lysozyme adsorption was found to be 630.7 mg/g nanoparticle at pH 7.0 in 25°C medium temperature. Adsorbed lysozyme was desorbed by 1.0 M of NaCl with 96% recovery and synthesized dye-attached nanoparticles were used 10 times without any decrease in their adsorption capacity.

Synthesis and characterization of amino acid containing Cu(II) chelated nanoparticles for lysozyme adsorption.

Immobilized metal ion affinity chromatography (IMAC) is a useful method for adsorption of proteins that have an affinity for transition metal ions. In this study, poly(hydroxyethyl methacrylate-methacryloyl-L-tryptophan) (PHEMATrp) nanoparticles were prepared by surfactant free emulsion polymerization. Then, Cu(II) ions were chelated on the PHEMATrp nanoparticles to be used in lysozyme adsorption studies in batch system. The maximum lysozyme adsorption capacity of the PHEMATrp nanoparticles was found to be 326.9 mg/g polymer at pH 7.0. The nonspecific lysozyme adsorption onto the PHEMA nanoparticles was negligible. In terms of protein desorption, it was observed that adsorbed lysozyme was readily desorbed in medium containing 1.0 M NaCl. The results showed that the metal-chelated PHEMATrp nanoparticles can be considered as a good adsorbent for lysozyme purification.

Controlled release of curcumin from poly(HEMA-MAPA) membrane

Abstract In this work, poly(HEMA-MAPA) membranes were prepared by UV-polymerization technique. These membranes were characterized by SEM, FTIR, and swelling studies. Synthesized membranes had high porous structure. These membranes were used for controlled release of curcumin which is already used as folk remedy and used as drug for some certain diseases and cancers. Curcumin release was investigated for various pHs and temperatures. Optimum drug release yield was found to be as 70% at pH 7.4 and 37 °C within 2 h period. Time-depended release of curcumin was also investigated and its slow release from the membrane demonstrated within 48 h.

Dye functionalized cryogel columns for reversible lysozyme adsorption

In this study, poly (methyl methacrylate–glycidyl methacrylate) [poly(MMA-GMA)] cryogels were prepared by radical cryocopolymerization of MMA with GMA as a functional comonomer. Reactive Green 19 dye was then attached to the cryogel by nucleophilic substitution reaction, and this dye-attached cryogel column was used for lysozyme adsorption. Characterization of the cryogel was performed by Fourier transform infrared spectroscopy, environmental scanning electron microscopy, Brunauer–Emmett–Teller, and energy dispersive X-ray analysis. Pore size of the cryogels was 15–30 μm and pores were interconnected structure. Attached amount of Reactive Green 19 to cryogel support was calculated as 106.25 μmol/g cryogel. Lysozyme adsorption studies were carried out by using a continuous system. It was found that the maximum amount of lysozyme adsorption (32 mg/g cryogel) obtained from experimental results was found to be approximately same with the calculated Langmuir adsorption capacity (33 mg/g cryogel). Desorption of adsorbed lysozyme was carried out by using 1.5 M NaCl in pH 4.5 acetate buffer, and desorption yield was found to be 97.4%. Cryogels were very stable, and it was found that there was no remarkable reduction in the adsorption capacity at the end of ten adsorption–desorption cycles. As a result, Reactive Green 19-attached cryogels have great advantages such as easy preparation, rapid adsorption, and desorption, being economic and allowing the direct separation of proteins.

A novel support for antibody purification: fatty acid attached chitosan beads.

Linoleic acid attached chitosan beads [poly(LA-Ch)] (1.25 microm in diameter) are obtained by the formation of amide linkages between linoleic acid and chitosan. Poly(LA-Ch) beads are characterized by FTIR, TEM, and swelling studies. Poly(LA-Ch) beads are used for the purification of immunoglobulin-G (IgG) from human plasma in a batch system. The maximum IgG adsorption is observed at pH 7.0 for HEPES buffer. IgG adsorption onto the plain chitosan beads is found to be negligible. Adsorption values up to 136.7 mg/g from aqueous solutions are obtained by poly(LA-Ch) beads. IgG adsorption saw an increase as a result of increasing temperature. Higher amounts of IgG are adsorbed from human plasma (up to 390 mg/g) with a purity of 92%. The adsorption phenomena appeared to follow a typical Langmuir isotherm. It is observed that IgG could be repeatedly adsorbed and desorbed without significant loss when we take into account the adsorption amount. It is concluded that the poly(LA-Ch) beads allowed one-step purification of IgG from human plasma.

Immobilization of amyloglucosidase onto macroporous cryogels for continuous glucose production from starch

Poly(methyl methacrylate-glycidyl methacrylate) [Poly(MMA-GMA)] cryogels were synthesized using monomers of methylmethacrylic acid and epoxy group bearing GMA via radical cryopolymerization technique. Synthesized cryogels were used for the immobilization of amyloglucosidase to the cryogel surface using epoxy chemistry. Characterizations of the free and immobilized amyloglucosidase were carried out by comparing the optimum and kinetic parameters of enzymes. For this, pH and temperature profiles of free and immobilized preparation were studied and, it was found that, optimum pH of enzyme was not change upon immobilization (pH 5.0), while optimum temperature of the enzyme shifted 10 °C to warmer region after immobilization (optimum temperatures for free and immobilized enzyme were 55 and 65 °C, respectively). Kinetic parameters of free and immobilized enzyme were also investigated and Km values of free and immobilized amyloglucosidase were found to be 2.743 and 0.865 mg/mL, respectively. Vmax of immobilized amyloglucosidase was found to be (0.496 µmol/min) about four times less than that of free enzyme (2.020 µmol/min). Storage and operational stabilities of immobilized amyloglucosidase were also studied and it was showed that immobilized preparation had much more stability than free preparation. In the present work, amyloglucosidase immobilized poly(MMA-GMA) cryogels were used for continuous glucose syrup production from starch for the first time. Efficiency of immobilized enzyme was investigated and released amount of glucose was found to be 2.54 mg/mL at the end of the 5 min of hydrolysis. The results indicate that the epoxy functionalized cryogels offer a good alternative for amyloglucosidase immobilization applications with increased operational and thermal stability, and reusability. Also, these cryogels can be used for immobilization of other industrially valuable enzymes beyond amyloglucosidase.

Immobilization of alcohol dehydrogenase onto metal-chelated cryogels

In this presented work, poly(HEMA–GMA) cryogel was synthesized and used for the immobilization of alcohol dehydrogenase. For this, synthesized cryogels were functionalized with iminodiacetic acid and chelated with Zn2+. This metal-chelated cryogels were used for the alcohol dehydrogenase immobilization and their kinetic parameters were compared with free enzyme. Optimum pH was found to be 7.0 for both immobilized and free enzyme preparations, while temperature optima for free and immobilized alcohol dehydrogenase was 25 °C. Kinetic constants such as Km, Vmax, and kcat for free and immobilized form of alcohol dehydrogenase were also investigated. kcat value of free enzyme was found to be 3743.9 min−1, while kcat for immobilized enzyme was 3165.7 min−1. Thermal stability of the free and immobilized alcohol dehydrogenase was studied and stability of the immobilized enzyme was found to be higher than free form. Also, operational stability and reusability profile of the immobilized alcohol dehydrogenase were investigated. Finally, storage stability of the free and immobilized alcohol dehydrogenase was studied, and at the end of the 60 days storage, it was demonstrated that, immobilized alcohol dehydrogenase was exhibited high stability than that of free enzyme.