Activity and thermal stability improvements of glucose oxidase upon adsorption on core-shell PMMA-BSA nanoparticles.

The interaction and adsorption of enzyme, glucose oxidase (GOx), on poly(methyl methacrylate)-bovine serum albumin (PMMA-BSA) particles were studied by using a quartz crystal microbalance with dissipation (QCM-D) and laser light scattering (LLS). The enzyme was irreversibly immobilized on the PMMA-BSA particle surface. The amount of enzyme immobilized on PMMA-BSA particles and the enzymatic activity were determined by UV/vis measurements. The influences of pH and ionic strength on the adsorption indicate that the electrostatic interaction plays a major role on the immobilization. The adsorbed GOx can retain at least 80% of the free enzyme activity. Thermal stability studies reveal that the adsorbed GOx only losses 28% of its activity in comparison with a 64% activity loss of free GOx when it is incubated at 50 degrees C for 35 h.

Preparation of polypyrrole nanoparticles in reverse micelle and its application to glucose biosensor.

Polypyrrole nanoparticles were successfully synthesized in cetyltrimethyl ammonium bromide (CTAB)/hexanol/water reverse micelle. The morphology and particle size of the obtained nanoparticles were characterized with transmission electron microscope (TEM) and scanning electron microscopy (SEM). Glucose biosensors were formed with glucose oxidase (GOx) immobilized in conducting composite material consisting of polypyrrole nanoparticles and ethyl cellulose. The effects of reaction conditions such as molar ratio of polypyrrole nanoparticles to ethyl cellulose, working voltage, glucose concentration, temperature and solution pH on the electrochemical response of the GOx electrode were studied. Experimental results showed that the linear range of GOx electrode was 1.0 x 10(-6)-6 x 10(-3) mol/L and the detection limit was 1.0 x 10(-7) mol/L. The electrode exhibited fine repeatability and selectability, and its lifetime was greater than one month. AFM showed that the surface of conducting composite material-glucose oxidase electrode's presents uniform granular after washing paraffin wax with cyclohexane, which was favorable for enzyme-catalyzed reaction.

Construction of an implicit membrane environment for the lattice Monte Carlo simulation of transmembrane protein.

Due to the complexity of biological membrane, computer simulation of transmembrane protein's folding is challenging. In this paper, an implicit biological membrane environment has been constructed in lattice space, in which the lipid chains and water molecules were represented by the unoccupied lattice sites. The biological membrane was characterized with three features: stronger hydrogen bonding interaction, membrane lateral pressure, and lipophobicity index for the amino acid residues. In addition to the hydrocarbon core spanning region and the water solution, the lipid interface has also been represented in this implicit membrane environment, which was proved to be effective for the transmembrane protein's folding. The associated Monte Carlo simulations have been performed for SARS-CoV E protein and M2 protein segment (residues 18-60) of influenza A virus. It was found that the coil-helix transition of the transmembrane segment occurred earlier than the coil-globule transition of the two terminal domains. The folding process and final orientation of the amphipathic helical block in water solution are obviously influenced by its corresponding hydrophobicity/lipophobicity. Therefore, this implicit membrane environment, though in lattice space, can make an elaborate balance between different driving forces for the membrane protein's folding, thus offering a potential means for the simulation of transmembrane protein oligomers in feasible time.

Preparation of well-defined core-shell particles by Cu2+-mediated graft copolymerization of methyl methacrylate from bovine serum albumin.

Small well-defined core-shell poly(methyl methacrylate)-bovine serum albumin (PMMA-BSA) particles have been prepared in a direct one-step graft copolymerization of MMA from BSA at 75 degrees C in water with a trace amount of Cu2+ (5 microM). Initially, BSA generates free radicals and acts as a multifunctional macroinitiator, which leads to the formation of an amphiphilic PMMA-BSA grafting copolymer. Such formed copolymer chains act as a polymeric stabilizer to promote further emulsion polymerization of MMA inside, resulting in surfactant-free stable core-shell particles, confirmed by a transmission electron microscopic (TEM) analysis. The PMMA-BSA copolymers as well as PMMA homopolymer inside the particles were isolated by Soxhlet extraction and characterized by Fourier transform infrared spectroscopy (FT-IR) and thermogravimetry (TG). The highest grafting efficiency was approximately 80%. Effects of the reaction temperature, the MMA/BSA ratio, and the concentrations of Cu2+ and BSA on such core-shell particle formation have been systematically studied. Due to their inert PMMA core and biocompatible BSA shell, these small polymer particles are potentially useful in biomedical applications.