Piloting the membranolytic activities of peptides with a self-organizing map.

Antimicrobial peptides (AMPs) show remarkable selectivity toward lipid membranes and possess promising antibiotic potential. Their modes of action are diverse and not fully understood, and innovative peptide design strategies are needed to generate AMPs with improved properties. We present a de novo peptide design approach that resulted in new AMPs possessing low-nanomolar membranolytic activities. Thermal analysis revealed an entropy-driven mechanism of action. The study demonstrates sustained potential of advanced computational methods for designing peptides with the desired activity.

Efficient magnetic recycling of covalently attached enzymes on carbon-coated metallic nanomagnets.

In the pursuit of robust and reusable biocatalysts for industrial synthetic chemistry, nanobiotechnology is currently taking a significant part. Recently, enzymes have been immobilized on different nanoscaffold supports. Carbon coated metallic nanoparticles were found to be a practically useful support for enzyme immobilization due to their large surface area, high magnetic saturation, and manipulatable surface chemistry. In this study carbon coated cobalt nanoparticles were chemically functionalized (diazonium chemistry), activated for bioconjugation (N,N-disuccinimidyl carbonate), and subsequently used in enzyme immobilization. Three enzymes, ß-glucosidase, α-chymotrypsin, and lipase B were successfully covalently immobilized on the magnetic nonsupport. The enzyme-particle conjugates formed retained their activity and stability after immobilization and were efficiently recycled from milliliter to liter scales in short recycle times.

Organic synthesis on graphene.

Graphene is a two-dimensional crystalline carbon allotrope that has fascinated researchers worldwide and has extended the interest in carbon structures such as fullerenes and nanotubes. In this Account, we use electrical characterization tools to study chemistry on supported graphene. These experiments elucidate the way covalently bound phenyl units can change graphene's physical properties. Can we use chemistry to control electronic properties of graphene? What can we learn from well-known carbon allotropes like fullerenes? The surfaces of fullerenes and graphene show distinct differences in reactivity because of the high strain of sp² carbon in fullerenes compared with the complete lack of strain in graphene. Diazonium chemistry provides a versatile tool for attaching phenyl units covalently to carbon to produce advanced materials and electronic components, but diazonium-based carbon chemistry is strongly influenced by strain. Although fullerenes are highly reactive, graphite (stacks of graphene) remains relatively inert. We chemically introduce n- and p-like doping patterns in two-dimensional graphene using photolithography and extend the ability to chemically control doping to the chemical design of conducting and insulating areas. Thereby we can shape graphene surfaces into functional electronic devices. This Account also describes multistep synthesis on graphene-coated nanoparticles and the introduction of various functional groups on graphene surfaces. Only few functional groups can be produced directly via diazonium chemistry. To overcome this issue, we used these functional groups as starting points for more demanding organic reactions. We covalently attached chelating agents, catalysts, or polymers on the carbon surface. These more complex reactions facilitate the design of electronic modifications, intergraphene connections, and anchors for polymer incorporation. Diazonium chemistry forms strong covalent bridges between graphene and other areas of chemistry.

Incorporation of penicillin-producing fungi into living materials to provide chemically active and antibiotic-releasing surfaces.

Living materials: artificial biological niches are loaded with the penicillin-producing mold Penicillium chrysogenum. This living material consumes food through a nanoporous top layer and releases the antibiotic on-site. No reloading of the active compound is needed. Gram-positive bacteria were efficiently killed if nearby, whereas Gram-negative bacteria (control experiment, not sensitive to penicillin) were not affected.

Electrical resistivity of assembled transparent inorganic oxide nanoparticle thin layers: influence of silica, insulating impurities, and surfactant layer thickness.

The electrical properties of transparent, conductive layers prepared from nanoparticle dispersions of doped oxides are highly sensitive to impurities. Production of cost-effective thin conducting films for consumer electronics often employs wet processing such as spin and/or dip coating of surfactant-stabilized nanoparticle dispersions. This inherently results in entrainment of organic and inorganic impurities into the conducting layer leading to largely varying electrical conductivity. Therefore, this study provides a systematic investigation on the effect of insulating surfactants, small organic molecules and silica in terms of pressure dependent electrical resistivity as a result of different core/shell structures (layer thickness). Application of high temperature flame synthesis gives access to antimony-doped tin oxide (ATO) nanoparticles with high purity. This well-defined starting material was then subjected to representative film preparation processes using organic additives. In addition ATO nanoparticles were prepared with a homogeneous inorganic silica layer (silica layer thickness from 0.7 to 2 nm). Testing both organic and inorganic shell materials for the electronic transport through the nanoparticle composite allowed a systematic study on the influence of surface adsorbates (e.g., organic, insulating materials on the conducting nanoparticle's surface) in comparison to well-known insulators such as silica. Insulating impurities or shells revealed a dominant influence of a tunneling effect on the overall layer resistance. Mechanical relaxation phenomena were found for 2 nm insulating shells for both large polymer surfactants and (inorganic) SiO(2) shells.

Chemical modification of graphene characterized by Raman and transport experiments.

A chemical approach to modify the electronic transport of graphene is investigated by detailed transport and Raman spectroscopy measurements on Hall bar shaped samples. The functionalization of graphene with nitrobenzene diazonium ions results in a strong p-doping of the graphene samples and only slightly lower mobilities. Comparing Raman and transport data taken after each functionalization step allowed the conclusion that two preferential reactions take place on the graphene surface. In the beginning a few nitrobenzene molecules are directly attached to the graphene atoms creating defects. Afterwards these act as seeds for a polymer like growth not directly connected to the graphene atoms. The effects of solvents were excluded by thorough control measurements.

Incorporating microorganisms into polymer layers provides bioinspired functional living materials.

Artificial two-dimensional biological habitats were prepared from porous polymer layers and inoculated with the fungus Penicillium roqueforti to provide a living material. Such composites of classical industrial ingredients and living microorganisms can provide a novel form of functional or smart materials with capability for evolutionary adaptation. This allows realization of most complex responses to environmental stimuli. As a conceptual design, we prepared a material surface with self-cleaning capability when subjected to standardized food spill. Fungal growth and reproduction were observed in between two specifically adapted polymer layers. Gas exchange for breathing and transport of nutrient through a nano-porous top layer allowed selective intake of food whilst limiting the microorganism to dwell exclusively in between a confined, well-enclosed area of the material. We demonstrated a design of such living materials and showed both active (eating) and waiting (dormant, hibernation) states with additional recovery for reinitiation of a new active state by observing the metabolic activity over two full nutrition cycles of the living material (active, hibernation, reactivation). This novel class of living materials can be expected to provide nonclassical solutions in consumer goods such as packaging, indoor surfaces, and in biotechnology.

Two-layer membranes of calcium phosphate/collagen/PLGA nanofibres: in vitro biomineralisation and osteogenic differentiation of human mesenchymal stem cells.

The present study evaluates the in vitro biomedical performance of an electrospun, flexible, anisotropic bilayer with one layer containing a collagen to mineral ratio similar to that in bone. The double membrane consists of a poly(lactide-co-glycolide) (PLGA) layer and an amorphous calcium phosphate (a-CaP)/collagen (Col)/PLGA layer. In vitro biomineralisation and a cell culture study with human mesenchymal stem cells (hMSC) were conducted to characterise such membranes for possible application as biomaterials. Nanofibres with different a-CaP/Col/PLGA compositions were synthesised by electrospinning to mimic the actual composition of bone tissue. Immersion in simulated body fluid and in cell culture medium resulted in the deposition of a hydroxyapatite layer. Incubation of hMSC for 4 weeks allowed for assessment of the proliferation and osteogenic differentiation of the cells on both sides of the double membrane. Confocal laser scanning microscopy was used to observe the proper adhesion of the cells. Calcium and collagen content was proven by Alizarin red S and Sirius red assays. Acute cytotoxic effects of the nanoparticles or the chemicals used in the scaffold preparation could be excluded based on viability assays (alamarBlue and alkaline phosphatase activity). The findings suggest possible application of such double membranes is in treatment of bone defects with complex geometries as wound dressing material.

Selective chemical modification of graphene surfaces: distinction between single- and bilayer graphene.

Graphene modifications with oxygen or hydrogen are well known in contrast to carbon attachment to the graphene lattice. The chemical modification of graphene sheets with aromatic diazonium ions (carbon attachment) is analyzed by confocal Raman spectroscopy. The temporal and spatial evolution of surface-adsorbed species allows accurate tracking of the chemical reaction and identification of intermediates. The controlled transformation of sp(2) to sp(3) carbon proceeds in two separate steps. The presented derivatization is faster for single-layer graphene and allows controlled transformation of adsorbed diazonium reagents into covalently bound surface derivatives with enhanced reactivity at the edge of single-layer graphene. On bilayer graphene the derivatization proceeds to an adsorbed intermediate, which reacts slower to a covalently attached species on the carbon surface.

Magnetic EDTA: coupling heavy metal chelators to metal nanomagnets for rapid removal of cadmium, lead and copper from contaminated water.

Attachment of EDTA-like chelators to carbon coated metal nanomagnets results in a magnetic reagent for the rapid removal of heavy metals from solutions or contaminated water by three orders of magnitude to concentrations as low as microg L(-1).