Weak protein-nanoparticle (NP) interactions are studied in a low binding regime as a model for the soft protein corona around nanoparticles in complex biological fluids. Noncovalent, reversible interactions between Subtilisin Carlsberg (SC) and silica NPs shows significant alteration in conformation and enzymatic activity in a NP-size dependent manner. Very weak interactions between SC and silica NPs were revealed by centrifugation-based separations and further supported by small-angle X-ray scattering, while bovine serum albumin was used as a strongly interacting reference. Secondary and tertiary structure changes of SC were studied via circular dichroism and correlated to enzymatic activity where the enzyme kinetics showed a critical role for nanoparticle size.

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Three dimensional hierarchical materials are widespread in nature but are difficult to synthesize by using self-assembly/organization. Here, we employ a gel-liquid interface to obtain centimeter-long ∼100 μm diameter tubes with complex mineral wall structures that grow from the interface into solution. The gel, made from gelatin, is loaded with metal chloride salt, whereas the solution is a high pH anion source. Tubes were obtained with a range of cations (Ca(2+) , Sr(2+) , Ba(2+) , Cu(2+) , and Zn(2+) ) and anions (CO3 (2-) and PO4 (3-) ). The crystalline phases found in the tube walls corresponded to expectations from solution chemistries and phase solubilities. The growth mechanism is found to be akin to that of chemical gardens. The divalent cations modify the strength of the gelatin gel in a manner that involves not only simple electrostatic screening, but also ion-specific effects. Thus, tubes were not obtained for those ions and/or concentrations that significantly changed the gel's mechanical structure. At high Cu(2+) loading, for example, vertical convection bands, not Liesegang bands, were observed in the gels.

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Nanostructured materials strongly modulate the behavior of adsorbed proteins; however, the characterization of such interactions is challenging. Here we present a novel method combining protein adsorption studies at nanostructured quartz crystal microbalance sensor surfaces (QCM-D) with optical (surface plasmon resonance SPR) and electrochemical methods (cyclic voltammetry CV) allowing quantification of both bound protein amount and activity. The redox enzyme glucose oxidase is studied as a model system to explore alterations in protein functional behavior caused by adsorption onto flat and nanostructured surfaces. This enzyme and such materials interactions are relevant for biosensor applications. Novel nanostructured gold electrode surfaces with controlled curvature were fabricated using colloidal lithography and glancing angle deposition (GLAD). The adsorption of enzyme to nanostructured interfaces was found to be significantly larger compared to flat interfaces even after normalization for the increased surface area, and no substantial desorption was observed within 24 h. A decreased enzymatic activity was observed over the same period of time, which indicates a slow conformational change of the adsorbed enzyme induced by the materials interface. Additionally, we make use of inherent localized surface plasmon resonances in these nanostructured materials to directly quantify the protein binding. We hereby demonstrate a QCM-D-based methodology to quantify protein binding at complex nanostructured materials. Our approach allows label free quantification of protein binding at nanostructured interfaces.

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Protein‐nanoparticle (NP) interactions offer a powerful tool to modulate enzymatic activity and activate biocatalysis. However, strong scattering effects may complicate conventional spectrophotometric studies of NP‐assisted biocatalysis. Here, bioelectrocatalytic oxidation of cellobiose by the Humicola insolens cellobiose dehydrogenase (CDH) was electrochemically interrogated with two redox mediators, ferricyanide and cytochrome c, and two types of carbon NPs (CNPs), unmodified (CNP) and negatively charged 4‐sulfophenyl‐modified (CNP‐) ones. We show that both CNPs activate the bioelectrocatalytic oxidation of cellobiose and improve the enzymatic specificity reflected in the kf/KM relationship of 6680 (no CNPs), 15500 (CNP‐) and 10444 M−1 s−1 (CNP) shown for ferricyanide‐mediated and 1262 (no CNPs), 3630 (CNP‐), and 6375 M−1 s−1 (CNP) for cytochrome c‐mediated reactions. The bioelectrocatalysis activation by CNPs apparently results from the interfacial/conformational regulation of the CDH activity and the CDH‐electron acceptor reactivity, which may also be the case of other nanomaterials such as frequently used in bioelectrochemistry carbon nanotubes.