Development of Silica Nanoparticle Supported Imprinted Polymers for Selective Lysozyme Recognition

Protein imprinted MIPs show notable potential for applications in many analytical areas such as clinical analysis, medical diagnostics and environmental monitoring, but also in drug delivery scenarios. In this study, we present various modifications of two different synthesis routes to create imprinted core-shell particles serving as a synthetic recognition material for the protein hen egg white (HEW) lysozyme. HEW lysozyme is used as food additive E 1105 for preservation due to its antibacterial effects. For facilitating quality and regulatory control analysis in food matrices, it is necessary to apply suitable isolation methods as potentially provided by molecularly imprinted materials. The highest binding capacity achieved herein was 58.82 mg/g with imprinting factors ranging up to 2.74, rendering these materials exceptionally suitable for selectively isolating HEW lysozyme.

The impact of folding modes and deuteration on the atomic resolution structure of hen egg-white lysozyme

The biological function of a protein is intimately related to its structure and dynamics, which in turn are determined by the way in which it has been folded. In vitro refolding is commonly used for the recovery of recombinant proteins that are expressed in the form of inclusion bodies and is of central interest in terms of the folding pathways that occur in vivo. Here, biophysical data are reported for in vitro-refolded hydrogenated hen egg-white lysozyme, in combination with atomic resolution X-ray diffraction analyses, which allowed detailed comparisons with native hydrogenated and refolded perdeuterated lysozyme. Distinct folding modes are observed for the hydrogenated and perdeuterated refolded variants, which are determined by conformational changes to the backbone structure of the Lys97–Gly104 flexible loop. Surprisingly, the structure of the refolded perdeuterated protein is closer to that of native lysozyme than that of the refolded hydrogenated protein. These structural differences suggest that the observed decreases in thermal stability and enzymatic activity in the refolded perdeuterated and hydrogenated proteins are consequences of the macromolecular deuteration effect and of distinct folding dynamics, respectively. These results are discussed in the context of both in vitro and in vivo folding, as well as of lysozyme amyloidogenesis.