TERMINUS WP4 Enzyme immobilization, protection, and triggering. TASK 4.1 Experimental data – Biocatalytic red-ox systems based on laccases

Abstract Laccases (EC 1.10.3.2) are multi-copper oxidoreductase metalloenzymes found in various organisms, including bacteria, fungi, and plants. Laccases typically catalyze the oxidation of phenolic compounds along with the reduction of molecular oxygen to water. In TERMINUS WP4, task 4.1, we strive to improve the thermostability of the laccase enzyme from Pleurotus ostreatus (received from Biopox Srl). Our enzyme protection strategy is based on a method developed by FHNW that consists of shielding enzymes immobilized at the surface of silica-based nanoparticles within a soft organosilica layer. The components of this protective shield, establishing non-covalent interactions with the protein's surface, contribute to a remarkable stabilization of the enzyme. We used tetraethyl-orthosilicate (TEOS) and aminopropyl-triethoxysilane (APTES) as building blocks of the organosilica layer. Considering the dimension of the enzyme (7 nm), we used a 7-hour incubation time to fully cover (9 nm layer thickness) the enzyme inside the protection layer. For example, with this method, we could increase the half-life (𝑡50) of Biopox EV4 at 50°C from 18 minutes for the soluble enzyme to over 60 minutes for the protected enzyme. This will enable the enzyme to be active for a longer time in a harsh environment like high temperature used in the lamination process of multilayer packaging to degrade the adhesive polymer layer, which is the aim of the TERMINUS project.   Dataset This dataset contains all the UV-kinetic and immobilization endpoint raw data, SEM images, SDS-PAGE images, and Zeta potential to characterize and calculate the laccase immobilization, protection, and activity on silica nanoparticles used in Deliverable 1.5. Data are available in a compressed .zip file with one folder (Laccase-Biocatalytic-Redox-Systems_v1.0_TER_WP4_D4-1) containing seven files: Immobilization-BCA_D4-1-Laccase-Biocatalytic-Redox-Systems.xlsx contains all the BCA assay raw data for the concentration, immobilization, and immobilization optimization measurements. Kinetics_D4-1-Laccase-Biocatalytic-Redox-Systems.xlsx contains all the UV-kinetic activity assay raw data for soluble, immobilized, and protected enzymes in thermostability experiments. SDS-PAGE_D4-1-Laccase-Biocatalytic-Redox-Systems.pdf contains SDS-PAGE images obtained before and after dialysis for all the enzymes received from Biopox. SEM-images_D4-1-Laccase-Biocatalytic-Redox-Systems.pdf includes the SEM images used for characterization of silica nanoparticles before and after layer growth. Zeta-potential_D4-1-Laccase-Biocatalytic-Redox-Systems.pdf contains the raw data regarding the zeta potential measurements for silica nanoparticles and amino-modified silica nanoparticles. Materials_and_experimental_methods_D4-1-Laccase-Biocatalytic-Redox-Systems.pdf details the experimental method and conditions for the data acquisition presented in the data set. Guidance is also provided on using the data to calculate the laccase immobilization, activity, and thermostability. Metadata_information_D4-1-Laccase-Biocatalytic-Redox-Systems.pdf includes more detailed metadata information for the dataset represented in the excel files.