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Biology subjects

Timonen, J. V. I.

Publications and source records attributed to Timonen, J. V. I..

2 recordsLinked to original sources

Supercharged Fluorescent Protein-Apoferritin Cocrystals for Lighting Applications

The design of lighting sources based on fluorescent proteins (FPs) has been limited by the lack of protocols to stabilize FPs under preparation (deposition techniques, organic solvents, etc.) and working (temperature, irradiation, etc.) conditions. As a critical bottleneck, photo-induced heat generation due to FP motion and quick heat transfer leads to working device temperatures of ca. 70 {degrees}C, resulting in a quick FP-denaturation and, in turn, a quick loss of the device performance. Herein, we showcase FP stabilization for lighting devices with an electrostatically self-assembled FP-apoferritin cocrystals embedded in a silicone-based color down-converting filter. This strategy highlights three major advances: i) engineering of positively supercharged FPs (+22) without losing photoluminescence and thermal stability compared to its native form, ii) a crystallization protocol resulting in highly emissive cocrystals keeping the photoluminescence features of the FPs, and iii) a 40-fold increase of the lighting device stability compared to reference devices due to the reduction of the device working temperatures to 40 {degrees}C. Thus, the success of this multidisciplinary approach contributes toward developing stable energy-related protein-based optoelectronics.

bioengineering↗

Recombinant protein condensation inside E. coli enables the development of building blocks for bioinspired materials engineering - biomimetic spider silk protein as a case study

Recombinant expression of proteins destined to form biological materials often results in poor production yields or loss of their function due to premature aggregation. Recently, liquid-liquid phase separation has been proposed as a mechanism to control protein solubility during expression and accumulation in the cytoplasm. Here, we investigate this process in vivo during the recombinant overexpression of the spider silk-mimetic mini-spidroin NT2RepCT in Escherichia coli. The protein forms intracellular liquid-like condensates that shift to a solid-like state triggered by a decrease in their microenvironmental pH. These features are also maintained in the purified sample in vitro both in the presence of a molecular crowding agent mimicking the bacterial intracellular environment, and during a biomimetic extrusion process leading to fiber formation. Overall, we demonstrate that characterization of protein condensates inside E. coli could be used as a basis for selecting proteins for both materials applications and their fundamental structure-function studies.

synthetic biology↗