Periodic Reporting for period 2 - ARTIBLED (Engineered ARTIficial proteins for Biological Light-Emitting Diodes)
Reporting period: 2021-01-01 to 2022-09-30
ARTIBLED proposes a three-step innovative approach to forge new artificial fluorescent proteins (AFPs) to enhance Bio-WLEDs meeting the technology needs for lighting. This project is realized by a highly interdisciplinary consortium comprising experts in Synthetic Biology, Organic Chemistry, Biocomputational, Theory, Photophysics, Hybrid Optoelectronics, and Industrial Bio-technology. CICbiomaGUNE will genetically modify protein scaffolds, while the UNITO will focus on the synthesis of new LED emitters. They are supported by TU Graz/CSIC for theoretical and computation simulations and TUM for spectroscopic measurements and coating fabrication. TUM will take over device fabrication and optimization, while ABIEL will focus on the upscale production of proteins. As such, the first year of ARTIBLED has been focused on designing suitable LED emitters and protein scaffolds using synthesis, spectroscopic and computational tools. Two family of emitters have been investigated achieving photoluminescence quantum yields >80% and >40 % for green and red emitters, respectively. The photo-induced degradation in polymer matrices is not fully understood yet. We have identified several suitable proteins and we are optimizing the preparation of the artificial fluorescent proteins. In the second period, UNITO/TUM/CSIC has explored up to 80 candidates, identifying 2 candidates for blue-green regions, 2 candidates for the yellow-emitting region and 1 candidate for the red-emitting region. This selection is based in terms of photoluminescence quantum yields and photostabilities. In parallel, CIC/TUGraz/CSIC have designed a robust computational approach to redefine the protein scaffolds to host a variety of LED emitters reaching high photostabilities and no loss of the photoluminescence quantum yield. This has been achieved for the red-emitting artificial fluorescent proteins that leads to devices meeting the expectations in terms of photostabilities (>1500 h). The relevance of this result is clearly stated by the direct comparison with devices prepared with the LED emitter ( 900 h stable) and the natural red-emitting protein (3 h). Finally, CSIC has elaborated a new computational protocol to study polymer coatings with proteins and UNITO/ABIEL have started with the up-scaling studies of LED emitters and protein scaffolds. Here, 98 % reduction cost of the protein scaffold has been realized.
As far as the scientific gains are concerned, ARTIBLED is focused on innovative designs of highly luminescence bio-hybrids using de novo proteins, modified LED emitters and new guidelines to replace natural emitters by new ones. Up to date, there is no rational design of proteins for tailored emitter suitable for lighting applications. This will be realized developing orthogonal aaRS/tRNA systems for bulky emitters at mg-scale using unnatural amino acid technology. Ideally, new guidelines will be developed to prepare novel artificial fluorescent proteins with remarkable photophysical features covering the visible range. The AFPs will be embedded into matrices to develop bio-phosphors towards the generation of new highly stable, low-cost, sustainable, and healthy bio-hybrid white light-emitting diodes operating under low and high powers. Up to date, bio-phosphors based on fluorescent proteins have been limited by the FP stability related to the chromophore deactivation under constant irradiation. Finally, our efforts will be completed by an efficient transfer of the laboratory protocols to industrial production (gr/day) redefining the recombinant protein strategies towards artificial fluorescent proteins. Thus, ARTIBLED is a unique opportunity to demonstrate the mature stage of Synthetic Biology towards providing a solution to a critical problem for the sustainability of WLEDs. In the second reporting period, we have optimized a red-emitting artificial fluorescent protein that is best performing than the LED emitter alone and the natural fluorescent proteins. The photostability, color and efficiency are already of interest for general lighting as well as for indoor farming, phototherapy, and high resolution microscopy.
As far as the scientific gains are concerned, ARTIBLED is focused on innovative designs of highly luminescence bio-hybrids using de novo proteins, modified LED emitters and new guidelines to replace natural emitters by new ones. Up to date, there is no rational design of proteins for tailored emitter suitable for lighting applications. This will be realized developing orthogonal aaRS/tRNA systems for bulky emitters at mg-scale using unnatural amino acid technology. Ideally, new guidelines will be developed to prepare novel artificial fluorescent proteins with remarkable photophysical features covering the visible range. The AFPs will be embedded into matrices to develop bio-phosphors towards the generation of new highly stable, low-cost, sustainable, and healthy bio-hybrid white light-emitting diodes operating under low and high powers. Up to date, bio-phosphors based on fluorescent proteins have been limited by the FP stability related to the chromophore deactivation under constant irradiation. Finally, our efforts will be completed by an efficient transfer of the laboratory protocols to industrial production (gr/day) redefining the recombinant protein strategies towards artificial fluorescent proteins. Thus, ARTIBLED is a unique opportunity to demonstrate the mature stage of Synthetic Biology towards providing a solution to a critical problem for the sustainability of WLEDs.