Periodic Reporting for period 1 - FuturoLEAF (Leaf-inspired nanocellulose frameworks for next generation photosynthetic cell factories)
Reporting period: 2020-09-01 to 2021-08-31
FuturoLEAF will contribute significantly to society by developing the biotechnology field with direct impact to existing biotechnological companies utilizing photosynthetic cells. Additionally, it is envisioned that FuturoLEAF will enable the birth of new biotechnological companies, where solid-state cell factories will be utilized with one or multiple cell types, creating new business openings in the EU. The novel technology created within FuturoLEAF will also give EU a marketing advantage over other market areas related to bioproduction. Moreover, FuturoLEAF will contribute to environmental safety and the development of green technologies. Implementation of the knowledge created in FuturoLEAF for green technology will lead to sustainable use of resources by conversion of CO2 and light into chemicals.
Within the FuturoLEAF project, we have engineered recombinant green alga, C. reinhardtii, holding foreign cyclohexanone monooxygenase (CHMO) for converting cyclohexanone to the polymer precursor ε-caprolactone and optimized cultivation conditions for the highest conversion efficiency. We also showed an improved production of sucrose by sucrose-producing cyanobacterium Synechocystis sp. PCC 6803 entrapped in alginate beads.
In the production of monomers for biodegradable polymers, we have shown so far that the cells can be successfully immobilized in different materials and show excellent reaction rates. By molecular engineering, the photosynthetic chassis for monooxygenase reactions was optimized, leading to significantly elevated reaction rates. With the successful proof of concept both for the molecular engineering and implementation of biobased materials, we will focus on the performance of the system in 3D architectures, and the physiological characterization to match molecular cell engineering and material design in an iterative process.
In regard to the production of APIs, or more concretely the production of modified microcystins as novel payload class of antibody-drug conjugates (ADCs) for targeted cancer therapies, we could successfully demonstrate the proof of concept for the encapsulation of microcystin-producing cyanobacteria in both alginate and in nanocellulose.
The efforts towards the leaf-inspired hierarchical cell factory architecture with active immobilization strategy have been initiated. We have developed a FuturoLEAF -architecture allowing efficient light-capturing under low light conditions by creating a gradient of cells with truncated photosynthetic antennae across the immobilization matrix. This architecture can be further applied for improving production yields of commodity chemicals and biofuels, such as H2 and ethylene, as well as for improving the efficiency of biotransformation approaches.
In addition, molecularly imprinted polymers (MIPs) as synthetic receptors exhibiting molecular recognition of the substructures (peptide epitopes) of bacterial surface proteins have been developed, which allow specific binding to bacterial cells (cyanobacteria and E. coli). In addition, the interfacing of these MIPs with nanocellulose matrices has been studied. These combined findings and the resulting nanocomposite materials will allow the construction of self-assembling artificial leaf structures.
In regard to the production of active pharmaceutical ingredients (APIs), our preliminary results indicate that the production rates of modified microcystins in the reference system (alginate beads and in alginate films) are increased by almost two times compared to conventional suspension cultures.
The research related to active immobilization strategy involving MIPs and other approaches to reach the plant leaf-inspired architectures is expected to yield a generic approach to reversible and specific recognition and sequestering of target bacteria by stimuli-responsive nanocomposite materials. This will be an excellent example of biomimicry at the molecular level, based on the principles of bioinspiration and bioresources.
FuturoLEAF is expected to demonstrate a functional solid-state cell factory concept producing molecules for pharmaceuticals, commodity chemicals and fuels. FuturoLEAF will enable a significant step away from dependency of fossil sources, towards sustainable energy and chemicals production while contributing to the green production and climate change mitigation.