Genetically encoded FRET-based biosensors are a cutting-edge technology to measure metabolite concentrations inside living cells. The aim of this project is to unlock the potential of extracellular FRET sensors for the development of biorefinery processes. We plan the development of a FRET-based xylose sensor toolbox, which enables the online detection of xylose as an alternative C-source in mini pilot plants. The project is linked to the FocusLab HyImpact and can also contribute valuable results to AP3. In both FocusLabs 2nd generation raw material from renewable resources is used to produce xylose from hemicellulose. The use of xylose in addition to glucose is mandatory to develop economical biomass-based processes. The xylose sensors, which are developed in XyloSenS are essential for the high-throughput process development specifically of respective microbial strains accepting this pentose sugar as a C-source and can also be used for optical process control. In XyloSenS we will develop a toolbox of xylose sensors with different affinities to cover a broad range of xylose concentrations.
The transformation of a petrol-based chemistry to renewable resources is one of the biggest tasks for the future. In order to avoid competition with crops for human and animal nutrition, the use of waste streams is decisive for a circular economy. Biogenic waste streams can contribute to the economy and efficiency of novel biotechnological processes by delivering amongst others valuable carbohydrates as a C-source for microbial biotransformations. To make use of 2nd generation raw material as a C-source for microbial transformations, a sensor detecting the predominant sugar D-xylose from respective hydrolysates (bagasse, lignocellulose) is extremely useful, not only for the development of respective microbial strains but also for the control of such processes by making use of the fluorescence signal delivered by such sensors. XyloSenS is directly linked to the FocusLab HyImPAct, which uses Corynebacterium glutamicum as a platform organism. In order to generate respective C. glutamicum strains for the utilization of D-xylose containing feedstocks with minimized carbon loss, the novel xylose sensors will be applied as an online screening tool. Furthermore, such sensors are also useful to control the respective biotransformations carried out by using such optimized strains. XyloSenS contributes directly to the profiling of BioSC by the development of a basic analytic technology relevant for the fast and efficient development of biorefineries. The here described sensor development is not only suitable for the detection of sugars, but can also be applied for the measurement of other metabolites, such as metabolites produced by microbial sources. There is a general need for speed in microbial strain development. Parallelization in small scale is often limited by analysis of target compounds in the cultivation devices. The application of FRET-based biosensors is a promising technology to tackle the problem. The online fluorescence signal delivered by FRET-based biosensors in the 48-well format of a microtiter plate used in a BioLector® enables direct analysis of the 48 parallel cultivations. Currently, off-line analysis by HPLC is necessary to determine the content of different sugars in the cultivation broth. This offline-analytics represents a bottle neck for parallelized strain development. It can be expected that the time for strain development will be halved by the application of the FRET-based sensors compared to the common practice. To the best of our knowledge such biosensors are up to now almost exclusively applied in vivo by other groups. Thus, our approach can be regarded as unique.
Prof. Martina Pohl
Forschungszentrum Jülich GmbH (FZJ)
phone: +49 2461 614388
Prof. Dr. Wolfgang Wichert & Prof. Dr. Martina Pohl, IBG-1, Forschungszentrum Jülich GmbH
Prof. Dr. Dieter Willbold & Dr. Bernd König, Biophysics of Protein Interactions, Heinrich Heine University Düsseldorf
Prof. Dr. Jochen Büchs, Biochemical Engineering, RWTH Aachen University
01.10.2018 - 30.09.2020
The total budget of XyloSenS is 155,886 €. XyloSenS is is part of the NRW-Strategieprojekt BioSC and thus funded by the Ministry of Culture and Science of the German State of North Rhine-Westphalia.