Spin-Off project SenseUp
Heads of SenseUp-Team
Dr. Stephan Binder
Institute of Bio- and Geosciences
Forschungszentrum Jülich GmbH
- Video Innovationspreis NRW
- Video clip, Science4Life Konzeptphase 2014 (Position 7m47s), Die 20 Nominierungen
- Abstract GO-Bio, Schnellere Stammentwicklung für die industrielle Biotechnologie
- General Information, Gründungsoffensive Biotechnologie GO-Bio
- Gründerportrait der Helmholtzgemeinschaft, www.helmholtz.de/technologie/mit-leuchtenden-bakterien-zum-eigenen-unternehmen-4765
Highly efficient microbial producer strains are an essential component of many biotechnological processes. While there is an increasing need for novel and enhanced producer strains, their development and construction is limited by laborious and time-consuming techniques.
Major bottlenecks in strain development are screening steps, in which millions of genetic variants have to be analyzed for enhanced productivity under defined process-conditions. With current techniques, this often equals the proverbial search for a needle in a haystack. This results in significant limitations in strain development:
- Limited screening-capacity denies access to full genetic diversity and thus limits development potential
- Limited screening-capacity makes complex genome engineering very laborious or even impossible
- Individual cultivation of every clone restricts screening cultures to flask- or MTP-format, thereby ignoring process-relevant culture conditions (e.g. Fed-batch, pH-control, aeration,…).
Finding the needle in the haystack
The SenseUp Technology overcomes these problems, enabling integrated strain and process-development in ultra-high throughput (uHT).
It is based on intracellular genetic sensors, which transform product concentrations into a graded optical output (Fig. 1). In combination with fluorescence activated cell sorting (FACS) this enables product-oriented screenings of up to 50,000 individual cells per second and direct isolation of the best producing cells. This corresponds to million-fold acceleration of the screening process and generates significant advantages of the SenseUp-Technology compared to conventional techniques (Fig. 2):
- Full diversity-coverage, accessing full development potential
- Complex genome engineering, enabling tailor-made development
- Integrated process development, resulting in ready to use strains
Creating a technology platform
The SenseUp technology is established for the detection of various amino acids, precursor metabolites and NADPH in the industrially relevant hosts Corynebacterium glutamicum and Escherichia coli. Several applications were successfully demonstrated, for example, screening of:
- large mutant cell libraries, obtained by undirected genome-wide mutagenesis for enhanced lysine production1.
- gene-specific libraries, derived from error-prone PCR, for feedback-resistant variants of key enzymes for lysine, arginine, and histidine biosynthesis2.
- saturation libraries of selected codons in essential target genes for alleles with a positive influence on productivity2.
- gene-specific libraries of NADPH-dependent dehydrogenases for variants with altered substrate specificity3.
The SenseUp-project was initiated in 2014 with financial support of the Helmholtz Enterprise Fonds and Forschungszentrum Jülich. After being successful in the GO-Bio initiative (Gründungsoffensive Biotechnologie), the project will be supported by the BMBF until 2017.
Our aim is to expand the SenseUp-technology platform and to bring it to market maturity in the end of 2016. SenseUp will offer tailor-made microbial production processes for amino acids and customized technical enzymes.
- Binder, S. et al. A high-throughput approach to identify genomic variants of bacterial metabolite producers at the single-cell level Genome Biol 13, R40 (2012).
- Schendzielorz, G. et al. Taking Control over Control: Use of Product Sensing in Single Cells to Remove Flux Control at Key Enzymes in Biosynthesis Pathways. ACS Synth. Biol. 3, 21–29 (2014).
- Siedler, S. et al. SoxR as a Single-Cell Biosensor for NADPH-Consuming Enzymes in Escherichia coli. ACS Synth. Biol. 3, 41–4 (2014).
- Binder, S. et al. Recombineering in Corynebacterium glutamicum combined with optical nanosensors: a general strategy for fast producer strain generation. Nucleic Acids Research 41, 6360–6369 (2013).