NUMAFERM’s platform technology
Available bioprocesses are successfully applied for the production of many enzymes (and other proteins). Batch sizes in the tons scale are standard and the production costs can be as low as 10-20 €/kg (van Beilen et al., Current opinion of biotechnology, 2002). For the production of peptides, however, available processes are limited by three major bottlenecks: 1. omnipresence of proteases destroying peptides (figure 1), 2. aggregation of peptides complicating downstreaming processes (DSP) and 3. toxic effects of peptides towards the production host.
NUMAFERM’s technology copes with these challenges and serves as a reliable and cost-efficient platform. How have we achieved this success? We have developed the first-in-place secretion technology for Gram-negative bacteria; our working horses are common E. coli strains. Peptides are transported out of the cells into to the protease-free surrounding immediately after biosynthesis. This protects them from being destroyed (figure 1 and 2). Our technology is based on a Type 1 secretion system (T1SS) where a transport complex of three membrane proteins catalyzes the transport. Requested peptides are genetically fused with a transport signal that is recognized by the nanomachinery (figure 2). After transport, the transport signal remains attached to the peptide, which is essential. First, we rationally designed the transport signal in such a way that it increases the stability and solubility of peptides and counteracts aggregation. Second, the return of peptides to the production host is prevented by the transport signal – cells are protected from cytotoxic effects.
Fusion peptides are highly pure (figure 1) and can be easily extracted from the culture supernatant. The transport signal is cleaved-off during the DSP and pure peptide is produced.
In this way NUMAFERM’s technology solves limitations of available technologies and enables the cost-efficient production of peptides.