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Risultati finali
A functional Twin-Arginine Translocation pathway (TAT) was demonstrated in Bacillus subtilis and Escherichia coli. These organisms are the models for, respectively, Gram-positive and Gram-negative bacteria. In E. coli, the TatA, TatB, and TatC proteins are the main components of the translocation machinery. In B.subtilis, TatB is absent, but TatA and TaC are present in two pairs.
These pairs have a strong substrate specificity: Tat(A/C)d is required for the transport of the phosphodiesterase, PhoD, whereas Tat(A/C)y is required for YwbN transport.
In E.coli, several combinations of TAT-specific signal peptides with cytoplasmic and extracellular proteins were actively transported across the Inner membrane. In one case, this concerned the Green-Fluorescent Protein (GFP), which was clearly transported in a folded state. Overproduction of Tat components in both B. subtilis and E.coli showed can be achieved. This system has the potential to be used for the production of native and heterologus secreted proteins, which are not (efficiently) secreted via the major, Sec, pathway for protein secretion.
These findings were patented by the partner company, Genencor International.
Further information on the project can be found at: http://www.ncl.ac.uk/bacell/
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