Part:BBa_K2940001

PelB and Bpul

The PelB leader sequence was joined to Bpul laccase to enable exportation of the enzyme into the periplasmic space. Both parts were isolated from the iGEM kit via Q5 PCR and joined together using Gibson Assembly. Not only was exporting Bpul outside of the cell vital for our overall project, it also allowed us to assess the activity of Bpul without the need for expensive and time consuming purification steps.

Usage and Biology

This biobrick was produced to be an integral part of our overall azo dye bioremediation strategy. The joining of the leader sequence PelB allows for the secretion of Bpul into the periplasmic space. PelB directs the unfolded fusion protein to the periplasm, where the peptide is cleaved by a signal peptidase. This causes the protein of interest (here Bpul) to be secreted into the periplasmic space.

Characterisation

To determine the effectiveness of the PelB leader sequence in secreting Bpul into the periplasmic and extracellular space SDS page was performed.