Chitin Deacetylase COD, BBa_K2848001
The BioBricks BBa_K2380044 and BBa_K2380043 (TU Darmstadt 2017), chitin deacetylase COD, were improved by adding a His6-tag to the C-terminus. As already proposed by the Darmstadt Team 2017 (ChiTUcare) this modification simplifies the purification of the protein by affinity chromatography. The resulting BioBricks are BBa_K2848002 and BBa_K2848001, respectively.
Additionally, the deacetylase COD (BBa_K2848002) was fused to the chitin synthase NodC (BBa_K2380002) to obtain BBa_K2848001. With this BioBrick E. coli should be capable of producing chitosan carrying only a single plasmid.
Pre-Nisin A, BBa_K2848004
The Biobrick BBa_K1365000 containing pre-Nisin A was improved during our project. Two distinctive tags were added to this BioBrick, resulting in BBa_K2848003. The first is a His6-tag which can be used for purification via immobilized metal affinity chromatography (IMAC). The second tag is the self-designed YAAY-tag consisting of Tyrosine-Alanine-Alanine-Tyrosine. Since nisin contains no tyrosines, this tag enables the link to other molecular compounds if a tyrosinase is used. Both tags are placed after the leader-peptide, which gets cleaved off after the processing of nisin. Thus, these two tags maintain their function even after processing of nisin and enable different downstream applications.