Part:BBa_K3855605
Sucrose-6-phosphat hydrolase sacA Expression casette for L. plantarum with an inducible Promotor and
Expression cassettes for the expression and secretion of Sucrose-6-phosphat hydrolase sacA in Lactobacillus. This enzyme facilitates the hydrolysis of terminal non-reducing beta-D-fructofuranoside residues in beta-D-fructofuranosides. It features an inducible promotor, which can be activated using Sakacin A Inducing Peptide/Sakacin P Inducing Peptide (SapIP/SppIP) (Sørvig et al. 2003 FEMS Microbiology Letters 229, 119-126).
This was necessary for us as we assembled the part using the type 2 s system in E.coli and the enzyme proved to be toxic to the host.
In the final host, Lactobacillus plantarum, we were able to induce the expression and demonstrate the secretion of the enzyme using a slot plot. This was possible as we added an N terminal his-tag to the enzyme.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal SpeI site found at 200
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 562
Illegal SpeI site found at 200 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1564
Illegal BamHI site found at 1245 - 23INCOMPATIBLE WITH RFC[23]Illegal SpeI site found at 200
- 25INCOMPATIBLE WITH RFC[25]Illegal SpeI site found at 200
- 1000COMPATIBLE WITH RFC[1000]
| None |