Part:BBa_K2934005
Signal peptide-protein linker for Bacillus subtilis
A short (9 amino acids) protein linker, used to connect a signal peptide to a secreted protein in Bacillus subtilis. This linker is derived from the commercial plasmid pBE-S (TaKaRa) that is designed to secrete recombinant proteins from Bacillus subtilis. The amino acid sequence is arg-pro-val-his-ile-trp-ser-ser-val.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Characterization
This sequence was used in our lab to connect a signal peptide (specifically AmyE) to a secreted enzyme (Glucose Oxidase and Invertase). It is one of the components in our composite parts- AmyE-Glucose Oxidase (2934004) and AmyE-Invertase (2934006). One of the important parameters of a linker is its ability to allow proper folding of the protein together with its conjugate.
In our experiments, we showed a successful secretion as well as extracellular activity of the Glucose Oxidase, with this linker at its N-terminus that connected between the enzyme and the signal peptide AmyE. Therefore, we assume that this sequence did not affect the properties of the protein.
However, we did not notice any band corresponding to the size of Invertase in SDS-PAGE (after concentrating the supernatant and lysate extracts), and the tested activity was present but very low. One possible explanation is that the peptide did interrupt the native folding and/or activity of this specific enzyme, although more explanations can be given for these results (low solution concentration, imprecise lab work, etc.).
For further reading regarding the results, please read the pages of AmyE-Glucose Oxidase (2934004), AmyE-Invertase (2934006) and lab results page on our wiki.
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