Line 10: | Line 10: | ||
<p>When it comes to choosing a production host, Bacillus subtilis is particularly interesting: the Gram-positive model organism can be easily genetically modified and has powerful secretion capacities. <a target="_blank" href ="https://www.researchgate.net/profile/Reindert_Nijland/publication/23656731_Optimization_of_Protein_Secretion_by_Bacillus_subtilis/links/09e415136142e1554c000000/Optimization-of-Protein-Secretion-by-Bacillus-subtilis.pdf">[1]</a></p> | <p>When it comes to choosing a production host, Bacillus subtilis is particularly interesting: the Gram-positive model organism can be easily genetically modified and has powerful secretion capacities. <a target="_blank" href ="https://www.researchgate.net/profile/Reindert_Nijland/publication/23656731_Optimization_of_Protein_Secretion_by_Bacillus_subtilis/links/09e415136142e1554c000000/Optimization-of-Protein-Secretion-by-Bacillus-subtilis.pdf">[1]</a></p> | ||
<p>In this part of EncaBcillus we aimed at making use of the B. subtilis native advantages and combined them with Peptdiosomes – a new innovative cultivation platform for functional co-cultivation. Multiple Peptidosomes, with each encapsulating one specific strain that secrets one protein of interest. By doing so, the production of multi-complex proteins cloud be achieved by separated subpopulations all in one reaction hub.</p> | <p>In this part of EncaBcillus we aimed at making use of the B. subtilis native advantages and combined them with Peptdiosomes – a new innovative cultivation platform for functional co-cultivation. Multiple Peptidosomes, with each encapsulating one specific strain that secrets one protein of interest. By doing so, the production of multi-complex proteins cloud be achieved by separated subpopulations all in one reaction hub.</p> | ||
− | <p>To ensure the assembly of proteins outside of the Peptidosomes we further characterized the SpyTag/SpyCatcher system. Theses functional units can be attached to any protein of interest and upon secretion will result in a covalent isopeptide bond between the SpyTag/SpyCatcher partners. | + | <p>To ensure the assembly of proteins outside of the Peptidosomes we further characterized the SpyTag/SpyCatcher system. Theses functional units can be attached to any protein of interest and upon secretion will result in a covalent isopeptide bond between the SpyTag/SpyCatcher partners <a target="_blank" href =" https://www.ncbi.nlm.nih.gov/pubmed/28230977">[2]</a>. |
The system originates from Streptococcus pyogenes and since it’s discovery it has been under constant development (3). In our project we applied codon-adapted B. subtilis specific tags and reduced the SpyCatcher in length to enhance it´s usability when translationally fused to a protein of interest. Thus, decreasing the chances of the tag interfering with overall protein folding. (4)</p> | The system originates from Streptococcus pyogenes and since it’s discovery it has been under constant development (3). In our project we applied codon-adapted B. subtilis specific tags and reduced the SpyCatcher in length to enhance it´s usability when translationally fused to a protein of interest. Thus, decreasing the chances of the tag interfering with overall protein folding. (4)</p> | ||
<p>To demonstrate the applicability of both tags we fused them to a green (sfGFP) and a red (mCherry) fluorescent protein, enabling an easy detectable output. (For more details please check our Design section) | <p>To demonstrate the applicability of both tags we fused them to a green (sfGFP) and a red (mCherry) fluorescent protein, enabling an easy detectable output. (For more details please check our Design section) |
Revision as of 14:42, 26 October 2017