Difference between revisions of "Part:BBa K1465205"
James zhang (Talk | contribs) (→iGEM2017 SZU-China) |
James zhang (Talk | contribs) (→iGEM2017 SZU-China) |
||
Line 37: | Line 37: | ||
The crude enzyme solution was obtained from the supernatant of fermentation broth of the Bacillus subtilis including the original strain WB800 and the recombinant WB800_ CsoS3. Since the fact that CA can also catalyze the hydration reaction of ester, which can be used to assay the activity of CA, we conducted measurement on CsoS3 according to a method from Verpoorte JA(1967), and the result(Fig.3) certified that the CsoS3 has high esterase activity. | The crude enzyme solution was obtained from the supernatant of fermentation broth of the Bacillus subtilis including the original strain WB800 and the recombinant WB800_ CsoS3. Since the fact that CA can also catalyze the hydration reaction of ester, which can be used to assay the activity of CA, we conducted measurement on CsoS3 according to a method from Verpoorte JA(1967), and the result(Fig.3) certified that the CsoS3 has high esterase activity. | ||
<div> | <div> | ||
− | <center><html><img src='https://static.igem.org/mediawiki/parts/2/26/CsoS3_3.png' style="width: | + | <center><html><img src='https://static.igem.org/mediawiki/parts/2/26/CsoS3_3.png' style="width:50%;margin:0 auto"> |
<center>Fig.3 The esterase activity of crude enzyme solution from the supernatant of fermentation broth of the Bacillus subtilis including the original strain WB800 and the recombinant WB800_ CsoS3.</center> | <center>Fig.3 The esterase activity of crude enzyme solution from the supernatant of fermentation broth of the Bacillus subtilis including the original strain WB800 and the recombinant WB800_ CsoS3.</center> | ||
</html></center> | </html></center> | ||
</div> | </div> |
Revision as of 18:12, 1 November 2017
Carbonic anhydrase (csoS3) of the carboxysome of Halothiobacillus neapolitanus
The carbonic anhydrase from Halothiobacillus neapolitanus converts incoming hydrogen carbonate into carbon dioxide inside the carboxysome. This step is essential for the CO2 fixation.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 166
- 1000COMPATIBLE WITH RFC[1000]
User Reviews
james_zhang
In 2017 SZU-China iGEM team changed the sequence of original part(BBa_K1465205) and the new part BBa_K2232022 was submited.
iGEM2017 SZU-China
We add the RBS of Bacillus subtilis and a segment of signal peptide at the upstream of cds of Carboxysomal Carbonic Anhydrase(csoS3) from Halothiobacillus neapolitanus, in order to achieve the expression and secretion of this part in the B.subtilis WB800. The construction of the shuttle expression-secretion vector_CsoS3 is shown as follow in figure 1.
The new part (BBa_K2232022) was constructed into shuttle expression-secretion vector pP43NMK, and 1% Agarose Gel Electrophoresis of the plasmid verified the correct construction in figure 2.
The crude enzyme solution was obtained from the supernatant of fermentation broth of the Bacillus subtilis including the original strain WB800 and the recombinant WB800_ CsoS3. Since the fact that CA can also catalyze the hydration reaction of ester, which can be used to assay the activity of CA, we conducted measurement on CsoS3 according to a method from Verpoorte JA(1967), and the result(Fig.3) certified that the CsoS3 has high esterase activity.