Part:BBa_K4759204
T7-RBS4-SUMO-linker-OleP
We add a SUMO(solubilization-promoting tag) at the N-terminal of the P450 enzyme.
Usage and Biology
We add a solubilization-promoting tag at the N-terminal of the P450 enzyme, and in the selected six solubilization-promoting tags (SUMO, GST, MBP, TF, TRX, Nus), MBP and TF can significantly improve the solubility of Olep protein, but the enzyme activity of Olep is reduced, and the catalytic efficiency is lower than strain without solubilizing tag.
Small ubiquitin-related modifiers (SUMO) are extremely common in eukaryotes and are not found in prokaryotes. SUMO, commonly used as fusion tags, is derived from beer yeast with molecular weight 11.5kDa. The biggest characteristic of SUMO tag is that it can be specifically recognized by SUMO protease and highly degraded, making the subsequent tag removal process accurate and efficient. The SUMO tag can not only enhance the solubility of the recombinant protein, but also increase its expression level. The wild-type SUMO tag cannot be applied in a eukaryotic expression system because of the endogenous SUMO protease in eukaryotic cells. LifeSensors The company has developed an engineered SUMO tag and a supporting specific protease, enabling the SUMO tag to be applied to eukaryotic expression systems such as yeast, insect cells and mammalian cells.
Fig1: The application of protein tags Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 295
- 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 295
- 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 295
Illegal BglII site found at 184 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 295
- 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 295
Illegal AgeI site found at 420
Illegal AgeI site found at 518 - 1000COMPATIBLE WITH RFC[1000]
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