Part:BBa_K3195004
Trx-His-EK tagged subtilisin-like protease
Subtilisin-like protease is a kind of alcalase found in amphioxus (Branchiostoma) similar to subtilisin protease. In our project, it has been used to degrade microcystin with another enzyme called carboxypeptidase Z/N.
Microcystin Degradation Efficency
Similar to subtilisin, our subtilisin-like protease can catalyze the hydrolysis of protein to amino acid. Subtilisin has been widely used in all walks of life even in laundry. Interestingly, we found the subtilisin-like protease can degrade not only proteins but also microcystins. Then we test the activity of this enzyme and measured by HPLC-MS-MS.
In the wet lab experiments, we found that carboxypeptidase Z/N and subtilisin-like protein could not decompose microcystins independently, but when the two enzymes were put into the experiment, they could decompose one of the microcystins, microcystin RR.
In our project, we cloned this part into pET32a vector and transformed it into E.coli(DH5α) for mass production of subtilisin-like protease .
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 447
Illegal XhoI site found at 1645 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 889
Illegal AgeI site found at 1120 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 967
//chassis/prokaryote/ecoli
//function/degradation
origin | Branchiostoma |
protein | subtilisin-like protease |
tag | Trx-His-EK |