Difference between revisions of "Part:BBa K3195004"

Revision as of 01:22, 16 October 2019

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 RR Degradation Efficiency

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.

Figure 1.The abscissa represent 28 enzymes in our project and the unit of ordinate value is mg/L. The ordinate value represents the remained microcystin RR(MC-RR) in the degraded sample. The left picture shows the results of degradation by each enzyme, and the right one shows the results of mixing each enzyme with enzyme 8(carboxypeptidase Z/N)The sample 12(in red) was degraded by subtilisin-like protease.

The results shows that subtilisin-like protease cannot degrade MC-RR effectively alone, but when mixed with carboxypeptidase Z/N, it can strongly degrade MC-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