Difference between revisions of "Part:BBa K3195004"
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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. | 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. | ||
− | + | 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. | |
− | 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. | + | 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. |
− | [[Image:T--SEU-Nanjing-China--SLPdegraMCRR.png|760px|thumb|center|<b>Figure 1.</b>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)<b>The sample 12(in red) was degraded by subtilisin-like protease . | + | [[Image:T--SEU-Nanjing-China--SLPdegraMCRR.png|760px|thumb|center|<b>Figure 1.</b>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)<b>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. | 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 . | ||
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<span class='h3bb'>Sequence and Features</span> | <span class='h3bb'>Sequence and Features</span> |
Revision as of 01:19, 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.
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.
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
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Unknown
- 21INCOMPATIBLE WITH RFC[21]Unknown
- 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