Difference between revisions of "Part:BBa K3195004"

Revision as of 01:07, 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.

Usage and Biology

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. Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal BglII site found at 447
Illegal XhoI site found at 1645
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal AgeI site found at 889
Illegal AgeI site found at 1120
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI.rc site found at 967

@@ Line 6: / Line 6: @@
 ===Usage and Biology===
-Similar to subtilisin, our subtilisin-like protease can catalyze the hydrolysis of protein to amino acid. In industry subtilisin has been widely used in all walks of life. 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.
+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.
-[[Image:T--SEU-Nanjing-China--SLPdegraMCRR.png|560px|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 in the degraded sample. The left picture shows the results of degradation by each enzyme, and the right one shows the results of mix each enzyme with enzyme 8(carboxypeptidase Z/N)<b>The sample 12(in red) was degraded by subtilisin-like protease .</b>]]
+[[Image:T--SEU-Nanjing-China--SLPdegraMCRR.png|560px|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 .</b>]]
+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.
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 <span class='h3bb'>Sequence and Features</span>