Difference between revisions of "Part:BBa K3887009"

Revision as of 21:52, 16 October 2021

Fre-L3-SttH

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal XhoI site found at 1813
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 942
Illegal NgoMIV site found at 1200
Illegal NgoMIV site found at 1903
Illegal NgoMIV site found at 2169
Illegal AgeI site found at 1088
Illegal AgeI site found at 1429
1000
INCOMPATIBLE WITH RFC[1000]
Illegal SapI.rc site found at 301
Illegal SapI.rc site found at 421

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 Fre-L3-SttH
-Fre-L3 is a type of flavin reductase (Fre). It is commonly used as a fusion enzyme. In our experiment, it is partnered with SttH. It can be highly expressed as a soluble form in E. coli and also regenerate FADH2 effectively. The expression SttH fusion enzyme is lower when using SttH alone, because it was affected by N-terminal domains. Fre was effective as a soluble N-terminal tag for SttH. Fre-L1–SttH, Fre-L2–SttH and Fre-L3– SttH are three fusion enzymes using three different linker sequence, resulting in different flexibility in fusing.
-During the experiment, ∆tnaA Fre-L3-SttH shows the highest concentration when conversion 6-Br-Trp from Trp. It is 39 times and 2.5 times higher than that of ΔtnaA SttH and ΔtnaA SttH+Fre, respectively. To test if other protein will increase the 6-Br-Trp production, Fre-L3 is compared with maltose-binding protein (MBP). MBP-L3-SttH also shows an increase in production of 6-Br-Trp. However, ∆tnaA MBP-L3-SttH shows 1.4 times increase compared to ∆tnaA SttH, while ∆tnaA Fre-L3-SttH produces 15-fold more. This experiment proves Fre efficiently enhanced SttH activity.
 <!-- Add more about the biology of this part here
 ===Usage and Biology===