Difference between revisions of "Part:BBa K3722004"

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===Biology===
 
===Biology===
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Two flexible linkers and one rigid linker were compared, considering that the linker may affect substrate diffusion to the active site pocket or the flexibility of such linkers may reduce activity[1]. Finally found that the chimeric enzymes showed highly increased ratios of their soluble forms in E. coli. The molar concentrations of soluble Fre-Linker–SttH was 1.64 μM, which was more than 3 times higher than that of SttH by itself[2].
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It is a rigid peptide, which is used to completely isolate the two connected proteins and maintain their independent functions, because some flexible peptides will cause the expression of the fusion protein to decrease or the activity to be lost. It should be noted that rigid peptides cannot be used for two interacting proteins.
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        <img src="https://2021.igem.org/wiki/images/e/ee/T--NWU-CHINA-A--part04.png" width="80%" style="float:center">
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===Usage===
 
===Usage===
 
The part was used to construct SttH and Fre fusion enzyme to enhance the solubility and activity of SttH. Due to the limited time, we did not characterize this part. As a result, there is a lack of data about this part.
 
The part was used to construct SttH and Fre fusion enzyme to enhance the solubility and activity of SttH. Due to the limited time, we did not characterize this part. As a result, there is a lack of data about this part.
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<html>
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    <figure>
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        <img src="https://2021.igem.org/wiki/images/2/2b/T--NWU-CHINA-A--part01.png" width="100%" style="float:center">
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        <figcaption>
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        <p style="font-size:1rem">
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        </p>
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        </figcaption>
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    </figure>
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===References===
 
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[1] Aalbers, F . S. & Fraaije, M. W . Coupled reactions by coupled enzymes: alcohol to lactone cascade with alcohol dehydrogenase-cyclohexanone monooxygenase fusions. Appl. Microbiol. Biotechnol. 101, 7557–7565 (2017).
 
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[2] Lee, J., Kim, J., Song, J.E. et al. Production of Tyrian purple indigoid dye from tryptophan in Escherichia coli. Nat Chem Biol 17, 104–112 (2021). https://doi.org/10.1038/s41589-020-00684-4
 
  
  

Latest revision as of 21:17, 21 October 2021


A linker sequence in fusion protein Fre-Linker-SttH

A linker sequence belongs to our fusion protein Fre-Linker-SttH, coding a rigid linker between Fre and SttH.


Biology

It is a rigid peptide, which is used to completely isolate the two connected proteins and maintain their independent functions, because some flexible peptides will cause the expression of the fusion protein to decrease or the activity to be lost. It should be noted that rigid peptides cannot be used for two interacting proteins.


Usage

The part was used to construct SttH and Fre fusion enzyme to enhance the solubility and activity of SttH. Due to the limited time, we did not characterize this part. As a result, there is a lack of data about this part.




Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]