Difference between revisions of "Part:BBa K2560262"

 
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<partinfo>BBa_K2560262 short</partinfo>
 
<partinfo>BBa_K2560262 short</partinfo>
  
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This part contains the 5' part of the <i>mcr</i> gene for Malony-CoA Reductase from <i>C. aurantiacus</i>, encoding the C-terminal domain. This enzyme is involved in the 3-hydroxypropionate cycle for CO2 fixation and converts malonyl-CoA into 3-hydroxy propionic acid (Strauss & Fuchs, 1993). In a two-step reaction malonyl-CoA is first reduced to 3-oxopropanoic acid and then further reduced to 3-hydroxypropionic acid. The C-terminal domain catalyzes the first reaction step to 3-oxopropanoic acid and the N-terminal domain catalyzes the second reaction step to 3-hydroxypropionic acid.
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[[File:BBa K2560260 Mcr pathway.png|500px| center]]
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It was shown that a splitted version of Mcr with separated C- and N-terminal domains increases enzyme activity (Liu et al., 2013). If these splitted genes shall be used take this part and [https://parts.igem.org/wiki/index.php?title=Part:BBa_K2560261 BBa_K2560261]. If the gene for the whole enzyme shall be used take [https://parts.igem.org/wiki/index.php?title=Part:BBa_K2560260 BBa_K2560260].
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For this part, <i>mcrN</i> was codonoptimized for <i>V. natriegens</i> and then synthetisized and integrated into the vector [https://parts.igem.org/Part:BBa_K2560002 BBa_K2560002] via BsmBI. According to Liu et al, we designed the part so that McrN contains amino acid 1-549 of the overall protein and McrC amino acid 550-1219 (Liu et al., 2013).
  
 
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Revision as of 14:12, 30 September 2018


mcrN - N-terminal domain of Malonyl-CoA Reductase from Chloroflexus aurantiacus

This part contains the 5' part of the mcr gene for Malony-CoA Reductase from C. aurantiacus, encoding the C-terminal domain. This enzyme is involved in the 3-hydroxypropionate cycle for CO2 fixation and converts malonyl-CoA into 3-hydroxy propionic acid (Strauss & Fuchs, 1993). In a two-step reaction malonyl-CoA is first reduced to 3-oxopropanoic acid and then further reduced to 3-hydroxypropionic acid. The C-terminal domain catalyzes the first reaction step to 3-oxopropanoic acid and the N-terminal domain catalyzes the second reaction step to 3-hydroxypropionic acid.

BBa K2560260 Mcr pathway.png

It was shown that a splitted version of Mcr with separated C- and N-terminal domains increases enzyme activity (Liu et al., 2013). If these splitted genes shall be used take this part and BBa_K2560261. If the gene for the whole enzyme shall be used take BBa_K2560260.

For this part, mcrN was codonoptimized for V. natriegens and then synthetisized and integrated into the vector BBa_K2560002 via BsmBI. According to Liu et al, we designed the part so that McrN contains amino acid 1-549 of the overall protein and McrC amino acid 550-1219 (Liu et al., 2013).

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 885
    Illegal BamHI site found at 213
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1188
    Illegal AgeI site found at 1270
  • 1000
    COMPATIBLE WITH RFC[1000]