Difference between revisions of "Part:BBa K2560262"
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<partinfo>BBa_K2560262 short</partinfo> | <partinfo>BBa_K2560262 short</partinfo> | ||
− | + | 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.
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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 885
Illegal BamHI site found at 213 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 1188
Illegal AgeI site found at 1270 - 1000COMPATIBLE WITH RFC[1000]