Difference between revisions of "Part:BBa K2560261"

Revision as of 13:24, 30 September 2018

mcrC - C-terminal domain of Malonyl-CoA Reductase from Chloroflexus aurantiacus

This part contains the 3' 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-hydroxypropionic 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 BBa_K2560261 and this part. If the gene for the whole enzyme shall be used take BBa_K2560260.

For this part, mcrC 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
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 288
Illegal NheI site found at 1690
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 361
Illegal NgoMIV site found at 1975
Illegal AgeI site found at 1524
1000
COMPATIBLE WITH RFC[1000]

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K2560261 short</partinfo>
-This part contains the 3' 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-hydroxypropionic 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.
+This part contains the 3' 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-hydroxypropionic 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.
 [[File:BBa K2560260 Mcr pathway.png|500px| center]]
-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 [https://parts.igem.org/wiki/index.php?title=Part:BBa_K2560261 BBa_K2560261] and this part. If the gene for the whole enzyme shall be used take [https://parts.igem.org/wiki/index.php?title=Part:BBa_K2560260 BBa_K2560260]
+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 [https://parts.igem.org/wiki/index.php?title=Part:BBa_K2560261 BBa_K2560261] and this part. If the gene for the whole enzyme shall be used take [https://parts.igem.org/wiki/index.php?title=Part:BBa_K2560260 BBa_K2560260].
 For this part, <i>mcrC</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).