Difference between revisions of "Part:BBa K2117012"

Revision as of 18:03, 19 October 2016

CrtE improved

Improved version of biobrick encoding the CrtE enzymes, which catalyses a step in the beta-carotene pathway. Yarrowia lipolytica ribosome binding site – CACA, was added in front of the ORF and removing an illegal restriction site making the Biobrick compatible with the RFC25 standard for in-frame protein fusion.

Usage and Biology

Beta-carotene is naturally produced by a range of organisms such as plants and fungi, but neither conventional yeast nor Y. lipolytica has the pathway for biosynthesis. Beta-carotene is produced by four enzymatic steps from farnesyl diphosphate (F-PP), which is naturally produced in Y. lipolytica. In the next step, farnesyl diphosphate is converted to geranylgeranyl diphosphate (GG-PP) in a reaction catalyzed by geranylgeranyl diphosphate synthase (CrtE). GG-PP is transformed to phytoene by CrtYB, which is an enzyme with two domains, one functioning as phytoene synthase and another as lycopene cyclase, in this reaction the first domain plays a crucial role. The next step results in production of lycopene and is catalyzed by carotene desaturase (CrtI). Finally, lycopene is converted by CrtYB with the lycopene cyclase domain into beta-carotene6.

The JHU 2011 iGEM team produced beta-carotene in Saccharomyces cerevisiae by constructing three biobricks with the three individual genes encoding the enzymes from the pathway from the fungi Xanthophyllomyces dendrorhous.

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 335
Illegal XhoI site found at 34
Illegal XhoI site found at 166
Illegal XhoI site found at 985
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K2117012 short</partinfo>
-Will come later
+Improved version of biobrick encoding the CrtE enzymes, which catalyses a step in the beta-carotene pathway. <i>Yarrowia lipolytica</i> ribosome binding site – CACA, was added in front of the ORF and removing an illegal restriction site making the Biobrick compatible with the RFC25 standard for in-frame protein fusion.
-<!-- Add more about the biology of this part here
 ===Usage and Biology===
+Beta-carotene is naturally produced by a range of organisms such as plants and fungi, but neither conventional yeast nor Y. lipolytica has the pathway for biosynthesis. Beta-carotene is produced by four enzymatic steps from farnesyl diphosphate (F-PP), which is naturally produced in Y. lipolytica. In the next step, farnesyl diphosphate is converted to geranylgeranyl diphosphate (GG-PP) in a reaction catalyzed by geranylgeranyl diphosphate synthase (CrtE). GG-PP is transformed to phytoene by CrtYB, which is an enzyme with two domains, one functioning as phytoene synthase and another as lycopene cyclase, in this reaction the first domain plays a crucial role. The next step results in production of lycopene and is catalyzed by carotene desaturase (CrtI). Finally, lycopene is converted by CrtYB with the lycopene cyclase domain into beta-carotene6.
+The JHU 2011 iGEM team produced beta-carotene in <i>Saccharomyces cerevisiae</i> by constructing three biobricks with the three individual genes encoding the enzymes from the pathway from the fungi Xanthophyllomyces dendrorhous.
 <!-- -->