Difference between revisions of "Part:BBa K2117012"
Line 3: | Line 3: | ||
<partinfo>BBa_K2117012 short</partinfo> | <partinfo>BBa_K2117012 short</partinfo> | ||
− | + | 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. | |
− | |||
===Usage and Biology=== | ===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. | ||
+ | |||
+ | |||
<!-- --> | <!-- --> |
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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE 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 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]