Difference between revisions of "Part:BBa K2271124"
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Valencene synthase converts FPP into Valencene. We chose the ValS from Callitropis nootkatensis, because other valencene synthases perform relatively inefficient in microorganisms. For example, they show a relatively poor product specificity, are less efficient in producing sesquiterpene than other synthases and have a significant amount of side products like germacrene A. In contrast to this, ValS from C. nootkatensis has a good product specificity and is more robust in terms of pH and temperature changes. It has also a better yield than citrus valencene synthases in yeast.
 
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This part additionally holds a PTS1 sequence for peroxisomal PEX5 import. For cytosolic expression please choose part: Bba_K2271118
 
  
 
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===Usage and Biology===
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===Usage and Biology===
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Valencene synthase converts FPP into Valencene. We chose the ValS from Callitropis nootkatensis, because other valencene synthases perform relatively inefficient in microorganisms[1]. For example, they show a relatively poor product specificity, are less efficient in producing sesquiterpene than other synthases and have a significant amount of side products like germacrene A. In contrast to this, ValS from C. nootkatensis has a good product specificity and is more robust in terms of pH and temperature changes. It has also a better yield than citrus valencene synthases in yeast.
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This part additionally holds a PTS1 sequence for peroxisomal PEX5 import. For cytosolic expression please choose part: Bba_K2271118<br>
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==Characterization==
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We verified the expression of ValS via western blot. Therefore, we have a 3xFLAG-6xHis-Tag as a part in our plasmids, we can use for the antibodies. The estimated atomic mass of ValS pts1 is 69 kDa.
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[[File:T--Cologne-Duesseldorf--western-blot-ValS.png|thumb|none|
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    <strong>Protein abundance in WT and transformed cells from <i>Saccharomyces cerevisiae</i>:</strong> Protein abundance was detected using 6x His Tag Antibody. WT = wild type, ValS = Valencene Synthase, PTS1 = Peroxisom Targeting Signal 1]]
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==References==
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[1] Jules Beekwilder et al. (2013) Valencene synthase from the heartwood of Nootka cypress (Callitropsis nootkatensis) for biotechnological production of valencene

Revision as of 18:40, 30 October 2017

ValS PTS1

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 860
    Illegal BamHI site found at 2732
    Illegal XhoI site found at 2768
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Usage and Biology

Valencene synthase converts FPP into Valencene. We chose the ValS from Callitropis nootkatensis, because other valencene synthases perform relatively inefficient in microorganisms[1]. For example, they show a relatively poor product specificity, are less efficient in producing sesquiterpene than other synthases and have a significant amount of side products like germacrene A. In contrast to this, ValS from C. nootkatensis has a good product specificity and is more robust in terms of pH and temperature changes. It has also a better yield than citrus valencene synthases in yeast.

This part additionally holds a PTS1 sequence for peroxisomal PEX5 import. For cytosolic expression please choose part: Bba_K2271118


Characterization

We verified the expression of ValS via western blot. Therefore, we have a 3xFLAG-6xHis-Tag as a part in our plasmids, we can use for the antibodies. The estimated atomic mass of ValS pts1 is 69 kDa.


Protein abundance in WT and transformed cells from Saccharomyces cerevisiae: Protein abundance was detected using 6x His Tag Antibody. WT = wild type, ValS = Valencene Synthase, PTS1 = Peroxisom Targeting Signal 1


References

[1] Jules Beekwilder et al. (2013) Valencene synthase from the heartwood of Nootka cypress (Callitropsis nootkatensis) for biotechnological production of valencene