Coding

Part:BBa_K2271118

Designed by: Fiona Edenhofer   Group: iGEM17_Cologne-Duesseldorf   (2017-10-27)

ValS

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal suffix found in sequence at 2732
    Illegal BglII site found at 860
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Usage and Biology

You can use a valencene synthase(ValS) as part of the nootkatone pathway. ValS converts FPP into Valencene. You can 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 is designed for cytosolic expression, for peroxisomal import please choose part: BBa K2271124


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 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
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