Part:BBa_K2271124

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

We used a valencene synthase(ValS) as part of our nootkatone pathway. ValS 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-PTS1 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