Coding

Part:BBa_K4188017

Designed by: Tobias Bröker   Group: iGEM22_WWU_Muenster   (2022-09-30)


TvSDR1 fused to PTS1 tag for the peroxisomal expression

In Thymus vulgaris, GPP is converted to γ-terpinene by the γ-terpinene synthase 1 (TvGTPS1: BBa_K4188028 + PTS1, BBa_K4188019), it is oxidized by a cytochrom P450 monooxygenase (CYP) of the 71D family (TvCYP71D179: BBa_K4188018 + PTS1, BBa_K4188030) to a dienol intermediate. This intermediate is reduced by the short chain dehydrogenase 1 (TvSDR1: BBa_K4188017 + PTS1, BBa_K4188029) to an instable allylic ketone intermediate which rearranges to thymol (Krause et al. 2021). The gene is MoClo (RFC1000) compatible.


Figure 1: Biosynthesis of thymol. Geranyl pyrophosphate (GPP) is converted to γ-terpinen by the Thymus vulgaris γ-terpinen-synthase 1 (TvGTPS1). Further conversion to a dienol intermediate is mediated by thymes CYP71D179. The T. vulgaris short chain dehydrogenase (TvSDR1) converts the dienol intermediate to an instable allylic ketone which rearranges to thymol.


Genes for the thymol production

part number Gene length
BBa_K4188017 TvSDR1_PTS1 804 bp
BBa_K4188018 TvCYP71D179_PTS1 1500 bp
BBa_K4188019 TvtGTPS1_PTS1 1638 bp
BBa_K4188029 TvSDR1 795 bp
BBa_K4188030 TvCYP71D179 1491 bp
BBa_K4188028 TvtGTPS1 1629 bp

Reference

Krause, S. T., Liao, P., Crocoll, C., It Boachon, B., F€ Orster, C., Leidecker, F., Wiese, N., Zhao, D., Wood, J. C., Buell, C. R., Gershenzon, J., Dudareva, N., & Org Degenhardt, J. (n.d.). The biosynthesis of thymol, carvacrol, and thymohydroquinone in Lamiaceae proceeds via cytochrome P450s and a short-chain dehydrogenase. https://doi.org/10.1073/pnas.2110092118/-/DCSupplemental

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 529
    Illegal PstI site found at 766
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 529
    Illegal PstI site found at 766
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 529
    Illegal PstI site found at 766
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 529
    Illegal PstI site found at 766
    Illegal NgoMIV site found at 164
    Illegal NgoMIV site found at 277
    Illegal NgoMIV site found at 289
    Illegal NgoMIV site found at 460
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None