Part:BBa_K4188019

TvtGTPS1 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) to a dienol intermediate. This intermediate is reduced by the short chain dehydrogenase 1 (TvSDR1: BBa_K4188017) 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_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 919
12
INCOMPATIBLE WITH RFC[12]
Illegal PstI site found at 919
21
INCOMPATIBLE WITH RFC[21]
Illegal BglII site found at 1497
Illegal BamHI site found at 587
23
INCOMPATIBLE WITH RFC[23]
Illegal PstI site found at 919
25
INCOMPATIBLE WITH RFC[25]
Illegal PstI site found at 919
1000
COMPATIBLE WITH RFC[1000]