Part:BBa_K3140000
PsiD - Tryptophan decarboxylase from Psilocybe cubensis
PsiD is a tryptophan decarboxylase that catalyses the conversion of L-tryptophan to tryptamine.
- NCBI: ASU62239.1
- UniProt: P0DPA6
- EC number: 4.1.1.105
Usage and Biology
The mechanism of psilocybin biosynthesis in Psilocybe sp. was recently elucidated by Fricke et al. (2017). L-tryptophan proceeds through decarboxylation (mediated by PsiD), hydroxylation (mediated by PsiH), phosphorylation (mediated by PsiK), and finally N,N-dimethylation (mediated by PsiM) to yield psilocybin.
PsiD is a native enzyme obtained from the Psilocybe cubensis, which is involved in the metabolic biosynthesis of psilocybin from tryptophan. It accepts both L-tryptophan and 4-hydroxy-L-tryptophan as substrates, producing tryptamine (Fig. 1) and 4-hydroxytryptamine (Fig. 2), respectively. In a native state, PsiD is a 439 amino acid protein (49.6 kDa) with a theoretical pI of 5.44 calculated with the ExPASy ProtParam tool.
Heterologous expression of PsiD has been achieved in a T7 induction system using pET-28c(+) transformed into Escherichia coli BL21(DE3), co-transformed with chaperone plasmid pGro7 (Fig. 3), resulting in a 475 amino acid polypeptide, with a computed molecular weight of 53.6 kDa.
A band consistent with expression of PsiD in cells induced with IPTG was observed on polyacrylamide gel electrophoresis (Fig. 4).
References
1. Fricke, J., Blei, F. & Hoffmeister, D. Enzymatic Synthesis of Psilocybin. Angew Chem Int Ed Engl 56, 12352-12355 (2017). 2. Artimo, P. et al. ExPASy: SIB bioinformatics resource portal. Nucleic Acids Res 40, W597-603 (2012).
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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