Part:BBa_K3140000

PsiD - Tryptophan decarboxylase from Psilocybe cubensis

PsiD is a tryptophan decarboxylase, which catalyses the conversion of L-tryptophan to tryptamine.

• NCBI: ASU62239.1
• UniProt: P0DPA6
• EC number: 4.1.1.105

Usage and Biology

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 and 4-hydroxytryptamine, respectively. 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. 4).

Heterologous expression of PsiD in pET-28c(+) yields a 475 amino acid polypeptide, with a computed molecular weight of 53550.71 Da.

Fig. 1: Decarboxylation of L-tryptophan to tryptamine, mediated by PsiD. CO₂ is released as a byproduct. Source: KEGG

Fig. 2: Decarboxylation of 4-hydroxy-L-tryptophan to 4-hydroxytryptamine, mediated by PsiD. CO₂ is released as a byproduct. Source: KEGG

Fig. 3: pET-28c(+):PsiD plasmid map, showing C-terminal histidine tag, and T7 promoter under the control of the lac operator. Translated peptide is shown as the thin lime green arrow.

Fig. 4: Polyacrylamide gel electrophoresis image of soluble and insoluble protein extract from uninduced and IPTG induced E. coli BL21(DE3)::pGro7 cells containing pET-28c(+):PsiD, run on an Mini-PROTEAN® TGX Stain-Free™ precast gel (Bio-Rad) at 200V for 40 minutes.

ADD:

- SDS-PAGE IMAGE - LC/MS - pUS250 and pET-28c(+)

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]