Coding

Part:BBa_K3140000

Designed by: Fahad Ali   Group: iGEM19_Sydney_Australia   (2019-10-12)
Revision as of 00:38, 15 October 2019 by Fahad3li (Talk | contribs)


PsiD - Tryptophan decarboxylase from Psilocybe cubensis

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

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. 3).

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: 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]


[edit]
Categories
Parameters
None