Difference between revisions of "Part:BBa K3140001"
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The mechanism of psilocybin biosynthesis in ''Psilocybe'' sp. was recently elucidated by Fricke ''et al''.<ref name="Fricke">Fricke, J., Blei, F. & Hoffmeister, D. Enzymatic Synthesis of Psilocybin. ''Angew Chem Int Ed Engl'' '''56''', 12352-12355 (2017).</ref>, demonstrating that 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.
 
The mechanism of psilocybin biosynthesis in ''Psilocybe'' sp. was recently elucidated by Fricke ''et al''.<ref name="Fricke">Fricke, J., Blei, F. & Hoffmeister, D. Enzymatic Synthesis of Psilocybin. ''Angew Chem Int Ed Engl'' '''56''', 12352-12355 (2017).</ref>, demonstrating that 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.
  
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PsiH is a native enzyme obtained from ''Psilocybe cubensis'', which is involved in the metabolic biosynthesis of psilocybin from tryptophan. It accepts tryptamine as a substrate to yield 4-hydroxytryptamine ('''Fig. 1'''). 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<ref name="ExPASy">Artimo, P. et al. ExPASy: SIB bioinformatics resource portal. ''Nucleic Acids Res'' '''40''', W597-603 (2012).</ref>.
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PsiH is a native enzyme obtained from ''Psilocybe cubensis'', which is involved in the metabolic biosynthesis of psilocybin from tryptophan. It accepts tryptamine as a substrate to yield 4-hydroxytryptamine ('''Fig. 1'''). In a native state, PsiH is a 508 amino acid protein (57.5 kDa) with a theoretical pI of 5.94 calculated with the ExPASy ProtParam tool<ref name="ExPASy">Artimo, P. et al. ExPASy: SIB bioinformatics resource portal. ''Nucleic Acids Res'' '''40''', W597-603 (2012).</ref>.
  
 
[[Image:T--Sydney_Australia--PsiH_KEGG_rxn.gif|frame|none|Fig. 1: Hydroxylation of tryptamine to 4-hydroxytryptamine, mediated by PsiH. The reaction relies on a hydrogen donor (C00030) and O<sub>2</sub>, which results in the release of the oxidised donor (C00028) and H<sub>2</sub>O as a by-product. Source: [https://www.genome.jp/dbget-bin/www_bget?reaction+R11912 KEGG]  ]]
 
[[Image:T--Sydney_Australia--PsiH_KEGG_rxn.gif|frame|none|Fig. 1: Hydroxylation of tryptamine to 4-hydroxytryptamine, mediated by PsiH. The reaction relies on a hydrogen donor (C00030) and O<sub>2</sub>, which results in the release of the oxidised donor (C00028) and H<sub>2</sub>O as a by-product. Source: [https://www.genome.jp/dbget-bin/www_bget?reaction+R11912 KEGG]  ]]
  
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Heterologous expression of PsiH 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.
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Heterologous expression of PsiH has been achieved in a T7 induction system using   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.
  
  

Revision as of 03:18, 15 October 2019


PsiH - Cytochrome P450 monooxygenase from Psilocybe cubensis

PsiH is a cytochrome P450 monooxygenase that catalyses the conversion of tryptamine to 4-hydroxytryptamine.

Usage and Biology

The mechanism of psilocybin biosynthesis in Psilocybe sp. was recently elucidated by Fricke et al.[1], demonstrating that 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.

PsiH is a native enzyme obtained from Psilocybe cubensis, which is involved in the metabolic biosynthesis of psilocybin from tryptophan. It accepts tryptamine as a substrate to yield 4-hydroxytryptamine (Fig. 1). In a native state, PsiH is a 508 amino acid protein (57.5 kDa) with a theoretical pI of 5.94 calculated with the ExPASy ProtParam tool[2].

Fig. 1: Hydroxylation of tryptamine to 4-hydroxytryptamine, mediated by PsiH. The reaction relies on a hydrogen donor (C00030) and O2, which results in the release of the oxidised donor (C00028) and H2O as a by-product. Source: KEGG


Heterologous expression of PsiH has been achieved in a T7 induction system using 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.


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]


  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).