Difference between revisions of "Part:BBa K3140004"
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===Usage and Biology===
 
===Usage and Biology===
  
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PsiH (codon optimised) is an enzyme derived from ''Psilocybe cubensis'', which is involved in the metabolic biosynthesis of psilocybin from tryptophan. The coding sequence has been codon optimised for expression in ''Escherichia coli''. It accepts tryptamine as a substrate to yield 4-hydroxytryptamine, respectively. Heterologous expression of PsiH has been achieved in ''Escherichia coli'' <add strain> in a <expression system>.
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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.
  
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[[Image:T--Sydney_Australia--PsiD_KEGG_rxn1.gif|thumb|900px|Fig. 1: Decarboxylation of L-tryptophan to tryptamine, mediated by PsiD. CO<sub>2</sub> is released as a byproduct. Source: [https://www.genome.jp/dbget-bin/www_bget?reaction+R00685 KEGG]  ]]
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PsiH (codon optimised) is an enzyme derived from ''Psilocybe cubensis'', which is involved in the metabolic biosynthesis of psilocybin from tryptophan. The coding sequence has been codon optimised for expression in ''Escherichia coli''. 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>.
  
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[[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 '''pCW (MORE)'''  transformed into ''Escherichia coli'' '''BL21(DE3)''', co-transformed with chaperone plasmid pGro7 ('''Fig. 3''').
  
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[[Image:T--Sydney_Australia--PsiD_KEGG_rxn2.gif|thumb|900px|Fig. 2: Decarboxylation of 4-hydroxy-L-tryptophan to 4-hydroxytryptamine, mediated by PsiD. CO<sub>2</sub> is released as a byproduct. Source: [https://www.genome.jp/dbget-bin/www_bget?reaction+R11932 KEGG]  ]]
 
  
  
 
<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K3140004 SequenceAndFeatures</partinfo>
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<partinfo>BBa_K3140001 SequenceAndFeatures</partinfo>
  
  
 
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===Functional Parameters===
 
===Functional Parameters===
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<partinfo>BBa_K3140004 parameters</partinfo>
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<partinfo>BBa_K3140001 parameters</partinfo>
 
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Revision as of 06:31, 15 October 2019


PsiH (codon optimised) - Cytochrome P450 monooxygenase from Psilocybe cubensis

PsiH (codon optimised) is a cytochrome P450 monooxygenase, which 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 (codon optimised) is an enzyme derived from Psilocybe cubensis, which is involved in the metabolic biosynthesis of psilocybin from tryptophan. The coding sequence has been codon optimised for expression in Escherichia coli. 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 pCW (MORE) transformed into Escherichia coli BL21(DE3), co-transformed with chaperone plasmid pGro7 (Fig. 3).


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).