Part:BBa_K4249000
The biosynthetic gene for ERG production from N.crassa that catalyzing the first enzymatic step
The genes we used here were codon-optimized for S. cerevisiae according to reference(van der Hoek, Darbani et al. 2019). In nature, ergothioneine is produced through two different pathways: the bacterial pathway and the fungal pathway. In bacteria, the reaction sequence is catalyzed by five enzymes, encoded by egtA-E genes arranged in an operon. But in fungi, the biosynthetic pathway is different, only two genes are needed here, egt1 and egt2. Comparing two catalytic pathways, fungal pathway is simpler and more efficient. Since S.cerevisiae cannot synthesize ergothioneine directly, we applied CRISPR-Cas9 technology to integrate two heterologous genes into the yeast genome, and these two genes are NcEgt1 and CpEgt2 . Egt1 is a nonheme iron enzyme that could simultaneously catalyze two reactions, including the methylation of histidine to hecynine and the conversion of hecynine to hercynylcysteine sulfoxide. After that, another enzyme called Egt2, hercynylcysteine S-oxide lyase, will combine with hercynylcysteine sulfoxide to form EGT.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 751
Illegal EcoRI site found at 2078
Illegal XbaI site found at 1435 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 751
Illegal EcoRI site found at 2078 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 751
Illegal EcoRI site found at 2078
Illegal BglII site found at 613
Illegal BglII site found at 1099
Illegal BglII site found at 1432
Illegal BglII site found at 2101 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 751
Illegal EcoRI site found at 2078
Illegal XbaI site found at 1435 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 751
Illegal EcoRI site found at 2078
Illegal XbaI site found at 1435 - 1000COMPATIBLE WITH RFC[1000]
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