Cloning and Overexpression of Strictosidine β-D-Glucosidase GeneShort Sequence from Catharanthus Roseus in Escherichia coli

Purpose: Strictosidine-β-D-glucosidase (SGD) is considered as a key enzyme in the productionof bisindole alkaloids in Catharanthus roseus. The present study illustrated the production of ashort sequence of this enzyme in Escherichia coli without codon optimization.Methods: Strictosidine-β-D-glucosid...

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Published in:Advanced pharmaceutical bulletin 2019-10, Vol.9 (4), p.655-661
Main Authors: Arafa, Ahmed Saeed, Ragab, Amany Elsayed, Ibrahim, Abdel-Rahim Sayed, Abdel-Mageed, Wael Saad, Nasr, Mahmoud Emam
Format: Article
Language:English
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Summary:Purpose: Strictosidine-β-D-glucosidase (SGD) is considered as a key enzyme in the productionof bisindole alkaloids in Catharanthus roseus. The present study illustrated the production of ashort sequence of this enzyme in Escherichia coli without codon optimization.Methods: Strictosidine-β-D-glucosidase (sgd) gene short sequence (1434 bp), which lacksthe conserved sequence KGFFVWS and the localization peptide sequence at the C-terminal,was amplified from cDNA of C. roseus leaves, cloned and expressed in Escherichia coli. Theactivity of the produced protein in cell free lysate was tested using total alkaloid extract of C.roseus leaves.Results: HPLC and LC-MS analysis of the assay mixture revealed the disappearance of thestrictosidine peak.Conclusion: SGD short sequence can be produced in Escherichia coli in active form withoutcodon optimization.
ISSN:2228-5881
2251-7308
DOI:10.15171/apb.2019.076