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Expression of <it>Brugmansia candida </it>Hyoscyamine 6beta-Hydroxylase gene in <it>Saccharomyces cerevisiae </it>and its potential use as biocatalyst

By Giulietti Ana M, Talou Julián and Cardillo Alejandra B


<p>Abstract</p> <p>Background</p> <p>Tropane alkaloids, mainly hyoscyamine and scopolamine, are widely used in medicine due to their anticholinergic activity. Scopolamine has a higher demand being the more valuable alkaloid due to its fewer side effects and higher physiological activity. Anisodamine (6β-hydroxyhyoscyamine) is the intermediate in the conversion of hyoscyamine into scopolamine. Current studies report that this alkaloid is potentially applicable in medicine. The gene that codifies for Hyoscyamine 6-β hydroxylase, the enzyme responsible for hyoscyamine hydroxylation and epoxidation, leading to scopolamine was isolated from <it>Brugmansia candida</it>.</p> <p>Results</p> <p>The <it>h6h</it>cDNA was cloned into pYES2.1 and pYES2.1/V5-His-TOPO vectors to produce an untagged and a tagged protein, respectively. The H6H enzyme was produced in <it>Saccharomyces cerevisiae </it>in order to obtain a biological catalyst for potential industrial applications. Protein extracts of the induced yeast were analyzed by Western blot. The expression was detected 4 h after induction and no degradation was observed during the period assayed. The tagged and the untagged proteins were able to transform hyoscyamine, showing a functional expression of the <it>h6h</it>cDNA.</p> <p>Conclusion</p> <p>The strains obtained in this work are promising and potentially applicable in biocatalytic processes.</p

Topics: Microbiology, QR1-502, Science, Q, DOAJ:Microbiology, DOAJ:Biology, DOAJ:Biology and Life Sciences
Publisher: BioMed Central
Year: 2008
DOI identifier: 10.1186/1475-2859-7-17
OAI identifier:
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