CARDILLO Alejandra Beatriz
congresos y reuniones científicas
BIOTRANSFORMATION AS A STRATEGY FOR THE PRODUCTION OF BIOACTIVE COMPOUNDS: TROPANE ALKALOIDS
CARDILLO, ALEJANDRA BEATRIZ; SARTUQUI, MARIELA; CARDILLO, SABRINA BEATRIZ; RODRIGUEZ TALOU, JULIAN; GIULIETTI, ANA MARIA
Congreso; REDBIO 2013; 2013
Tropane alkaloids, such as hyoscyamine, 6b-hydroxyhyoscyamine and scopolamine, were traditionally applied in medicine according to their anticholinergic activity. Hyoscyamine is converted by Hyoscyamine-6b-hydroxylase (H6H) into the most valuable alkaloids, 6b-hydroxyhyoscyamine and scopolamine. These bioactive compounds are obtained from producer plants due to the cost and complexity of their chemical synthesis. For this reason, its production by biotransformation is an attractive strategy for the pharmaceutical industry. In this work we explored the production of tropane alkaloids using the H6H as biocatalyst. The h6hgene was isolated from Brugmansia candida and cloned into Saccharomyces cerevisiae and Escherichia coli expression vectors in order to produce tagged and untagged enzymes. The enzyme produced was able to transform hyoscyamine into different products, showing a functional expression of the h6hcDNA. In yeast, the untagged H6H and the CBD-H6H were able to produce 6b-hydroxyhyoscyamine and scopolamine while H6H-V5-6His only produced 6b-hydroxyhyoscyamine. In addition, the Origami and Origami BL21 E. coli whole cells carrying the H6H were able to biotransform the alkaloid, producing 20% and 5% of 6b-hydroxyhyoscyamine, respectively. Further studies are needed in order to optimize and develop a biotransformation process of hyoscyamine using E. coli whole cells. However, these facts are encouraging for the development of a biocatalytic process using whole cells and plant enzymes free or immobilized.