INVESTIGADORES
RODRIGUEZ TALOU Julian
congresos y reuniones científicas
Título:
Potential role of hyoscyamine 6 -h y d r o x y l a s e p l a n t e n z y m e i n pharmaceutical industry.
Autor/es:
CARDILLO AB; RODRÍGUEZ TALOU J; GIULIETTI AM.
Lugar:
Mar del Plata, Argentina
Reunión:
Congreso; XLIII Congreso de SAIB, Noviembre 2007, .; 2007
Institución organizadora:
SAIB
Resumen:
Plants produce compounds of interest in medicine. The South
American native plant, , produces tropane
alkaloids including hyoscyamine and scopolamine. The
anticholinergic properties of them made both alkaloids the most
important of the group. Scopolamine is widely used due to the fewer
side effects generated. Hyoscyamine 6 -hydroxylase (H6H, EC
1.14.11.11) is the enzyme responsible for the conversion of
hyoscyamine into scopolamine. In this work we report the
expression of H6H enzyme in as a
potential tool for the industrial production of scopolamine. The
cDNA was cloned into the pYES2 and the pYES2.1-TOPO TA
vectors. The constructions were introduced by chemical
transformation in CEN PK2. The enzyme was
expressed 4 hs after induction with galactose as it was observed in
Western blot analysis. The enzyme in vitro activity assay is being
carried out according to the bibliography described methods using
crude and IMAC semi-purified enzyme preparations. For the assay
analysis it was developed an HPLC method able to detect and
quantify low concentrations of alkaloids. It was possible to detect
less than 2 ppm of both alkaloids with a 2.6 resolution factor.