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Analysis and sequencing of h6hmRNA, last enzyme in the tropane alkaloids pathway from anthers and hairy root cultures of Brugmansia candida (Solanaceae) Alejandra B.
Cardillo
Departamento Microbiología, Inmunología y Biotecnología Facultad de Farmacia y Bioquímica Universidad de Buenos Aires Junin 956 (1113) E-mail: alejandra_cardillo@yahoo.com Ana M. Giulietti Patricia L. Marconi* *Corresponding author Keywords: H6H, hairy roots, hyoscyamine, scopolamine, sequence analysis, transformation.
Brugmansia candida
(Solanaceae) is a native tree distributed across South-American
and produces the pharmacologically- important group of tropane alkaloids
including scopolamine. This biocompound is synthesised from hyoscyamine
by action of Hyoscyamine 6-β hydroxylase (H6H, EC 1.14.11.11)
at the end of the tropane alkaloid pathway. Here are reported the
tissue and organ-specific expression of h6hmRNA by RT-PCR analyses
and the isolation, cloning and sequencing of the cDNA obtained from
B. candida anthers and hairy root transformed cultures. Bioinformatic
analysis of the nucleotide sequence revealed an uninterrupted ORF
of 1038 bp and the predicted aminoacid sequence could be 344 aminoacid
long. A database search showed that this sequence has high homology
(97% identity) to Hyoscyamus |