CONTRATADOS
GIULIETTI Ana Maria
congresos y reuniones científicas
Título:
BIOTRANSFORMATION OF HYOSCYAMINE BY A RECOMBINANT
Autor/es:
CARDILLO A; GIULIETTI A
Lugar:
Río de Janeiro
Reunión:
Congreso; EMPROMER 2005; 2005
Institución organizadora:
Sociedad de Ingeniería Química de Brasil
Resumen:
Brugmansia candida is a South-American plant which produces the pharmacologically- important group of tropane alkaloids including scopolamine. This biocompound is synthesised from hyoscyamine by action of Hyoscyamine 6â-hydroxylase (H6H). We reported the isolation of h6h cDNA from B. candidais a South-American plant which produces the pharmacologically- important group of tropane alkaloids including scopolamine. This biocompound is synthesised from hyoscyamine by action of Hyoscyamine 6â-hydroxylase (H6H). We reported the isolation of h6h cDNA from B. candidaâ-hydroxylase (H6H). We reported the isolation of h6h cDNA from B. candida h6hmRNA by RT-PCR. Two isolated fragments (aprox. 1.1 kb) were cloned in Escherichia coli. The automated sequence and bioinformatic analyses revealed a high homology (97/98%) with the DNA sequence of h6hmRNA from other Solanaceae sps. The sequence of the insert has the ATG codon for further H6H expression. The aim of the present work is to obtain a recombinant S. cerevisiae strain expressing H6H isolated from B. candida for biotransformation purposes. Current experimental work is being performed in order to express B. candida H6H in S. cerevisiae. The H6H cDNA from B. candida was introduced under the control of GAL1 promoter in a pYES-TOPO vector (Invitrogen). The recombinant vectors were introduced into S. cerevisiae CEN.PK2 by chemical transformation. The recombinant strains were grown in SD without uracil for selection of transformants. Clones obtained are being assayed for their capacity for hyoscyamine biotransformation.Escherichia coli. The automated sequence and bioinformatic analyses revealed a high homology (97/98%) with the DNA sequence of h6hmRNA from other Solanaceae sps. The sequence of the insert has the ATG codon for further H6H expression. The aim of the present work is to obtain a recombinant S. cerevisiae strain expressing H6H isolated from B. candida for biotransformation purposes. Current experimental work is being performed in order to express B. candida H6H in S. cerevisiae. The H6H cDNA from B. candida was introduced under the control of GAL1 promoter in a pYES-TOPO vector (Invitrogen). The recombinant vectors were introduced into S. cerevisiae CEN.PK2 by chemical transformation. The recombinant strains were grown in SD without uracil for selection of transformants. Clones obtained are being assayed for their capacity for hyoscyamine biotransformation.S. cerevisiae strain expressing H6H isolated from B. candida for biotransformation purposes. Current experimental work is being performed in order to express B. candida H6H in S. cerevisiae. The H6H cDNA from B. candida was introduced under the control of GAL1 promoter in a pYES-TOPO vector (Invitrogen). The recombinant vectors were introduced into S. cerevisiae CEN.PK2 by chemical transformation. The recombinant strains were grown in SD without uracil for selection of transformants. Clones obtained are being assayed for their capacity for hyoscyamine biotransformation.for biotransformation purposes. Current experimental work is being performed in order to express B. candida H6H in S. cerevisiae. The H6H cDNA from B. candida was introduced under the control of GAL1 promoter in a pYES-TOPO vector (Invitrogen). The recombinant vectors were introduced into S. cerevisiae CEN.PK2 by chemical transformation. The recombinant strains were grown in SD without uracil for selection of transformants. Clones obtained are being assayed for their capacity for hyoscyamine biotransformation.B. candida H6H in S. cerevisiae. The H6H cDNA from B. candida was introduced under the control of GAL1 promoter in a pYES-TOPO vector (Invitrogen). The recombinant vectors were introduced into S. cerevisiae CEN.PK2 by chemical transformation. The recombinant strains were grown in SD without uracil for selection of transformants. Clones obtained are being assayed for their capacity for hyoscyamine biotransformation.B. candida was introduced under the control of GAL1 promoter in a pYES-TOPO vector (Invitrogen). The recombinant vectors were introduced into S. cerevisiae CEN.PK2 by chemical transformation. The recombinant strains were grown in SD without uracil for selection of transformants. Clones obtained are being assayed for their capacity for hyoscyamine biotransformation.S. cerevisiae CEN.PK2 by chemical transformation. The recombinant strains were grown in SD without uracil for selection of transformants. Clones obtained are being assayed for their capacity for hyoscyamine biotransformation.