Biotransformation of hyoscyamine by a recombinant Saccharomyces cerevisiae strain

CARDILLO, ALEJANDRA BEATRIZ; SILVERSTEIN, SUSANA; GIULIETTI, ANA MARIA; MARCONI, PATRICIA

Rio de Janeiro

Congreso; ENPROMER 2nd Mercosur Congress on Chemical Engineering and 4th Congress on Process Systems Engineerig; 2005

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 6b-hydroxylase (H6H). We reported the isolation of h6h cDNA from B. candida group of tropane alkaloids including scopolamine. This biocompound is synthesised from hyoscyamine by action of Hyoscyamine 6b-hydroxylase (H6H). We reported the isolation of h6h cDNA from B. 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 6b-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. 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. (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. 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. 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. 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. (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. 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. 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. 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. (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. 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. 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. 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. (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. 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. 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. 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. (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. 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. 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. 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. (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. 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. 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. 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. (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. 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. 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. 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. (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. 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. 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. 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. (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. 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. 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. 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. (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. 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. 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. 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. (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. 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. 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. 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. (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. 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. 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. 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. (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. 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. 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. 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. (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. 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. 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. 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. (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. 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. 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. 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. (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. 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. 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. (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. 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. 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. 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. (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. 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. 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. 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. (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. 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. 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. 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. (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. 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. 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. 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. (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. 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. 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. 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. (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. 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. 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. 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. (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. 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. 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. 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. (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. 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. 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. (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. 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. 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. 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. (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. 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. 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. 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. (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. 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. 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. 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. (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. transformation. The recombinant strains were g