INVESTIGADORES
RODRIGUEZ TALOU Julian
congresos y reuniones científicas
Título:
Hyoscyamine 6-â-hydroxylase, a recombinant biocatalyst for the industrial production of scopolamine
Autor/es:
CARDILLO AB; RODRÍGUEZ TALOU J; GIULIETTI AM
Lugar:
Dalian, China
Reunión:
Simposio; 13th International Biotechnology Symposium and Meeting; 2008
Institución organizadora:
International Biotechnology Symposium
Resumen:
The tropane alkaloids, hyoscyamine and scopolamine are widely
used as pharmaceuticals due to their anticholinergic activity.
Scopolamine is the more valuable, with a demand 10 times
higher than that of hyoscyamine (Palazón et al., 2003; Hashimoto
and Yamada, 1986). Brugmansia candida is a South American
native plant and a tropane alkaloid producer. The conversion of
hyoscyamine into scopolamine is carried out by hyoscyamine 6--
hydroxylase (H6H, EC 1.14.11.11). The development of a recombinant
cell harbouring the plant enzyme constitutes a useful strategy
for performing pharmaceutical processes. This work reports the
functional expression of H6H enzyme in Saccharomyces cerevisiae
native plant and a tropane alkaloid producer. The conversion of
hyoscyamine into scopolamine is carried out by hyoscyamine 6--
hydroxylase (H6H, EC 1.14.11.11). The development of a recombinant
cell harbouring the plant enzyme constitutes a useful strategy
for performing pharmaceutical processes. This work reports the
functional expression of H6H enzyme in Saccharomyces cerevisiae
and Yamada, 1986). Brugmansia candida is a South American
native plant and a tropane alkaloid producer. The conversion of
hyoscyamine into scopolamine is carried out by hyoscyamine 6--
hydroxylase (H6H, EC 1.14.11.11). The development of a recombinant
cell harbouring the plant enzyme constitutes a useful strategy
for performing pharmaceutical processes. This work reports the
functional expression of H6H enzyme in Saccharomyces cerevisiae
native plant and a tropane alkaloid producer. The conversion of
hyoscyamine into scopolamine is carried out by hyoscyamine 6--
hydroxylase (H6H, EC 1.14.11.11). The development of a recombinant
cell harbouring the plant enzyme constitutes a useful strategy
for performing pharmaceutical processes. This work reports the
functional expression of H6H enzyme in Saccharomyces cerevisiae
Palazón et al., 2003; Hashimoto
and Yamada, 1986). Brugmansia candida is a South American
native plant and a tropane alkaloid producer. The conversion of
hyoscyamine into scopolamine is carried out by hyoscyamine 6--
hydroxylase (H6H, EC 1.14.11.11). The development of a recombinant
cell harbouring the plant enzyme constitutes a useful strategy
for performing pharmaceutical processes. This work reports the
functional expression of H6H enzyme in Saccharomyces cerevisiae
native plant and a tropane alkaloid producer. The conversion of
hyoscyamine into scopolamine is carried out by hyoscyamine 6--
hydroxylase (H6H, EC 1.14.11.11). The development of a recombinant
cell harbouring the plant enzyme constitutes a useful strategy
for performing pharmaceutical processes. This work reports the
functional expression of H6H enzyme in Saccharomyces cerevisiae
). Brugmansia candida is a South American
native plant and a tropane alkaloid producer. The conversion of
hyoscyamine into scopolamine is carried out by hyoscyamine 6--
hydroxylase (H6H, EC 1.14.11.11). The development of a recombinant
cell harbouring the plant enzyme constitutes a useful strategy
for performing pharmaceutical processes. This work reports the
functional expression of H6H enzyme in Saccharomyces cerevisiaeSaccharomyces cerevisiae
as a potential tool for the industrial production of scopolamine.
The gene that codifies for the H6H enzyme was amplified from
B. candida immature anthers. The h6h cDNA obtained was cloned
into the pYES2 and the pYES2.1-TOPO TA vectors to produce an
untagged and a tagged enzyme, respectively. The constructions
were introduced in S. cerevisiae CEN PK2. The recombinant yeast
strains obtained were induced for the expression with galactose.
It was seen that the expression of the recombinant protein starts
4 h after induction. Crude protein extracts of the induced strains
were assayed for the enzyme activity (Hashimoto and Yamada,
1987; Liu et al., 2005). The activity assay was incubated at 30 ◦C
for 15 h. The analysis of the alkaloids was carried out by HPLC. The
mobile phase used was octanesulfonic acid 0.01M pH 3/methanol
(65:35), flow rate 1 ml/min. The results showed that the tagged
and untagged enzymeswere able to transform hyoscyamine, showing
a functional expression of the h6hcDNA. The untagged enzyme
presented a higher rate of conversion of hyoscyamine than the
tagged enzyme and was able to produce scopolamine and not only
6--hydroxyhyoscyamine in the incubation times assayed. It can
be concluded that the recombinant S. cerevisiae strains obtained
are promissory for technological applications in the production of
scopolamine by biotransformation.
are promissory for technological applications in the production of
scopolamine by biotransformation.
presented a higher rate of conversion of hyoscyamine than the
tagged enzyme and was able to produce scopolamine and not only
6--hydroxyhyoscyamine in the incubation times assayed. It can
be concluded that the recombinant S. cerevisiae strains obtained
are promissory for technological applications in the production of
scopolamine by biotransformation.
are promissory for technological applications in the production of
scopolamine by biotransformation.
for 15 h. The analysis of the alkaloids was carried out by HPLC. The
mobile phase used was octanesulfonic acid 0.01M pH 3/methanol
(65:35), flow rate 1 ml/min. The results showed that the tagged
and untagged enzymeswere able to transform hyoscyamine, showing
a functional expression of the h6hcDNA. The untagged enzyme
presented a higher rate of conversion of hyoscyamine than the
tagged enzyme and was able to produce scopolamine and not only
6--hydroxyhyoscyamine in the incubation times assayed. It can
be concluded that the recombinant S. cerevisiae strains obtained
are promissory for technological applications in the production of
scopolamine by biotransformation.
are promissory for technological applications in the production of
scopolamine by biotransformation.
presented a higher rate of conversion of hyoscyamine than the
tagged enzyme and was able to produce scopolamine and not only
6--hydroxyhyoscyamine in the incubation times assayed. It can
be concluded that the recombinant S. cerevisiae strains obtained
are promissory for technological applications in the production of
scopolamine by biotransformation.
are promissory for technological applications in the production of
scopolamine by biotransformation.
1987; Liu et al., 2005). The activity assay was incubated at 30 ◦C
for 15 h. The analysis of the alkaloids was carried out by HPLC. The
mobile phase used was octanesulfonic acid 0.01M pH 3/methanol
(65:35), flow rate 1 ml/min. The results showed that the tagged
and untagged enzymeswere able to transform hyoscyamine, showing
a functional expression of the h6hcDNA. The untagged enzyme
presented a higher rate of conversion of hyoscyamine than the
tagged enzyme and was able to produce scopolamine and not only
6--hydroxyhyoscyamine in the incubation times assayed. It can
be concluded that the recombinant S. cerevisiae strains obtained
are promissory for technological applications in the production of
scopolamine by biotransformation.
are promissory for technological applications in the production of
scopolamine by biotransformation.
presented a higher rate of conversion of hyoscyamine than the
tagged enzyme and was able to produce scopolamine and not only
6--hydroxyhyoscyamine in the incubation times assayed. It can
be concluded that the recombinant S. cerevisiae strains obtained
are promissory for technological applications in the production of
scopolamine by biotransformation.
are promissory for technological applications in the production of
scopolamine by biotransformation.
for 15 h. The analysis of the alkaloids was carried out by HPLC. The
mobile phase used was octanesulfonic acid 0.01M pH 3/methanol
(65:35), flow rate 1 ml/min. The results showed that the tagged
and untagged enzymeswere able to transform hyoscyamine, showing
a functional expression of the h6hcDNA. The untagged enzyme
presented a higher rate of conversion of hyoscyamine than the
tagged enzyme and was able to produce scopolamine and not only
6--hydroxyhyoscyamine in the incubation times assayed. It can
be concluded that the recombinant S. cerevisiae strains obtained
are promissory for technological applications in the production of
scopolamine by biotransformation.
are promissory for technological applications in the production of
scopolamine by biotransformation.
presented a higher rate of conversion of hyoscyamine than the
tagged enzyme and was able to produce scopolamine and not only
6--hydroxyhyoscyamine in the incubation times assayed. It can
be concluded that the recombinant S. cerevisiae strains obtained
are promissory for technological applications in the production of
scopolamine by biotransformation.
are promissory for technological applications in the production of
scopolamine by biotransformation.
strains obtained were induced for the expression with galactose.
It was seen that the expression of the recombinant protein starts
4 h after induction. Crude protein extracts of the induced strains
were assayed for the enzyme activity (Hashimoto and Yamada,
1987; Liu et al., 2005). The activity assay was incubated at 30 ◦C
for 15 h. The analysis of the alkaloids was carried out by HPLC. The
mobile phase used was octanesulfonic acid 0.01M pH 3/methanol
(65:35), flow rate 1 ml/min. The results showed that the tagged
and untagged enzymeswere able to transform hyoscyamine, showing
a functional expression of the h6hcDNA. The untagged enzyme
presented a higher rate of conversion of hyoscyamine than the
tagged enzyme and was able to produce scopolamine and not only
6--hydroxyhyoscyamine in the incubation times assayed. It can
be concluded that the recombinant S. cerevisiae strains obtained
are promissory for technological applications in the production of
scopolamine by biotransformation.
are promissory for technological applications in the production of
scopolamine by biotransformation.
presented a higher rate of conversion of hyoscyamine than the
tagged enzyme and was able to produce scopolamine and not only
6--hydroxyhyoscyamine in the incubation times assayed. It can
be concluded that the recombinant S. cerevisiae strains obtained
are promissory for technological applications in the production of
scopolamine by biotransformation.
are promissory for technological applications in the production of
scopolamine by biotransformation.
for 15 h. The analysis of the alkaloids was carried out by HPLC. The
mobile phase used was octanesulfonic acid 0.01M pH 3/methanol
(65:35), flow rate 1 ml/min. The results showed that the tagged
and untagged enzymeswere able to transform hyoscyamine, showing
a functional expression of the h6hcDNA. The untagged enzyme
presented a higher rate of conversion of hyoscyamine than the
tagged enzyme and was able to produce scopolamine and not only
6--hydroxyhyoscyamine in the incubation times assayed. It can
be concluded that the recombinant S. cerevisiae strains obtained
are promissory for technological applications in the production of
scopolamine by biotransformation.
are promissory for technological applications in the production of
scopolamine by biotransformation.
presented a higher rate of conversion of hyoscyamine than the
tagged enzyme and was able to produce scopolamine and not only
6--hydroxyhyoscyamine in the incubation times assayed. It can
be concluded that the recombinant S. cerevisiae strains obtained
are promissory for technological applications in the production of
scopolamine by biotransformation.
are promissory for technological applications in the production of
scopolamine by biotransformation.
1987; Liu et al., 2005). The activity assay was incubated at 30 ◦C
for 15 h. The analysis of the alkaloids was carried out by HPLC. The
mobile phase used was octanesulfonic acid 0.01M pH 3/methanol
(65:35), flow rate 1 ml/min. The results showed that the tagged
and untagged enzymeswere able to transform hyoscyamine, showing
a functional expression of the h6hcDNA. The untagged enzyme
presented a higher rate of conversion of hyoscyamine than the
tagged enzyme and was able to produce scopolamine and not only
6--hydroxyhyoscyamine in the incubation times assayed. It can
be concluded that the recombinant S. cerevisiae strains obtained
are promissory for technological applications in the production of
scopolamine by biotransformation.
are promissory for technological applications in the production of
scopolamine by biotransformation.
presented a higher rate of conversion of hyoscyamine than the
tagged enzyme and was able to produce scopolamine and not only
6--hydroxyhyoscyamine in the incubation times assayed. It can
be concluded that the recombinant S. cerevisiae strains obtained
are promissory for technological applications in the production of
scopolamine by biotransformation.
are promissory for technological applications in the production of
scopolamine by biotransformation.
for 15 h. The analysis of the alkaloids was carried out by HPLC. The
mobile phase used was octanesulfonic acid 0.01M pH 3/methanol
(65:35), flow rate 1 ml/min. The results showed that the tagged
and untagged enzymeswere able to transform hyoscyamine, showing
a functional expression of the h6hcDNA. The untagged enzyme
presented a higher rate of conversion of hyoscyamine than the
tagged enzyme and was able to produce scopolamine and not only
6--hydroxyhyoscyamine in the incubation times assayed. It can
be concluded that the recombinant S. cerevisiae strains obtained
are promissory for technological applications in the production of
scopolamine by biotransformation.
are promissory for technological applications in the production of
scopolamine by biotransformation.
presented a higher rate of conversion of hyoscyamine than the
tagged enzyme and was able to produce scopolamine and not only
6--hydroxyhyoscyamine in the incubation times assayed. It can
be concluded that the recombinant S. cerevisiae strains obtained
are promissory for technological applications in the production of
scopolamine by biotransformation.
are promissory for technological applications in the production of
scopolamine by biotransformation.
into the pYES2 and the pYES2.1-TOPO TA vectors to produce an
untagged and a tagged enzyme, respectively. The constructions
were introduced in S. cerevisiae CEN PK2. The recombinant yeast
strains obtained were induced for the expression with galactose.
It was seen that the expression of the recombinant protein starts
4 h after induction. Crude protein extracts of the induced strains
were assayed for the enzyme activity (Hashimoto and Yamada,
1987; Liu et al., 2005). The activity assay was incubated at 30 ◦C
for 15 h. The analysis of the alkaloids was carried out by HPLC. The
mobile phase used was octanesulfonic acid 0.01M pH 3/methanol
(65:35), flow rate 1 ml/min. The results showed that the tagged
and untagged enzymeswere able to transform hyoscyamine, showing
a functional expression of the h6hcDNA. The untagged enzyme
presented a higher rate of conversion of hyoscyamine than the
tagged enzyme and was able to produce scopolamine and not only
6--hydroxyhyoscyamine in the incubation times assayed. It can
be concluded that the recombinant S. cerevisiae strains obtained
are promissory for technological applications in the production of
scopolamine by biotransformation.
are promissory for technological applications in the production of
scopolamine by biotransformation.
presented a higher rate of conversion of hyoscyamine than the
tagged enzyme and was able to produce scopolamine and not only
6--hydroxyhyoscyamine in the incubation times assayed. It can
be concluded that the recombinant S. cerevisiae strains obtained
are promissory for technological applications in the production of
scopolamine by biotransformation.
are promissory for technological applications in the production of
scopolamine by biotransformation.
for 15 h. The analysis of the alkaloids was carried out by HPLC. The
mobile phase used was octanesulfonic acid 0.01M pH 3/methanol
(65:35), flow rate 1 ml/min. The results showed that the tagged
and untagged enzymeswere able to transform hyoscyamine, showing
a functional expression of the h6hcDNA. The untagged enzyme
presented a higher rate of conversion of hyoscyamine than the
tagged enzyme and was able to produce scopolamine and not only
6--hydroxyhyoscyamine in the incubation times assayed. It can
be concluded that the recombinant S. cerevisiae strains obtained
are promissory for technological applications in the production of
scopolamine by biotransformation.
are promissory for technological applications in the production of
scopolamine by biotransformation.
presented a higher rate of conversion of hyoscyamine than the
tagged enzyme and was able to produce scopolamine and not only
6--hydroxyhyoscyamine in the incubation times assayed. It can
be concluded that the recombinant S. cerevisiae strains obtained
are promissory for technological applications in the production of
scopolamine by biotransformation.
are promissory for technological applications in the production of
scopolamine by biotransformation.
1987; Liu et al., 2005). The activity assay was incubated at 30 ◦C
for 15 h. The analysis of the alkaloids was carried out by HPLC. The
mobile phase used was octanesulfonic acid 0.01M pH 3/methanol
(65:35), flow rate 1 ml/min. The results showed that the tagged
and untagged enzymeswere able to transform hyoscyamine, showing
a functional expression of the h6hcDNA. The untagged enzyme
presented a higher rate of conversion of hyoscyamine than the
tagged enzyme and was able to produce scopolamine and not only
6--hydroxyhyoscyamine in the incubation times assayed. It can
be concluded that the recombinant S. cerevisiae strains obtained
are promissory for technological applications in the production of
scopolamine by biotransformation.
are promissory for technological applications in the production of
scopolamine by biotransformation.
presented a higher rate of conversion of hyoscyamine than the
tagged enzyme and was able to produce scopolamine and not only
6--hydroxyhyoscyamine in the incubation times assayed. It can
be concluded that the recombinant S. cerevisiae strains obtained
are promissory for technological applications in the production of
scopolamine by biotransformation.
are promissory for technological applications in the production of
scopolamine by biotransformation.
for 15 h. The analysis of the alkaloids was carried out by HPLC. The
mobile phase used was octanesulfonic acid 0.01M pH 3/methanol
(65:35), flow rate 1 ml/min. The results showed that the tagged
and untagged enzymeswere able to transform hyoscyamine, showing
a functional expression of the h6hcDNA. The untagged enzyme
presented a higher rate of conversion of hyoscyamine than the
tagged enzyme and was able to produce scopolamine and not only
6--hydroxyhyoscyamine in the incubation times assayed. It can
be concluded that the recombinant S. cerevisiae strains obtained
are promissory for technological applications in the production of
scopolamine by biotransformation.
are promissory for technological applications in the production of
scopolamine by biotransformation.
presented a higher rate of conversion of hyoscyamine than the
tagged enzyme and was able to produce scopolamine and not only
6--hydroxyhyoscyamine in the incubation times assayed. It can
be concluded that the recombinant S. cerevisiae strains obtained
are promissory for technological applications in the production of
scopolamine by biotransformation.
are promissory for technological applications in the production of
scopolamine by biotransformation.
strains obtained were induced for the expression with galactose.
It was seen that the expression of the recombinant protein starts
4 h after induction. Crude protein extracts of the induced strains
were assayed for the enzyme activity (Hashimoto and Yamada,
1987; Liu et al., 2005). The activity assay was incubated at 30 ◦C
for 15 h. The analysis of the alkaloids was carried out by HPLC. The
mobile phase used was octanesulfonic acid 0.01M pH 3/methanol
(65:35), flow rate 1 ml/min. The results showed that the tagged
and untagged enzymeswere able to transform hyoscyamine, showing
a functional expression of the h6hcDNA. The untagged enzyme
presented a higher rate of conversion of hyoscyamine than the
tagged enzyme and was able to produce scopolamine and not only
6--hydroxyhyoscyamine in the incubation times assayed. It can
be concluded that the recombinant S. cerevisiae strains obtained
are promissory for technological applications in the production of
scopolamine by biotransformation.
are promissory for technological applications in the production of
scopolamine by biotransformation.
presented a higher rate of conversion of hyoscyamine than the
tagged enzyme and was able to produce scopolamine and not only
6--hydroxyhyoscyamine in the incubation times assayed. It can
be concluded that the recombinant S. cerevisiae strains obtained
are promissory for technological applications in the production of
scopolamine by biotransformation.
are promissory for technological applications in the production of
scopolamine by biotransformation.
for 15 h. The analysis of the alkaloids was carried out by HPLC. The
mobile phase used was octanesulfonic acid 0.01M pH 3/methanol
(65:35), flow rate 1 ml/min. The results showed that the tagged
and untagged enzymeswere able to transform hyoscyamine, showing
a functional expression of the h6hcDNA. The untagged enzyme
presented a higher rate of conversion of hyoscyamine than the
tagged enzyme and was able to produce scopolamine and not only
6--hydroxyhyoscyamine in the incubation times assayed. It can
be concluded that the recombinant S. cerevisiae strains obtained
are promissory for technological applications in the production of
scopolamine by biotransformation.
are promissory for technological applications in the production of
scopolamine by biotransformation.
presented a higher rate of conversion of hyoscyamine than the
tagged enzyme and was able to produce scopolamine and not only
6--hydroxyhyoscyamine in the incubation times assayed. It can
be concluded that the recombinant S. cerevisiae strains obtained
are promissory for technological applications in the production of
scopolamine by biotransformation.
are promissory for technological applications in the production of
scopolamine by biotransformation.
1987; Liu et al., 2005). The activity assay was incubated at 30 ◦C
for 15 h. The analysis of the alkaloids was carried out by HPLC. The
mobile phase used was octanesulfonic acid 0.01M pH 3/methanol
(65:35), flow rate 1 ml/min. The results showed that the tagged
and untagged enzymeswere able to transform hyoscyamine, showing
a functional expression of the h6hcDNA. The untagged enzyme
presented a higher rate of conversion of hyoscyamine than the
tagged enzyme and was able to produce scopolamine and not only
6--hydroxyhyoscyamine in the incubation times assayed. It can
be concluded that the recombinant S. cerevisiae strains obtained
are promissory for technological applications in the production of
scopolamine by biotransformation.
are promissory for technological applications in the production of
scopolamine by biotransformation.
presented a higher rate of conversion of hyoscyamine than the
tagged enzyme and was able to produce scopolamine and not only
6--hydroxyhyoscyamine in the incubation times assayed. It can
be concluded that the recombinant S. cerevisiae strains obtained
are promissory for technological applications in the production of
scopolamine by biotransformation.
are promissory for technological applications in the production of
scopolamine by biotransformation.
for 15 h. The analysis of the alkaloids was carried out by HPLC. The
mobile phase used was octanesulfonic acid 0.01M pH 3/methanol
(65:35), flow rate 1 ml/min. The results showed that the tagged
and untagged enzymeswere able to transform hyoscyamine, showing
a functional expression of the h6hcDNA. The untagged enzyme
presented a higher rate of conversion of hyoscyamine than the
tagged enzyme and was able to produce scopolamine and not only
6--hydroxyhyoscyamine in the incubation times assayed. It can
be concluded that the recombinant S. cerevisiae strains obtained
are promissory for technological applications in the production of
scopolamine by biotransformation.
are promissory for technological applications in the production of
scopolamine by biotransformation.
presented a higher rate of conversion of hyoscyamine than the
tagged enzyme and was able to produce scopolamine and not only
6--hydroxyhyoscyamine in the incubation times assayed. It can
be concluded that the recombinant S. cerevisiae strains obtained
are promissory for technological applications in the production of
scopolamine by biotransformation.
are promissory for technological applications in the production of
scopolamine by biotransformation.
immature anthers. The h6h cDNA obtained was cloned
into the pYES2 and the pYES2.1-TOPO TA vectors to produce an
untagged and a tagged enzyme, respectively. The constructions
were introduced in S. cerevisiae CEN PK2. The recombinant yeast
strains obtained were induced for the expression with galactose.
It was seen that the expression of the recombinant protein starts
4 h after induction. Crude protein extracts of the induced strains
were assayed for the enzyme activity (Hashimoto and Yamada,
1987; Liu et al., 2005). The activity assay was incubated at 30 ◦C
for 15 h. The analysis of the alkaloids was carried out by HPLC. The
mobile phase used was octanesulfonic acid 0.01M pH 3/methanol
(65:35), flow rate 1 ml/min. The results showed that the tagged
and untagged enzymeswere able to transform hyoscyamine, showing
a functional expression of the h6hcDNA. The untagged enzyme
presented a higher rate of conversion of hyoscyamine than the
tagged enzyme and was able to produce scopolamine and not only
6--hydroxyhyoscyamine in the incubation times assayed. It can
be concluded that the recombinant S. cerevisiae strains obtained
are promissory for technological applications in the production of
scopolamine by biotransformation.
are promissory for technological applications in the production of
scopolamine by biotransformation.
presented a higher rate of conversion of hyoscyamine than the
tagged enzyme and was able to produce scopolamine and not only
6--hydroxyhyoscyamine in the incubation times assayed. It can
be concluded that the recombinant S. cerevisiae strains obtained
are promissory for technological applications in the production of
scopolamine by biotransformation.
are promissory for technological applications in the production of
scopolamine by biotransformation.
for 15 h. The analysis of the alkaloids was carried out by HPLC. The
mobile phase used was octanesulfonic acid 0.01M pH 3/methanol
(65:35), flow rate 1 ml/min. The results showed that the tagged
and untagged enzymeswere able to transform hyoscyamine, showing
a functional expression of the h6hcDNA. The untagged enzyme
presented a higher rate of conversion of hyoscyamine than the
tagged enzyme and was able to produce scopolamine and not only
6--hydroxyhyoscyamine in the incubation times assayed. It can
be concluded that the recombinant S. cerevisiae strains obtained
are promissory for technological applications in the production of
scopolamine by biotransformation.
are promissory for technological applications in the production of
scopolamine by biotransformation.
presented a higher rate of conversion of hyoscyamine than the
tagged enzyme and was able to produce scopolamine and not only
6--hydroxyhyoscyamine in the incubation times assayed. It can
be concluded that the recombinant S. cerevisiae strains obtained
are promissory for technological applications in the production of
scopolamine by biotransformation.
are promissory for technological applications in the production of
scopolamine by biotransformation.
1987; Liu et al., 2005). The activity assay was incubated at 30 ◦C
for 15 h. The analysis of the alkaloids was carried out by HPLC. The
mobile phase used was octanesulfonic acid 0.01M pH 3/methanol
(65:35), flow rate 1 ml/min. The results showed that the tagged
and untagged enzymeswere able to transform hyoscyamine, showing
a functional expression of the h6hcDNA. The untagged enzyme
presented a higher rate of conversion of hyoscyamine than the
tagged enzyme and was able to produce scopolamine and not only
6--hydroxyhyoscyamine in the incubation times assayed. It can
be concluded that the recombinant S. cerevisiae strains obtained
are promissory for technological applications in the production of
scopolamine by biotransformation.
are promissory for technological applications in the production of
scopolamine by biotransformation.
presented a higher rate of conversion of hyoscyamine than the
tagged enzyme and was able to produce scopolamine and not only
6--hydroxyhyoscyamine in the incubation times assayed. It can
be concluded that the recombinant S. cerevisiae strains obtained
are promissory for technological applications in the production of
scopolamine by biotransformation.
are promissory for technological applications in the production of
scopolamine by biotransformation.
for 15 h. The analysis of the alkaloids was carried out by HPLC. The
mobile phase used was octanesulfonic acid 0.01M pH 3/methanol
(65:35), flow rate 1 ml/min. The results showed that the tagged
and untagged enzymeswere able to transform hyoscyamine, showing
a functional expression of the h6hcDNA. The untagged enzyme
presented a higher rate of conversion of hyoscyamine than the
tagged enzyme and was able to produce scopolamine and not only
6--hydroxyhyoscyamine in the incubation times assayed. It can
be concluded that the recombinant S. cerevisiae strains obtained
are promissory for technological applications in the production of
scopolamine by biotransformation.
are promissory for technological applications in the production of
scopolamine by biotransformation.
presented a higher rate of conversion of hyoscyamine than the
tagged enzyme and was able to produce scopolamine and not only
6--hydroxyhyoscyamine in the incubation times assayed. It can
be concluded that the recombinant S. cerevisiae strains obtained
are promissory for technological applications in the production of
scopolamine by biotransformation.
are promissory for technological applications in the production of
scopolamine by biotransformation.
strains obtained were induced for the expression with galactose.
It was seen that the expression of the recombinant protein starts
4 h after induction. Crude protein extracts of the induced strains
were assayed for the enzyme activity (Hashimoto and Yamada,
1987; Liu et al., 2005). The activity assay was incubated at 30 ◦C
for 15 h. The analysis of the alkaloids was carried out by HPLC. The
mobile phase used was octanesulfonic acid 0.01M pH 3/methanol
(65:35), flow rate 1 ml/min. The results showed that the tagged
and untagged enzymeswere able to transform hyoscyamine, showing
a functional expression of the h6hcDNA. The untagged enzyme
presented a higher rate of conversion of hyoscyamine than the
tagged enzyme and was able to produce scopolamine and not only
6--hydroxyhyoscyamine in the incubation times assayed. It can
be concluded that the recombinant S. cerevisiae strains obtained
are promissory for technological applications in the production of
scopolamine by biotransformation.
are promissory for technological applications in the production of
scopolamine by biotransformation.
presented a higher rate of conversion of hyoscyamine than the
tagged enzyme and was able to produce scopolamine and not only
6--hydroxyhyoscyamine in the incubation times assayed. It can
be concluded that the recombinant S. cerevisiae strains obtained
are promissory for technological applications in the production of
scopolamine by biotransformation.
are promissory for technological applications in the production of
scopolamine by biotransformation.
for 15 h. The analysis of the alkaloids was carried out by HPLC. The
mobile phase used was octanesulfonic acid 0.01M pH 3/methanol
(65:35), flow rate 1 ml/min. The results showed that the tagged
and untagged enzymeswere able to transform hyoscyamine, showing
a functional expression of the h6hcDNA. The untagged enzyme
presented a higher rate of conversion of hyoscyamine than the
tagged enzyme and was able to produce scopolamine and not only
6--hydroxyhyoscyamine in the incubation times assayed. It can
be concluded that the recombinant S. cerevisiae strains obtained
are promissory for technological applications in the production of
scopolamine by biotransformation.
are promissory for technological applications in the production of
scopolamine by biotransformation.
presented a higher rate of conversion of hyoscyamine than the
tagged enzyme and was able to produce scopolamine and not only
6--hydroxyhyoscyamine in the incubation times assayed. It can
be concluded that the recombinant S. cerevisiae strains obtained
are promissory for technological applications in the production of
scopolamine by biotransformation.
are promissory for technological applications in the production of
scopolamine by biotransformation.
1987; Liu et al., 2005). The activity assay was incubated at 30 ◦C
for 15 h. The analysis of the alkaloids was carried out by HPLC. The
mobile phase used was octanesulfonic acid 0.01M pH 3/methanol
(65:35), flow rate 1 ml/min. The results showed that the tagged
and untagged enzymeswere able to transform hyoscyamine, showing
a functional expression of the h6hcDNA. The untagged enzyme
presented a higher rate of conversion of hyoscyamine than the
tagged enzyme and was able to produce scopolamine and not only
6--hydroxyhyoscyamine in the incubation times assayed. It can
be concluded that the recombinant S. cerevisiae strains obtained
are promissory for technological applications in the production of
scopolamine by biotransformation.
are promissory for technological applications in the production of
scopolamine by biotransformation.
presented a higher rate of conversion of hyoscyamine than the
tagged enzyme and was able to produce scopolamine and not only
6--hydroxyhyoscyamine in the incubation times assayed. It can
be concluded that the recombinant S. cerevisiae strains obtained
are promissory for technological applications in the production of
scopolamine by biotransformation.
are promissory for technological applications in the production of
scopolamine by biotransformation.
for 15 h. The analysis of the alkaloids was carried out by HPLC. The
mobile phase used was octanesulfonic acid 0.01M pH 3/methanol
(65:35), flow rate 1 ml/min. The results showed that the tagged
and untagged enzymeswere able to transform hyoscyamine, showing
a functional expression of the h6hcDNA. The untagged enzyme
presented a higher rate of conversion of hyoscyamine than the
tagged enzyme and was able to produce scopolamine and not only
6--hydroxyhyoscyamine in the incubation times assayed. It can
be concluded that the recombinant S. cerevisiae strains obtained
are promissory for technological applications in the production of
scopolamine by biotransformation.
are promissory for technological applications in the production of
scopolamine by biotransformation.
presented a higher rate of conversion of hyoscyamine than the
tagged enzyme and was able to produce scopolamine and not only
6--hydroxyhyoscyamine in the incubation times assayed. It can
be concluded that the recombinant S. cerevisiae strains obtained
are promissory for technological applications in the production of
scopolamine by biotransformation.
are promissory for technological applications in the production of
scopolamine by biotransformation.
S. cerevisiae CEN PK2. The recombinant yeast
strains obtained were induced for the expression with galactose.
It was seen that the expression of the recombinant protein starts
4 h after induction. Crude protein extracts of the induced strains
were assayed for the enzyme activity (Hashimoto and Yamada,
1987; Liu et al., 2005). The activity assay was incubated at 30 ◦C
for 15 h. The analysis of the alkaloids was carried out by HPLC. The
mobile phase used was octanesulfonic acid 0.01M pH 3/methanol
(65:35), flow rate 1 ml/min. The results showed that the tagged
and untagged enzymeswere able to transform hyoscyamine, showing
a functional expression of the h6hcDNA. The untagged enzyme
presented a higher rate of conversion of hyoscyamine than the
tagged enzyme and was able to produce scopolamine and not only
6--hydroxyhyoscyamine in the incubation times assayed. It can
be concluded that the recombinant S. cerevisiae strains obtained
are promissory for technological applications in the production of
scopolamine by biotransformation.
are promissory for technological applications in the production of
scopolamine by biotransformation.
presented a higher rate of conversion of hyoscyamine than the
tagged enzyme and was able to produce scopolamine and not only
6--hydroxyhyoscyamine in the incubation times assayed. It can
be concluded that the recombinant S. cerevisiae strains obtained
are promissory for technological applications in the production of
scopolamine by biotransformation.
are promissory for technological applications in the production of
scopolamine by biotransformation.
for 15 h. The analysis of the alkaloids was carried out by HPLC. The
mobile phase used was octanesulfonic acid 0.01M pH 3/methanol
(65:35), flow rate 1 ml/min. The results showed that the tagged
and untagged enzymeswere able to transform hyoscyamine, showing
a functional expression of the h6hcDNA. The untagged enzyme
presented a higher rate of conversion of hyoscyamine than the
tagged enzyme and was able to produce scopolamine and not only
6--hydroxyhyoscyamine in the incubation times assayed. It can
be concluded that the recombinant S. cerevisiae strains obtained
are promissory for technological applications in the production of
scopolamine by biotransformation.
are promissory for technological applications in the production of
scopolamine by biotransformation.
presented a higher rate of conversion of hyoscyamine than the
tagged enzyme and was able to produce scopolamine and not only
6--hydroxyhyoscyamine in the incubation times assayed. It can
be concluded that the recombinant S. cerevisiae strains obtained
are promissory for technological applications in the production of
scopolamine by biotransformation.
are promissory for technological applications in the production of
scopolamine by biotransformation.
1987; Liu et al., 2005). The activity assay was incubated at 30 ◦C
for 15 h. The analysis of the alkaloids was carried out by HPLC. The
mobile phase used was octanesulfonic acid 0.01M pH 3/methanol
(65:35), flow rate 1 ml/min. The results showed that the tagged
and untagged enzymeswere able to transform hyoscyamine, showing
a functional expression of the h6hcDNA. The untagged enzyme
presented a higher rate of conversion of hyoscyamine than the
tagged enzyme and was able to produce scopolamine and not only
6--hydroxyhyoscyamine in the incubation times assayed. It can
be concluded that the recombinant S. cerevisiae strains obtained
are promissory for technological applications in the production of
scopolamine by biotransformation.
are promissory for technological applications in the production of
scopolamine by biotransformation.
presented a higher rate of conversion of hyoscyamine than the
tagged enzyme and was able to produce scopolamine and not only
6--hydroxyhyoscyamine in the incubation times assayed. It can
be concluded that the recombinant S. cerevisiae strains obtained
are promissory for technological applications in the production of
scopolamine by biotransformation.