Expression of Brugmansia candida Hyoscyamine 6beta-Hydroxylase gene in Saccharomyces cerevisiae and its potential use as biocatalyst

Cardillo, Alejandra B.; Talou, Julián Rodríguez; Giulietti, Ana María

doi:10.1186/1475-2859-7-17

Cited by 31 publications

(26 citation statements)

References 27 publications

(28 reference statements)

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“…Consistent with our findings, a similar phenomenon was previously reported in H. niger (Hashimoto et al 1993). Recently, a H6H gene from Brugmansia candida was functionally expressed in yeast indicating a good potential for use as a biocatalyst (Cardillo et al 2008). Because the complex structures of alkaloids make chemical synthesis complicated and expensive, our genetically engineered bacterial strains which can express bioactive product provide an alternative promising bioreactor to produce scopolamine as a new biocatalyst.…”

Section: Bioconversion Of Hyoscyamine To Scopolamine In E Colisupporting

confidence: 90%

Functional identification of hyoscyamine 6β-hydroxylase from Anisodus acutangulus and overproduction of scopolamine in genetically-engineered Escherichia coli

et al. 2011

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Hyoscyamine 6β-hydroxylase (H6H; EC 1.14.11.11) converts hyoscyamine to scopolamine in the last step of scopolamine biosynthetic pathway. The gene encoding H6H in Anisodus acutangulus was cloned and expressed in Escherichia coli and the recombinant proteins fused with His-tag or GST-tag at its N-terminal were purified and then confirmed by Western bolt analysis. The biofunctional assay revealed that the His-AaH6H and GST-AaH6H converted hyoscyamine (40 mg/l) to scopolamine at 32 and 31 mg/l, respectively. This is the first report on AaH6H expression, purification and functional characterization facilitates further genetic improvement of scopolamine yield in A. acutangulus.

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Section: Bioconversion Of Hyoscyamine To Scopolamine In E Colisupporting

confidence: 90%

Functional identification of hyoscyamine 6β-hydroxylase from Anisodus acutangulus and overproduction of scopolamine in genetically-engineered Escherichia coli

et al. 2011

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“…When the two varieties are compared, it can be seen that the H6H hydroxylase and epoxidase activities of the Argentinean clones are significantly higher than that of the Colombian clones (p = 0.0094 and 0.0265, respectively). Hyoscyamine (1) biotransformation processes carried out with a recombinant Saccharomyces cerevisiae strain carrying the h6h cDNA obtained from Argentinean B. candida plants showed similar results, probably related to the nature of the isolated gene [22,23]. Cardillo et al [23] reported that the recombinant en- zyme converted mainly hyoscyamine into anisodamine.…”

Section: Dwmentioning

confidence: 83%

“…Hyoscyamine (1) biotransformation processes carried out with a recombinant Saccharomyces cerevisiae strain carrying the h6h cDNA obtained from Argentinean B. candida plants showed similar results, probably related to the nature of the isolated gene [22,23]. Cardillo et al [23] reported that the recombinant en- zyme converted mainly hyoscyamine into anisodamine. The specific hydroxylase activity measured in yeast extracts was 10 times higher than the epoxidase activity.…”

Section: Dwmentioning

confidence: 83%

Anisodamine Production from Natural Sources: Seedlings and Hairy Root Cultures of Argentinean and ColombianBrugmansia candidaPlants

Cardillo¹,

Alvarez²,

Lopez³

et al. 2009

Planta Med

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The tropane alkaloid anisodamine ( 2) is obtained by 6 beta-hydroxylation of hyoscyamine ( 1). The application of this alkaloid in medicine is gaining attention due to the wide range of therapeutic applications described in addition to its anticholinergic activity. In this work, the production of anisodamine ( 2) by IN VITRO cultures of BRUGMANSIA CANDIDA (Argentinean and Colombian samples) was studied. This alkaloid was estimated in different organs of IN VITRO-germinated seedlings as well as in hairy roots obtained from seedlings from both sources. Colombian roots exhibited the highest content of tropane alkaloids, with anisodamine ( 2) being the main alkaloid measured. In the leaves, the main alkaloid was scopolamine ( 3) and no significant differences were observed between Argentinean and Colombian leaves. The tropane alkaloid content in Argentinean hairy roots was significantly higher than in Colombian ones. Also, in the Argentinean samples the main alkaloid detected was anisodamine ( 2). Argentinean and Colombian B. CANDIDA seedlings and hairy roots appear to be a promising system for the production of anisodamine ( 2).

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“…Acetyltropic acid (3-acetoxy-2-phenylpropanoic acid) is an important intermediate for synthesis of tropane alkaloids, such as DL-hyoscyamine [1], L-hyoscyamine [2,3], anisodamine [4][5][6], scopolamine [4,7], 6,7-dehydrohyoscyamine [7,8], and anisodine [9]. Stereochemical structure of acetyltropic acid is shown in Figure 1.…”

Section: Introductionmentioning

confidence: 99%

Enantioseparation of acetyltropic acid by countercurrent chromatography with sulfobutyl ether‐β‐cyclodextrin as chiral selector

Qiu

Sun

Wang

et al. 2019

J of Separation Science

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Acetyltropic acid is an important synthetic intermediate for preparation of tropane alkaloid derivatives, which can be used as anticholinergic drugs, deliriants, and stimulants. In the present work, acetyltropic acid was successfully enantioseparated by countercurrent chromatography using sulfobutyl ether-β-cyclodextrin as chiral selector. A biphasic solvent system composed of n-butyl acetate/n-hexane/0.1 mol/L citrate buffer at pH = 2.2 containing 0.1 mol/L of sulfobutyl ether-β-cyclodextrin (7:3:10, v/v) was selected, which produced a suitable distribution ratio D S = 1.14, D R = 2.31 and a high enantioseparation factor α = 2.03. Baseline separation was achieved for preparative enantioseparation of 50 mg of racemic acetyltropic acid. A method for chiral analysis of acetyltropic acid by conventional reverse phase liquid chromatography with hydroxylpropyl-β-cyclodextrin as mobile phase additive was established, and formation constants of inclusion complex were determined. It was found that different substituted β-cyclodextrin should be selected for enantioseparation of acetyltropic acid by countercurrent chromatography and reverse phase liquid chromatography. K E Y W O R D Sacetyltropic acid, countercurrent chromatography, cyclodextrins, enantioseparation, liquid chromatography J Sep Sci 2020;43:681-688.

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Expression of Brugmansia candida Hyoscyamine 6beta-Hydroxylase gene in Saccharomyces cerevisiae and its potential use as biocatalyst

Cited by 31 publications

References 27 publications

Functional identification of hyoscyamine 6β-hydroxylase from Anisodus acutangulus and overproduction of scopolamine in genetically-engineered Escherichia coli

Functional identification of hyoscyamine 6β-hydroxylase from Anisodus acutangulus and overproduction of scopolamine in genetically-engineered Escherichia coli

Anisodamine Production from Natural Sources: Seedlings and Hairy Root Cultures of Argentinean and ColombianBrugmansia candidaPlants

Enantioseparation of acetyltropic acid by countercurrent chromatography with sulfobutyl ether‐β‐cyclodextrin as chiral selector

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