Cloning and Functional Expression in Escherichia coli of a cDNA Encoding Cardenolide 16′-O-Glucohydrolase from Digitalis lanata Ehrh.

Framm, Johann J.; Peterson, Angela; Thoeringer, Carola; Pangert, Antje; Hornung, Ellen; Feußner, Ivo; Luckner, Martin; Lindemann, Peter

doi:10.1093/pcp/pcd060

Cited by 11 publications

(5 citation statements)

References 32 publications

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“…Such modifications are supposed to be determinant in the intracellular and long‐distance transport of cardenolides. CGH may also play an important part in the defense against predators (Framm et al. , 2000).…”

Section: Resultsmentioning

confidence: 99%

“…Therefore, qualitative variations in secondary metabolites may play a role in plant defense which is as relevant as that of the quantitative changes observed in response to herbivore attacks. Interestingly, the CGH enzyme deglucosylating primary cardenolides in Digitalis may also be an important factor in the defense of these species against predators, because the secondary cardenolides formed are better resorbed and more toxic than their primary counterparts (Framm et al. , 2000).…”

Section: Discussionmentioning

confidence: 99%

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Digitalis purpurea P5βR2, encoding steroid 5β‐reductase, is a novel defense‐related gene involved in cardenolide biosynthesis

et al. 2009

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Summary The stereospecific 5β‐reduction of progesterone is a required step for cardiac glycoside biosynthesis in foxglove plants. Recently, we have isolated the gene P5βR, and here we investigate the function and regulation of P5βR2, a new progesterone 5β‐reductase gene from Digitalis purpurea. P5βR2 cDNA was isolated from a D. purpurea cDNA library and further characterized at the biochemical, structural and physiological levels. Like P5βR, P5βR2 catalyzes the 5β‐reduction of the Δ4 double bond of several steroids and is present in all plant organs. Under stress conditions or on treatment with chemical elicitors, P5βR expression does not vary, whereas P5βR2 is highly responsive. P5βR2 expression is regulated by ethylene and hydrogen peroxide. The correlation between P5βR2 expression and cardenolide formation demonstrates the key role of this gene in cardenolide biosynthesis, and therefore in the chemical defense of foxglove plants.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Digitalis purpurea P5βR2, encoding steroid 5β‐reductase, is a novel defense‐related gene involved in cardenolide biosynthesis

et al. 2009

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“…Nucleotide primers were deduced from these peptide fragments and a cgh1 cDNA was synthesized. After genetic transformation, a recombinant CGH I protein was produced in Escherichia coli [143]. Cotyledon explants of Cucumis sativus were genetically transformed using Agrobacterium rhizogenes harbouring a D. lanata cgh1 cDNA, and hairy roots formed at infected explants showed CGH I activity [144].…”

Section: Biosynthesis Of Cardenolidesmentioning

confidence: 99%

The Foxgloves (Digitalis) Revisited

Kreis

2017

Planta Med

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This review provides a renewed look at the genus Digitalis. Emphasis will be put on those issues that attracted the most attention or even went through paradigmatic changes since the turn of the millennium. PubMed and Google Scholar were used (“Digitalis” and “Foxglove” were the key words) to identify research from 2000 till 2017 containing data relevant enough to be presented here. Intriguing new results emerged from studies related to the phylogeny and taxonomy of the genus as well as to the biosynthesis and potential medicinal uses of the key active compounds, the cardiac glycosides. Several Eastern and Western Foxgloves were studied with respect to their propagation in vitro. In this context, molecular biology tools were applied and phytochemical analyses were conducted. Structure elucidation and analytical methods, which have experienced less exciting progress, will not be considered here in great detail.

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“…Phylogenetic analysis of the ␤-primeverosidase with various glycosyl hydrolases in this family is shown in Figure 4. The ␤-primeverosidase was grouped into the cluster of plant ␤-glucosidases, which act on various glycosides such as alkaloidal glucoside (strictosidine and raucaffricine; Geerlings et al, 2000;Warzecha et al, 2000), cardenolide glycoside (Framm et al, 2000), and in- doxyl glucoside (Minami et al, 1999). The ␤-primeverosidase was only loosely clustered with amygdalin hydrolase despite the high similarity between them because amygdalin hydrolase showed higher identities to prunasin hydrolase and linamarases and was grouped into the cluster of cyanogenic ␤-glucosidases.…”

Section: Characterization Of the Amino Acid Sequence Of ␤-Primeverosimentioning

confidence: 99%

Cloning of β-Primeverosidase from Tea Leaves, a Key Enzyme in Tea Aroma Formation

et al. 2002

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A ␤-primeverosidase from tea (Camellia sinensis) plants is a unique disaccharide-specific glycosidase, which hydrolyzes aroma precursors of ␤-primeverosides (6-O-␤-d-xylopyranosyl-␤-d-glucopyranosides) to liberate various aroma compounds, and the enzyme is deeply concerned with the floral aroma formation in oolong tea and black tea during the manufacturing process. The ␤-primeverosidase was purified from fresh leaves of a cultivar for green tea (C. sinensis var sinensis cv Yabukita), and its partial amino acid sequences were determined. The ␤-primeverosidase cDNA has been isolated from a cDNA library of cv Yabukita using degenerate oligonucleotide primers. The cDNA insert encodes a polypeptide consisting of an N-terminal signal peptide of 28 amino acid residues and a 479-amino acid mature protein. The ␤-primeverosidase protein sequence was 50% to 60% identical to ␤-glucosidases from various plants and was classified in a family 1 glycosyl hydrolase. The mature form of the ␤-primeverosidase expressed in Escherichia coli was able to hydrolyze ␤-primeverosides to liberate a primeverose unit and aglycons, but did not act on 2-phenylethyl ␤-d-glucopyranoside. These results indicate that the ␤-primeverosidase selectively recognizes the ␤-primeverosides as substrates and specifically hydrolyzes the ␤-glycosidic bond between the disaccharide and the aglycons. The stereochemistry for enzymatic hydrolysis of 2-phenylethyl ␤-primeveroside by the ␤-primeverosidase was followed by 1 H-nuclear magnetic resonance spectroscopy, revealing that the enzyme hydrolyzes the ␤-primeveroside by a retaining mechanism. The roles of the ␤-primeverosidase in the defense mechanism in tea plants and the floral aroma formation during tea manufacturing process are also discussed.

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Cloning and Functional Expression in Escherichia coli of a cDNA Encoding Cardenolide 16′-O-Glucohydrolase from Digitalis lanata Ehrh.

Cited by 11 publications

References 32 publications

Digitalis purpurea P5βR2, encoding steroid 5β‐reductase, is a novel defense‐related gene involved in cardenolide biosynthesis

Digitalis purpurea P5βR2, encoding steroid 5β‐reductase, is a novel defense‐related gene involved in cardenolide biosynthesis

The Foxgloves (Digitalis) Revisited

Cloning of β-Primeverosidase from Tea Leaves, a Key Enzyme in Tea Aroma Formation

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