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  • 1
    Electronic Resource
    Electronic Resource
    s.l. : American Chemical Society
    Journal of natural products 56 (1993), S. 1294-1303 
    ISSN: 1520-6025
    Source: ACS Legacy Archives
    Topics: Chemistry and Pharmacology
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    s.l. : American Chemical Society
    Journal of natural products 56 (1993), S. 1304-1312 
    ISSN: 1520-6025
    Source: ACS Legacy Archives
    Topics: Chemistry and Pharmacology
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: A dimethylallyl diphosphate:4-hydroxyphenylpyruvate dimethylallyl transferase from the novobiocin producer Streptomyces spheroides DSM 40292 was identified, partially purified and characterized. The enzyme was soluble, and specific for 4-hydroxyphenylpyruvate and dimethylallyl diphosphate as substrates. It is most likely involved in the biosynthesis of the prenylated 4-hydroxybenzoate moiety of the aminocoumarin antibiotic novobiocin.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 0340-1855
    Keywords: Schlüsselwörter Arthrose –¶Weidenrinde –Salix– WOMAC ; Key words Osteoarthritis –¶willow bark –Salix– WOMAC
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Summary Objective: To assess the clinical efficacy of a chemically standardized willow bark extract in the treatment of osteoarthritis. Methods: Willow bark extract, in a dose corresponding to 240mg salicin/day, was compared to placebo in a 2-week, double-blind, randomized controlled trial. The primary outcome measure was the pain dimension of the WOMAC Osteoarthritis Index. Secondary outcome measures included the stiffness and physical function dimensions of the WOMAC, daily visual analogue scales (VAS) on pain and physical function, and final overall assessment by both patients and investigators. Results: 78 patients (39 willow bark extract, 39 placebo) participated in the trial. A statistically significant difference between active treatment and placebo group was observed in the WOMAC pain dimension (d=6.5mm, 95%¶C.I.=0.2–12.7mm, p=0.047); the WOMAC pain score was reduced by 14% from baseline level after two weeks of active treatment, compared to an increase of 2% in the placebo group. Patient diary VAS confirmed this result, and likewise the final overall assessments showed superiority of willow bark extract over placebo ¶(patients assessment, p=0.0002; investigators assessment, p=0.0073). Conclusion: Willow bark extract shows a moderate analgesic effect in osteoarthritis.
    Notes: Zusammenfassung Studienziel: Untersuchung der analgetischen Wirksamkeit eines chemisch standardisierten Weidenrindenextraktes bei der Behandlung von Arthrose. Methoden: Weidenrindenextrakt, in einer Dosis entsprechend 240mg Salicin pro Tag, wurde in einer¶2-wöchigen, doppelblinden, randomisierten Studie gegen Placebo geprüft. Hauptzielparameter war die Schmerzdimension des WOMAC-Arthrose-Indexes. Als Nebenzielparameter dienten die WOMAC-Teilscores zu Steifigkeit und Funktionsfähigkeit, tägliche visuelle Analogskalen (VAS) zu Schmerz und Bewegungseinschränkung und das abschließende Gesamturteil sowohl des Patienten als auch des Arztes. Ergebnisse: 78 Patienten (39 Weidenrindenextrakt, 39 Placebo) nahmen an der Studie teil. Ein statistisch signifikanter Unterschied zwischen Verum- und Placebogruppe wurde für die WOMAC-Schmerzdimension beobachtet (d=6,5mm, 95% C.I.=0,2–12,7mm, p=0,047); während der Behandlung verringerte sich der WOMAC-Schmerzscore gegenüber dem Ausgangswert um 14% in der Verumgruppe, verglichen mit einem Anstieg von 2% in der Placebogruppe. Dieses Ergebnis wurde durch die VAS der Patiententagebücher bestätigt. Die abschließenden Gesamturteile zeigten eine deutliche Überlegenheit des Weidenrindenextraktes gegenüber Placebo (Patientenurteil, p=0,0002; Arzturteil, p=0,0073). Schlussfolgerung: Weidenrindenextrakt zeigt eine moderate analgetische Wirksamkeit bei Arthrosepatienten.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1432-2048
    Keywords: Key words: Geranyltransferase ; Lithospermum ; Methyl jasmonate ; Prenyltransferase ; Shikonin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract. Geranyldiphosphate:4-hydroxybenzoate 3-geranyltransferase is a regulatory enzyme in the biosynthesis of shikonin, a phytoalexin and pharmaceutical produced by cell cultures of Lithospermum erythrorhizon Sieb. et Zucc.. In Linsmaier-Skoog medium, the activity of this enzyme could be enhanced more than 200-fold by addition of methyl jasmonate, and this culture material was used for the solubilization and purification of the enzyme. Of various detergents examined, digitonin was the most suitable for the solubilization of the enzyme. The solubilized enzyme was purified 800-fold by chromatography over diethylaminoethyl (DEAE)-Sephacel, Heparin-Sepharose, Reactive Green 19-Agarose, and Cholic Acid-Agarose. The purified enzyme required magnesium ions as cofactor and was highly specific for geranyldiphosphate (GPP) and 4-hydroxybenzoate (4HB) as substrates. The K m values for 4HB and GPP were calculated by the method of Lineweaver and Burk as 18.4 μM and 13.8 μM, respectively.
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1432-203X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract o-Succinylbenzoic acid (OSB) is an intermediate in the biosynthesis of shikimatederived anthraquinones. The cell free activation of o-succinylbenzoic acid in extracts of anthraquinone producing cells of Galium mollugo L. is demonstrated for the first time. This activation depends on the presence of ATP, coenzyme A and Mg2+. The o-succinylbenzoic acid coenzyme A ester was identified by converting it to 1,4-dihydroxy-2-naphthoic acid by a bacterial enzyme, viz. naphthoatesynthase. It is thus demonstrated that the o-succinylbenzoic acid coenzyme A ester derived from bacteria and from Galium mollugo cells are identical.
    Type of Medium: Electronic Resource
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  • 7
    ISSN: 1432-2048
    Keywords: Key words: 3-Hydroxy-3-methylglutaryl-coenzyme A reductase ; Isoprenoid biosynthesis ; Light regulation ; Lithospermum ; Mevalonate pathway ; Shikonin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract. The carbon skeleton of the naphthoquinone pigment shikonin, which is produced in Lithospermum erythrorhizon Sieb. et Zucc. cell-suspension cultures, is partly derived from the isoprenoid biosynthetic pathway. The requirement of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMGR, EC 1.1.1.34), a key enzyme of the mevalonate route to isoprenoids, for shikonin synthesis was investigated. Conserved regions of sequences from plant HMGR genes were used to design polymerase chain reaction (PCR) primers for the cloning of a cDNA fragment from L. erythrorhizon. The resulting 443-bp clone was used as a probe for Northern analyses and hybridized to an mRNA of approx. 2.5 kb. Under shikonin-producing conditions, microsomal HMGR enzyme activity as well as mRNA level closely correlated with the accumulation of shikonin derivatives. White light, which inhibits shikonin formation, was shown to strongly suppress HMGR gene expression. The results presented here indicate that HMGR plays a significant role in the regulation of shikonin biosynthesis and that the control appears to act at the transcriptional level.
    Type of Medium: Electronic Resource
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  • 8
    ISSN: 1432-203X
    Keywords: Abbreviations: ADH = alcohol dehydrogenase; DTT = dithiothreitol; PMSF = phenylmethylsulfonyl fluoride; PVPP = polyvinylpolypyrrolidone; IAA = indole-3-acetic acid; TFA = trifluoroacetic acid
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract. An NAD-dependent alcohol dehydrogenase has been purified to apparent homogeneity from cell suspension cultures of Lithospermum erythrorhizon Sieb. et Zucc. (Boraginaceae), using protamine sulphate and ammonium sulphate precipitation and chromatography on DEAE-Sephacel, Superdex 200, hydroxyapatite and HiTrap blue. The enzyme is a homodimer with a Mr of ca. 77,000. Each subunit with a Mr of 40,000 contains two zinc atoms. Its isoelectric point was found at pH 5.0. The best alcohol substrate of the enzyme is ethanol. The pH optimum for ethanol oxidation is at pH 8.7 and for acetaldehyde reduction at pH 4.6. The Michaelis constants for ethanol and NAD are 2.49 and 0.05 (pH 8.7), and for acetaldehyde and NADH 2.2 and 0.078 mM (pH 4.6), respectively. Partial amino acid sequences of the purified enzyme showed high homology to alcohol dehydrogenases from other plants.
    Type of Medium: Electronic Resource
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  • 9
    ISSN: 1432-203X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract An NAD-dependent alcohol dehydrogenase has been purified to apparent homogeneity from cell suspension cultures of Lithospermum erythrorhizon Sieb. et Zucc. (Boraginaceae), using protamine sulphate and ammonium sulphate precipitation and chromatography on DEAE-Sephacel, Superdex 200, hydroxyapatite and HiTrap blue. The enzyme is a homodimer with a Mr of ca. 77,000. Each subunit with a Mr of 40,000 contains two zinc atoms. Its isoelectric point was found at pH 5.0. The best alcohol substrate of the enzyme is ethanol. The pH optimum for ethanol oxidation is at pH 8.7 and for acetaldehyde reduction at pH 4.6. The Michaelis constants for ethanol and NAD are 2.49 and 0.05 (pH 8.7), and for acetaldehyde and NADH 2.2 and 0.078 mM (pH 4.6), respectively. Partial amino acid sequences of the purified enzyme showed high homology to alcohol dehydrogenases from other plants.
    Type of Medium: Electronic Resource
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  • 10
    ISSN: 1432-2048
    Keywords: Key words: Cytochrome P-450 – Geranylhydroquinone 3′′-hydroxylase – 3′′-Hydroxy-geranylhydroquinone –Lithospermum– Naphthoquinone – Shikonin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract. Geranylhydroquinone 3′′-hydroxylase, which is likely to be involved in shikonin and dihydroechinofuran biosynthesis, was identified in cell suspension cultures of Lithospermum erythrorhizon Sieb. et Zucc. (Boraginaceae). The enzyme hydroxylates the isoprenoid side chain of geranylhydroquinone (GHQ), a known precursor of shikonin. Proton/proton correlation spectroscopic and proton/proton long-range correlation spectroscopic studies confirmed that hydroxylation takes place specifically at position 3′′, i.e. at the methyl group involved in the cyclization reaction. The enzyme is membrane-bound and was found in the microsomal fraction. It requires NADPH and molecular oxygen as cofactors, and is inhibited by cytochrome P-450 inhibitors such as cytochrome c and CO. The inhibitory effect of CO is reversed by illumination. These data suggest that the enzyme is a cytochrome P-450-dependent monooxygenase. The optimum pH of GHQ 3′′-hydroxylase is 7.4, and the apparent K m value for GHQ is 1.5 μM. The reaction velocity obtained with 3-geranyl-4-hydroxybenzoic acid was more than 100 times lower than that obtained with geranylhydroquinone.
    Type of Medium: Electronic Resource
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