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  • 1
    ISSN: 1432-2048
    Keywords: Cell culture (coniferin accumulation) ; β-Coniferin ; Glucosidase ; Linum ; 5-Methoxypodophyllotoxin ; Peroxidase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Cell suspensions of Linum flavum L. contained large amounts (2 g·l−1) of the glucoside coniferin which was accumulated endogenously up to 12.4% on a dryweight basis. Callus material contained 5.6%, while in leaves of in-vitro-grown plantlets, the origin of the callus and suspension cultures, no coniferin could be detected. Leaf, callus and suspension material were compared for metabolite accumulation and associated enzyme activities. High coniferin contents corresponded with low 5-methoxypodophyllotoxin levels. A reciprocal relationship between β-glucosidase (E.C. 3.2.1.21) activity and coniferin accumulation was found. No relationship between peroxidase (E.C. 1.11.1.7) activity and coniferin accumulation or 5-methoxypodophyllotoxin could be demonstrated. Finally, a rapid and effective isolation procedure for coniferin was developed.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-203X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The phenylpropanoid derived lignan podophyllotoxin, occurring inPodophyllum species, is used as a starting compound for the chemical synthesis of the antitumour agents etoposide (VP-16-213) and teniposide (VM-26). At present, the availability of this lignan becomes increasingly limited. As an alternative source, cell cultures originating fromPodophyllum hexandrum Royle were initiated. Analysis of the cell extracts using different HPLC systems as well as TLC, indicated the presence of podophyllotoxin. After prepurification of the extracts by means of ITLC, the identity was confirmed by mass spectrometric analysis. Dark-grown cultures accumulated considerable higher amounts of podophyllotoxin in comparison with the light-grown cultures.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1573-739X
    Keywords: Cells, cultured ; Levodopa ; Mucuna pruriens ; Selection (genetics)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract Routinely grown cell suspension cultures ofMucuna pruriens L. (Fabaceae) were able to endogenously accumulate the anti-Parkinson drug L-dihydroxyphenylalanine (L-dopa) in the range between 0.2 and 2% on a dry weight (DW) basis. The green colour that developed in light-exposed cultures, appeared to be a suitable marker to select cells with an increased L-dopa biosynthesis and/or phenoloxidase activity. For this purpose, saccharose concentrations from 0 to 4% (w/v), and light intensities of 1,000 and 2,000 lux, were involved in the selection procedure. After 6 months, photomixotrophic callus cultures with a rapid growth and a high L-dopa content of 0.9% (DW) were obtained on 2% saccharose and under 1,000 lux. The cell suspensions, derived from these calli, accumulated up to 6% (DW) L-dopa, which was the highest stable content ever measured in cultures ofM. pruriens. An L-dopa yield of approximately 1.2 g/l was calculated after 6 days of growth. In contrast, compared with the standard-grown parent cell line, the phenoloxidase activity, and consequently the bioconversion capacity as measured after entrapment in calcium alginate, of these high-producing cultures was approximately threefold lower.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1573-5044
    Keywords: 2,4-d ; dopamine ; L-DOPA ; Mucuna pruriens ; plant cell culture ; salt
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The development of the L-DOPA-content of roots, stems and leaves ofMucuna pruriens during growth of the plants is described. Besides L-DOPA, the leaves, but not the stems and the roots, also contain the related catechol dopamine. The time course of dopamine accumulation is compared to that of L-DOPA. In cell suspension cultures ofM. pruriens dopamine can be detected as well. Its level is strongly increased by addition of the growth regulator 2,4-d to the medium, a condition that suppresses cell growth and L-DOPA-accumulation. Dopamine induction appears to be a specific metabolic effect of 2,4-d. Salt stress, as caused by the addition of NaCl, gives no induction of dopamine formation, whereas L-DOPA is released into the medium.
    Type of Medium: Electronic Resource
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