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  • 1
    ISSN: 1573-9368
    Keywords: Nicotiana tabacum ; transformed root cultures ; metabolic engineering ; lysine decarboxylase ; protein targeting ; Rubisco transit peptide ; anabasine ; cadaverine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The gene of a bacterial lysine decarboxylase (ldc) fused to arbcS transit peptide coding sequence (tp), and under the control of the CaMV 35S promoter, was expressed in hairy root cultures ofNicotiana tabacum. The fusion of theldc to the targeting signal sequence improved the performance of the bacterial gene in the plant cells in many respects. Nearly all transgenic hairy root cultures harbouring the35S-tp-ldc gene contained distinctly higher lysine decarboxylase activity (from 1.5 to 30 pkat LDC per mg protein) than those which had been transformed with constructs in which the gene had been directly cloned behind the CaMV 35S promoter. The higher enzyme activity led to the accumulation of up to 0.7% cadaverine on a dry mass basis. In addition, part of the cadaverine pool was used for increased biosynthesis of anabasine, an alkaloid which was hardly detectable in control cultures. The best line contained anabasine levels of 0.5% dry mass, which could further be enhanced by feeding of lysine.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1573-5044
    Keywords: β-carboline alkaloids ; genetic manipulation ; hairy root culture ; Peganum harmala ; serotonin ; suspension culture
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract This article reviews our present knowledge on the formation of tryptophan derived secondary metabolites in tissue cultures of Peganum harmala. With the presence of β-carboline alkaloids and serotonin, P. harmala contains two rather simple, interrelated biosynthetic pathways. The long term disadvantage of low and unstable productivity of P. harmala suspension culture has recently been overcome by establishing highly productive hairy root cultures. The first β-carboline alkaloid biosynthetic enzymes, specific for the O-methylation of harmalol and harmol as well as for the oxidation of harmaline to harmine, have been detected in these cultures, and they should thus provide a suitable source for studying the yet unknown initial two enzymatic steps of β-carboline alkaloid biosynthesis. Seedlings of P. harmala have also been successfully transformed with constructed strains of Agrobacterium, as demonstrated by the overexpression of a tryptophan decarboxylase gene from Catharanthus roseus in cultures of P. harmala. In such transgenic cultures a large overproduction of serotonin was observed. The relative simplicity of these pathways and the rather easy handling of the cultures could make P. harmala a useful and attractive model system for studying the interaction, regulation and manipulation of secondary pathways in cultured cells.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1573-6776
    Source: Springer Online Journal Archives 1860-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract OneNicotiana glauca line with distinctly enhanced levels of lysine decarboxylase (LDC) activity and of cadaverine was detected among 54 hairy root cultures of different tobacco species, transformed with the binary vector pLX222 carrying a bacteial lysine decarboxylase gene directed by the 35S-promoter of CaMV. Anabasine levels of this line were nearly doubled in comparison to control lines transformed with the gus-gene instead of the ldc-gene.
    Type of Medium: Electronic Resource
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