Metabolic effects of a bacterial lysine decarboxylase gene expressed in a hairy root culture ofNicotiana glauca

Fecker, Lothar F.; Hillebrandt, S.; Rügenhagen, C.; Herminghaus, S.; Landsmann, J.; Berlin, Jochen

doi:10.1007/bf01021054

Cited by 27 publications

(16 citation statements)

References 12 publications

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“…Roughly 80% of the lines transformed with the 35S-tp-ldc vector contained LDC activities between 2-12pkat per mg protein in the initial screenings. Thus, many more LDC positive lines were found in comparison to our transformation experiments using vectors in which the ldc gene was directly cloned behind the CaMV35S promoter (Fecker et al, 1992. In addition, the few LDC-positive lines of the previous transformations contained distinctly lower LDC activity.…”

Section: Screening Of Kanamycin-tolerant Root Culturesmentioning

confidence: 95%

“…Cadaverine and putrescine were determined as benzoyl derivatives (Redmond and Tseng, 1979) and the alkaloids (anabasine, nicotine and nornicotine) by GC or HPLC as described (Fecker et al, 1992.…”

Section: Determination Of Metabolitesmentioning

confidence: 99%

“…To verify this assumption, we tried to achieve overproduction of cadaverine by the expression of a bacterial lysine decarboxylase gene (Idc) of Hafnia alvei (Fecker et al, 1986) under the control of the CaMV35S promoter in hairy root cultures of tobacco. Indeed, we could show that cadaverine and anabasine levels were enhanced in some transgenic lines containing low, but significantly increased, lysine decarboxylase activity (LDC) (Fecker et al, 1992. Nevertheless, the metabolic potential of the LDC-positive lines for cadaverine and anabasine formation only became clearly visible when lysine was fed to the transgenic cultures .…”

Section: Introductionmentioning

confidence: 98%

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Improved metabolic action of a bacterial lysine decarboxylase gene in tobacco hairy root cultures by its fusion to arbcS transit peptide coding sequence

Herminghaus¹,

Tholl²,

Rügenhagen³

et al. 1996

Transgenic Research

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The gene of a bacterial lysine decarboxylase (ldc) fused to a rbcS transit peptide coding sequence (tp), and under the control of the CaMV 35S promoter, was expressed in hairy root cultures of Nicotiana tabacum. The fusion of the ldc 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 the 35S-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 be further be enhanced by feeding of lysine.

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Section: Screening Of Kanamycin-tolerant Root Culturesmentioning

confidence: 95%

Section: Determination Of Metabolitesmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 98%

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Improved metabolic action of a bacterial lysine decarboxylase gene in tobacco hairy root cultures by its fusion to arbcS transit peptide coding sequence

Herminghaus¹,

Tholl²,

Rügenhagen³

et al. 1996

Transgenic Research

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“…tion. A hairy root culture of Nicotiana glauca expressing a ldc gene originating from the gramnegative bacterium Hafnia alvei [6] had indeed distinctly higher levels of cadaverine as well as twice as much anabasine as a control culture [8].…”

Section: Introductionmentioning

confidence: 99%

“…In addition, we had faced the problem that the ldc gene used for plant transformation was not well translated into an active enzyme when cloned behind the CaMV 35S promoter. In the previous report it was shown that the establishment of LDC-positive root cultures was unefficient when a binary vector was used containing only one expression cassette [8]. Only 2 out of 54 kanamycin-resistant hairy root cultures carrying one copy of the bacterial Idc gene had given rise to LDC-positive lines.…”

Section: Introductionmentioning

confidence: 99%

Increased production of cadaverine and anabasine in hairy root cultures of Nicotiana tabacum expressing a bacterial lysine decarboxylase gene

Fecker¹,

Rügenhagen

Berlin

1993

Plant Mol Biol

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Several hairy root cultures of Nicotiana tabacum varieties, carrying two direct repeats of a bacterial lysine decarboxylase (ldc) gene controlled by the cauliflower mosaic virus (CaMV) 35S promoter expressed LDC activity up to 1 pkat/mg protein. Such activity was, for example, sufficient to increase cadaverine levels of the best line SR3/1-K1,2 from ca. 50 micrograms (control cultures) to about 700 micrograms/g dry mass. Some of the overproduced cadaverine of this line was used for the formation of anabasine, as shown by a 3-fold increase of this alkaloid. In transgenic lines with lower LDC activity the changes of cadaverine and anabasine levels were correspondingly lower and sometimes hardly distinguishable from controls. Feeding of lysine to root cultures, even to those with low LDC activity, greatly enhanced cadaverine and anabasine levels, while the amino acid had no or very little effect on controls and LDC-negative lines.

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Biochemistry and Physiology of Alkaloids and Betalains

Roberts

Strack

2018

Annual Plant Reviews Online

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The sections in this article are Introduction Nicotine and Tropane Alkaloids Pyrrolizidine Alkaloids Benzylisoquinoline Alkaloids Monoterpene Indole Alkaloids Ergot Alkaloids Acridone Alkaloid Biosynthesis Purine Alkaloids Taxol Betalains Conclusions Acknowledgements

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Metabolic effects of a bacterial lysine decarboxylase gene expressed in a hairy root culture ofNicotiana glauca

Cited by 27 publications

References 12 publications

Improved metabolic action of a bacterial lysine decarboxylase gene in tobacco hairy root cultures by its fusion to arbcS transit peptide coding sequence

Improved metabolic action of a bacterial lysine decarboxylase gene in tobacco hairy root cultures by its fusion to arbcS transit peptide coding sequence

Increased production of cadaverine and anabasine in hairy root cultures of Nicotiana tabacum expressing a bacterial lysine decarboxylase gene

Biochemistry and Physiology of Alkaloids and Betalains

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