Biosynthesis of serotonin and β-carboline alkaloids in hairy root cultures of Peganum harmala

Berlin, Jochen; Rügenhagen, C.; Greidziak, N.; Кузовкина, И. Н.; Witte, Ludger; Wray, Victor

doi:10.1016/0031-9422(93)85453-x

Cited by 30 publications

(23 citation statements)

References 21 publications

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“…Control (not tdc transformed) cell suspensions and root cultures of P. harmala were derived similarly from infections with LBA4404 (pBDH5) or C58CI pRiA4 (pBDH5) or were from our culture collection, i.e. PHF (suspension line), and PH A4 or PH gus (root lines) (Berlin et al, , 1993. Growth and maintenance of suspension and root cultures were as described previously (Berlin et al, , 1993.…”

Section: Plant Cell Suspension and Hairy Root Culturesmentioning

confidence: 99%

“…PHF (suspension line), and PH A4 or PH gus (root lines) (Berlin et al, , 1993. Growth and maintenance of suspension and root cultures were as described previously (Berlin et al, , 1993.…”

Section: Plant Cell Suspension and Hairy Root Culturesmentioning

confidence: 99%

“…Consequently, serotonin, but not ~-carboline alkaloid, production was restored by selecting from a nonproducing cell suspension culture 4-methyltryptophantolerant cell lines showing reasonable TDC activity . These amino-acid-analogue-tolerant cell lines, with serotonin levels of 1 to 2% dry mass, have proved to be rather stable with respect to TDC activity and serotonin production (Berlin et al, 1993). Serotonin biosynthesis in P. harmala thus offers a unique system in which to test whether the rate-limiting step of a biosynthetic pathway of plant secondary metabolism can be overcome by introducing genetically a 'foreign' enzyme activity catalysing that reaction.…”

Section: Introductionmentioning

confidence: 99%

“…It was shown that the lack or loss of tryptophan decarboxylase (TDC) activity is apparently the reason why serotonin biosynthesis ceases in suspension cultures of P. harmala . Tryptamine supply seems also to be yield-limiting for serotonin biosynthesis in root cultures (Berlin et aL, 1993). In the case of (3-carboline alkaloids, additional, as yet unknown regulatory controls must have been turned off in non-producing suspension cultures .…”

Section: Introductionmentioning

confidence: 99%

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Increased production of serotonin by suspension and root cultures ofPeganum harmala transformed with a tryptophan decarboxylase cDNA clone fromCatharanthus roseus

Berlin

Rügenhagen

Dietze

et al. 1993

Transgenic Research

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Cell suspension and root cultures of Peganum harmala were established expressing a tryptophan decarboxylase cDNA clone from Catharanthus roseus under the control of the cauliflower mosaic virus (CaMV) 35S promoter and terminator sequences. The tryptophan decarboxylase activity of some of the transgenic lines was greatly enhanced (25-40 pkat/mg protein) as compared to control cultures (1-5 pkat per mg protein) and remained high during the growth cycle. While the levels of tryptamine, the product of the reaction catalysed by tryptophan decarboxylase, were unchanged in the transgenic lines, their serotonin contents were enhanced up to 10-fold, reaching levels of 1.5 to 2% dry mass. Thus, tryptamine produced by the engineered reaction was apparently immediately used for enhanced serotonin biosynthesis. The yields of serotonin in transgenic lines overexpressing tryptophan decarboxylase activity were further enhanced to 3-5% dry mass by feeding L-tryptophan, while no or only minor effects were seen when control cultures were fed. These data demonstrate that the production of a plant secondary metabolite can be enhanced greatly via genetic manipulation of the level of activity of the rate-limiting enzyme. The amounts of 13-carboline alkaloids, the other tryptamine-derived metabolites of P. harmala, in contrast, were not affected by the overproduction of tryptamine. The information needed for successfully predicting manipulations that enhance production of a secondary metabolite is discussed.

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Section: Plant Cell Suspension and Hairy Root Culturesmentioning

confidence: 99%

“…PHF (suspension line), and PH A4 or PH gus (root lines) (Berlin et al, , 1993. Growth and maintenance of suspension and root cultures were as described previously (Berlin et al, , 1993.…”

Section: Plant Cell Suspension and Hairy Root Culturesmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Increased production of serotonin by suspension and root cultures ofPeganum harmala transformed with a tryptophan decarboxylase cDNA clone fromCatharanthus roseus

Berlin

Rügenhagen

Dietze

et al. 1993

Transgenic Research

Self Cite

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show abstract

“…For example, hyoscyamine-6-B-hydroxylase, a key enzyme in the biosynthesis of the tropane alkaloids, has been characterized in root cultures of Hyoscyamus, 105 along with other enzymes in this pathway: tropinone reductase from D. stramonium, 106 [115][116][117] Other pathways currently under study are: thiophenes in C douglas ii, 118 bisbenzylisoquinoline alkaloids in Stephania cepharantha,119 thiarubrine A in Ambrosia artemisiijolia, 120 and serotonin and B-carbolide alkaloids in Peganum harmala. 121 In our laboratory, efforts are directed to the study of indole alkaloid biosynthesis in hairy root cultures of C roseus. We have purified tryptophan decarboxylase to homogenity, 122 and have partially purified strictosidine synthase (Munoz, Moreno-Valenzuela and Loyola-Vargas, unpublished data), 3-hydroxymethylglutaryl CoA reductase (Galaz and Loyola-Vargas, unpublished re-suIts), and geraniol lO-hydroxylase (Canto and Loyola-Vargas, unpublished data).…”

Section: Biochemical Studiesmentioning

confidence: 99%

Root Culture as a Source of Secondary Metabolites of Economic Importance

Loyola‐Vargas

Miranda-Ham

1995

Phytochemistry of Medicinal Plants

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Production of monoclonal antibodies by tobacco hairy roots

Wongsamuth

Doran

1997

Biotechnol. Bioeng.

147

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Hairy roots of tobacco (Nicotiana tabacum) were used to produce full‐length murine lgG1 monoclonal antibody. The presence of heavy (γ) and light (κ) chains and fully assembled antibody was verified by Western blot analysis of root extracts. Antibody levels in the biomass and medium were quantified by ELISA based on detection of γ‐κ complexes. Antibody produced by hairy roots was fully functional as demonstrated in bacterial aggregation assays which confirmed bivalent antigen‐binding capacity. Eight antibody‐producing hairy root clones retained their ability to produce mouse immunoglobulin over a period of 19 months after transformation with Agrobacterium rhizogenes. For hairy roots grown in Gamborg's B5 medium, the maximum level of assembled antibody after 21‐day culture in shake flasks was 18 mg L−1 or 1.8% total soluble protein; up to 14% of the antibody was secreted into the medium. Antibody production by transgenic hairy roots had a negligible effect on growth compared with hairy roots of wild‐type tobacco. Antibody accumulation was growth associated with constant specific accumulation rate at the beginning of the culture; however, degradation of antibody was significant after 14 days and the amount of assembled antibody declined. Unlike hybridoma cultures, the time course of antibody accumulation by hairy roots showed a distinctive maximum very soon after the end of exponential growth. Total antibody levels were increased by addition of nitrate, polyvinylpyrrolidone, or gelatin to the medium. Polyvinylpyrrolidone and gelatin also markedly improved extracellular antibody concentrations; with these treatments, up to 43% of the antibody present was secreted into the medium. Antibody production was tested using hairy roots grown in an air‐driven bioreactor. The intracellular antibody content after 30‐day bioreactor culture was similar to that measured in shake flasks; however, the final extracellular antibody level was 1.7 times higher than the maximum measured in shake flasks. © 1997 John Wiley & Sons, Inc. Biotechnol Bioeng 54: 401–415, 1997.

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Biosynthesis of serotonin and β-carboline alkaloids in hairy root cultures of Peganum harmala

Cited by 30 publications

References 21 publications

Increased production of serotonin by suspension and root cultures ofPeganum harmala transformed with a tryptophan decarboxylase cDNA clone fromCatharanthus roseus

Increased production of serotonin by suspension and root cultures ofPeganum harmala transformed with a tryptophan decarboxylase cDNA clone fromCatharanthus roseus

Root Culture as a Source of Secondary Metabolites of Economic Importance

Production of monoclonal antibodies by tobacco hairy roots

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