Metabolic engineering of the indole pathway in Catharanthus roseus hairy roots and increased accumulation of tryptamine and serpentine

Erik H. Hughes, Seung Beom Hong, Sue Gibson, Jacqueline V. Shanks, Ka Yiu San

Research output: Contribution to journalArticle

89 Citations (Scopus)

Abstract

Transgenic hairy roots of Catharanthus roseus were established with glucocorticoid inducible tryptophan decarboxylase (TDC) expression alone or in combination with inducible expression of a feedback-resistant anthranilate synthase alpha subunit (ASα) from Arabidopsis. Northern blot analysis confirmed transcription of the anthranilate synthase gene upon induction in the double line (TDC+ASα) and in vitro enzyme assays confirmed increased resistance to feedback inhibition by tryptophan. In TDC enzyme assays, increases of 48% and 87% in the TDC and double lines, respectively, were noted. Although the TDC line showed no significant increase in tryptamine levels on induction, induction of the double line resulted in increases in tryptamine levels of as much as six-fold for a 3 day late exponential induction. Downstream effects on alkaloids were noted in the TDC line where serpentine specific yields increased as much as 129% on induction. No effects on measured alkaloids were noted in the double line, but the two clones have very different basal alkaloid biosynthetic capacities. Within this study, the engineering of the indole pathway in C. roseus hairy roots is reported, and the role of the indole pathway in alkaloid biosynthesis explored.

Original languageEnglish (US)
Pages (from-to)268-276
Number of pages9
JournalMetabolic Engineering
Volume6
Issue number4
DOIs
StatePublished - Oct 1 2004

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Catharanthus
Aromatic-L-Amino-Acid Decarboxylases
Metabolic engineering
Metabolic Engineering
Alkaloids
Assays
Enzymes
Enzyme Assays
Feedback
Enzyme inhibition
Biosynthesis
Anthranilate Synthase
Transcription
Genes
Tryptophan
Northern Blotting
Glucocorticoids
tryptamine
serpentine (alkaloid)
indole

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Metabolic engineering of the indole pathway in Catharanthus roseus hairy roots and increased accumulation of tryptamine and serpentine. / Hughes, Erik H.; Hong, Seung Beom; Gibson, Sue; Shanks, Jacqueline V.; San, Ka Yiu.

In: Metabolic Engineering, Vol. 6, No. 4, 01.10.2004, p. 268-276.

Research output: Contribution to journalArticle

Hughes, Erik H. ; Hong, Seung Beom ; Gibson, Sue ; Shanks, Jacqueline V. ; San, Ka Yiu. / Metabolic engineering of the indole pathway in Catharanthus roseus hairy roots and increased accumulation of tryptamine and serpentine. In: Metabolic Engineering. 2004 ; Vol. 6, No. 4. pp. 268-276.
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