Stable vindoline production in transformed cell cultures of Catharanthus roseus

Barry R. O'Keefe; Gail B. Mahady; Joell J. Gills; Christopher W.W. Beecher; Alex B. Schilling

doi:10.1021/np960703n

Stable vindoline production in transformed cell cultures of Catharanthus roseus

Barry R. O'Keefe, Gail B. Mahady, Joell J. Gills, Christopher W.W. Beecher, Alex B. Schilling

Research output: Contribution to journal › Article › peer-review

Abstract

Catharanthus roseus L. (G. Don) leaf disks were transformed by cocultivation with seven strains of Agrobacterium tumefaciens and Agrobacterium rhizogenes. Four distinct transformed cell lines were obtained with A. tumefaciens strains A281, BO542, and A. rhizogenes strain K599, and selected on hormone-free medium. Transformation of the cell lines was confirmed by Southern blot analysis, and preliminary data on the alkaloid profiles was determined by 2D TLC and HPLC/MS. Stable vindoline production was demonstrated in the undifferentiated suspension cell lines strains, CR- BO542, CR-K599, and strain CR-A281S (shooty teratomas), by HPLC/MS analysis of the alkaloid fractions. Active vindoline biosynthesis in the suspension- cell lines was further confirmed by measuring the activity of acetyl- coenzyme-A-deacetylvindoline-O-acetyltransferase, one of the final enzymes in the biosynthetic pathway of vindoline, in cell-free extracts.

Original language	English (US)
Pages (from-to)	261-264
Number of pages	4
Journal	Journal of Natural Products
Volume	60
Issue number	3
DOIs	https://doi.org/10.1021/np960703n
State	Published - Apr 15 1997
Externally published	Yes

ASJC Scopus subject areas

Analytical Chemistry
Molecular Medicine
Pharmacology
Pharmaceutical Science
Drug Discovery
Complementary and alternative medicine
Organic Chemistry

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title = "Stable vindoline production in transformed cell cultures of Catharanthus roseus",

abstract = "Catharanthus roseus L. (G. Don) leaf disks were transformed by cocultivation with seven strains of Agrobacterium tumefaciens and Agrobacterium rhizogenes. Four distinct transformed cell lines were obtained with A. tumefaciens strains A281, BO542, and A. rhizogenes strain K599, and selected on hormone-free medium. Transformation of the cell lines was confirmed by Southern blot analysis, and preliminary data on the alkaloid profiles was determined by 2D TLC and HPLC/MS. Stable vindoline production was demonstrated in the undifferentiated suspension cell lines strains, CR- BO542, CR-K599, and strain CR-A281S (shooty teratomas), by HPLC/MS analysis of the alkaloid fractions. Active vindoline biosynthesis in the suspension- cell lines was further confirmed by measuring the activity of acetyl- coenzyme-A-deacetylvindoline-O-acetyltransferase, one of the final enzymes in the biosynthetic pathway of vindoline, in cell-free extracts.",

author = "O'Keefe, {Barry R.} and Mahady, {Gail B.} and Gills, {Joell J.} and Beecher, {Christopher W.W.} and Schilling, {Alex B.}",

year = "1997",

month = apr,

day = "15",

doi = "10.1021/np960703n",

language = "English (US)",

volume = "60",

pages = "261--264",

journal = "Journal of Natural Products",

issn = "0163-3864",

publisher = "American Chemical Society",

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TY - JOUR

T1 - Stable vindoline production in transformed cell cultures of Catharanthus roseus

AU - O'Keefe, Barry R.

AU - Mahady, Gail B.

AU - Gills, Joell J.

AU - Beecher, Christopher W.W.

AU - Schilling, Alex B.

PY - 1997/4/15

Y1 - 1997/4/15

N2 - Catharanthus roseus L. (G. Don) leaf disks were transformed by cocultivation with seven strains of Agrobacterium tumefaciens and Agrobacterium rhizogenes. Four distinct transformed cell lines were obtained with A. tumefaciens strains A281, BO542, and A. rhizogenes strain K599, and selected on hormone-free medium. Transformation of the cell lines was confirmed by Southern blot analysis, and preliminary data on the alkaloid profiles was determined by 2D TLC and HPLC/MS. Stable vindoline production was demonstrated in the undifferentiated suspension cell lines strains, CR- BO542, CR-K599, and strain CR-A281S (shooty teratomas), by HPLC/MS analysis of the alkaloid fractions. Active vindoline biosynthesis in the suspension- cell lines was further confirmed by measuring the activity of acetyl- coenzyme-A-deacetylvindoline-O-acetyltransferase, one of the final enzymes in the biosynthetic pathway of vindoline, in cell-free extracts.

AB - Catharanthus roseus L. (G. Don) leaf disks were transformed by cocultivation with seven strains of Agrobacterium tumefaciens and Agrobacterium rhizogenes. Four distinct transformed cell lines were obtained with A. tumefaciens strains A281, BO542, and A. rhizogenes strain K599, and selected on hormone-free medium. Transformation of the cell lines was confirmed by Southern blot analysis, and preliminary data on the alkaloid profiles was determined by 2D TLC and HPLC/MS. Stable vindoline production was demonstrated in the undifferentiated suspension cell lines strains, CR- BO542, CR-K599, and strain CR-A281S (shooty teratomas), by HPLC/MS analysis of the alkaloid fractions. Active vindoline biosynthesis in the suspension- cell lines was further confirmed by measuring the activity of acetyl- coenzyme-A-deacetylvindoline-O-acetyltransferase, one of the final enzymes in the biosynthetic pathway of vindoline, in cell-free extracts.

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