PubMed Health⌕ Search
Home/Archive/PubMed

PubMed · 11587340

Automated multiple development thin layer chromatography for calystegines and their biosynthetic precursors.

Y Scholl·N Asano·B Dräger
View original source ↗Export metadata ↓

Abstract

Automated multiple development thin layer chromatography (AMD-TLC) was used for separation of calystegines, a class of nortropane alkaloids, and of precursors of their biosynthesis. The calystegines differ in the number of hydroxyl groups at the nortropane ring system and in the substitution pattern. A combination of TLC methods allows the separation of the individual calystegines and separation of possible precursors of the biosynthesis. Solvent combination, development time, the number of development steps, the drying time between each run and the preconditioning parameters of the silica gel TLC plates were optimised. Limits of detection were evaluated for several detection reagents.

Explore related subjects

AlkaloidsChromatography, Thin LayerPlant ExtractsTropanesJournal ArticleResearch Support, Non-U.S. Gov't

Keep this discovery

Explore connections, maps & timelines

BibTeXRIS

Y Scholl, N Asano, B Dräger. 2001-09-14. Automated multiple development thin layer chromatography for calystegines and their biosynthetic precursors.. https://doi.org/10.1016/s0021-9673(01)01128-1

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related citations

PubMed2007

Enantioselective total synthesis of (+)-gliocladin C.

The first total synthesis of gliocladin C, a fungal-derived marine alkaloid containing a rare trioxopiperazine fragment, is reported. This asymmetric synthesis establishes the absolute configuration of this structurally novel natural product. [reaction: see text].

Alkaloids↗
PubMed2007

Two novel glycosidic triterpene alkaloids from the stem barks of Machilus yaoshansis.

[structure: see text] Two unusual glycosidic triterpene alkaloids, machilaminosides A (1) and B (2), have been isolated from the stem barks of Machilus yaoshansis. Their structures were elucidated by detailed spectroscopic analysis. A possible biogenetic origin of 1 and 2 mediated by the coupling of 2-O-beta-D-glucopyranosyl-cucurbitacin I, respectively, with urea and adenosine was postulated. 1 and 2 showed nonselective cytotoxic activities against several human cancer cell lines as well as TNF-alpha secretion inhibitory activities.

Alkaloids↗
PubMed2007

Functional analysis of norcoclaurine synthase in Coptis japonica.

(S)-Norcoclaurine is the entry compound in benzylisoquinoline alkaloid biosynthesis and is produced by the condensation of dopamine and 4-hydroxyphenylacetaldehyde (4-HPAA) by norcoclaurine synthase (NCS) (EC 4.2.1.78). Although cDNA of the pathogenesis-related (PR) 10 family, the translation product of which catalyzes NCS reaction, has been isolated from Thalictrum flavum, its detailed enzymological properties have not yet been characterized. We report here that a distinct cDNA isolated from Coptis japonica (CjNCS1) also catalyzed NCS reaction as well as a PR10 homologue of C. japonica (CjPR10A). Both recombinant proteins stereo-specifically produced (S)-norcoclaurine by the condensation of dopamine and 4-HPAA. Because a CjNCS1 cDNA that encoded 352 amino acids showed sequence similarity to 2-oxoglutarate-dependent dioxygenases of plant origin, we characterized the properties of the native enzyme. Sequence analysis indicated that CjNCS1 only contained a Fe(2+)-binding site and lacked the 2-oxoglutarate-binding domain. In fact, NCS reaction of native NCS isolated from cultured C. japonica cells did not depend on 2-oxoglutarate or oxygen, but did require ferrous ion. On the other hand, CjPR10A showed no specific motif. The addition of o-phenanthroline inhibited NCS reaction of both native NCS and recombinant CjNCS1, but not that of CjPR10A. In addition, native NCS and recombinant CjNCS1 accepted phenylacetaldehyde and 3,4-dihydroxyphenylacetaldehyde, as well as 4-HPAA, for condensation with dopamine, whereas recombinant CjPR10A could use 4-hydroxyphenylpyruvate and pyruvate in addition to the above aldehydes. These results suggested that CjNCS1 is the major NCS in C. japonica, whereas native NCS extracted from cultured C. japonica cells was more active and formed a larger complex compared with recombinant CjNCS1.

Alkaloids↗