The tropane alkaloid anisodamine (2) is obtained by 6β-hydroxylation of hyoscyamine (1). The application of this alkaloid in medicine is gaining attention due to the wide range of therapeutic applications described in addition to its anticholinergic activity. In this work, the production of anisodamine (2) by in vitro cultures of Brugmansia candida (Argentinean and Colombian samples) was studied. This alkaloid was estimated in different organs of in vitro-germinated seedlings as well as in hairy roots obtained from seedlings from both sources. Colombian roots exhibited the highest content of tropane alkaloids, with anisodamine (2) being the main alkaloid measured. In the leaves, the main alkaloid was scopolamine (3) and no significant differences were observed between Argentinean and Colombian leaves. The tropane alkaloid content in Argentinean hairy roots was significantly higher than in Colombian ones. Also, in the Argentinean samples the main alkaloid detected was anisodamine (2). Argentinean and Colombian B. candida seedlings and hairy roots appear to be a promising system for the production of anisodamine (2).
3
Palazón J, Moyano E, Cusidó R M, Bonfill M, Oksman-Caldentey K-M, Pinol M T.
Alkaloid production in Duboisia hybrid hairy roots and plants overexpressing the h6h gene.
Plant Sci.
2003;
165
1289-1295
5
Zhang L, Ding R, Chai Y, Bonfill M, Moyano E, Oksman-Caldentey K M, Xu T, Pi Y, Wang Z, Zhang H, Kai G, Liao Z, Sun X, Tang K.
Engineering tropane biosynthetic pathway in Hyoscyamus niger hairy root cultures.
Proc Natl Acad Sci USA.
2004;
101
6786-6791
7 Wang T N, Yang H J, Gu-Ling L, Li J Y, Zheng X X. Advanced measurement and quantitative appraise of anisodamine on calcium triggered in cardiac myocyte. Engineering in Medicine and Biology 27th Annual Conference, Shanghai, China; 2005.
9
Wang H, Lu Y, Chen H Z.
Differentiating effects of anisodamine on cognitive amelioration and peripheral muscarinic side effects induced by pilocarpine in mice.
Neurosci Lett.
2003;
344
173-176
13
Yang L M, Xie Y F, Chen H Z, Lu Y.
Diastereomeric and enantiomeric high-performance liquid chromatographic separation of synthetic anisodamine.
J Pharm Biomed Anal.
2007;
43
905-909
14
Lee K T, Yamakawa T, Kodama T, Shimomura K.
Effects of chemicals on alkaloid production by transformed roots of Belladonna.
Phytochemistry.
1998;
49
2343-2347
15
Li J, Chun Y, Jua H.
Simultaneous electrochemiluminiscence detection of anisodamine, atropine and scopolamine in Flos daturae by capillary electrophoresis using b-cyclodextrin as additive.
Electroanalysis.
2007;
19
1569-1574
16
Diwan R, Malpathak N.
Novel technique for scaling up of micropropagated Ruta graveolens shoots using liquid culture systems: a step towards commercialization.
New Biotechnol.
2008;
25
85-91
19
Hashimoto T, Hayashi A, Amano Y, Kohno J, Iwanari H, Usuda S, Yamada Y.
Hyoscyamine 6beta-hydroxylase, an enzyme involved in tropane alkaloid biosynthesis, is localized at the pericycle of the root.
J Biol Chem.
1991;
266
4648-4653
20
Zarate R, el Jaber-Vazdekis N, Medina B, Ravelo A G.
Tailoring tropane alkaloid accumulation in transgenic hairy roots of Atropa baetica by over-expressing the gene encoding hyoscyamine 6beta-hydroxylase.
Biotechnol Lett.
2006;
28
1271-1277
22
Cardillo A B, Giulietti A M, Marconi P L.
Analysis and sequencing of h6hmRNA, last enzyme in the tropane alkaloids pathway from anthers and hairy root cultures of Brugmansia candida (Solanaceae).
Electronic J Biotechnol.
2006;
9
195-198
23
Cardillo A B, Rodriguez Talou J, Giulietti A M.
Expression of Brugmansia candida hyoscyamine 6beta-hydroxylase gene in Saccharomyces cerevisiae and its potential use as biocatalyst.
Microb Cell Fact.
2008;
7
17
24
Kursinszki L, Hank H, Laszlo I, Szoke E.
Simultaneous analysis of hyoscyamine, scopolamine, 6beta-hydroxyhyoscyamine and apoatropine in Solanaceous hairy roots by reversed-phase high-performance liquid chromatography.
J Chromatogr A.
2005;
1091
32-39
25
Cheng K D, Zhu W H, Li X L, Meng C, Sun Z M, Yang D H.
Biotransformation of hyoscyamine by suspension cultures of Anisodus tanguticus.
Planta Med.
1987;
53
211-213
26
Zhang H M, Ou Z L, Yamamoto T.
Anisodamine inhibits shiga toxin type 2-mediated tumor necrosis factor-alpha production in vitro and in vivo.
Exp Biol Med (Maywood).
2001;
226
597-604
27
Pitta-Alvarez S I, Spollansky T C, Giulietti A M.
The influence of different biotic and abiotic elicitors on the production and profile of tropane alkaloids in hairy root cultures of Brugmansia candida.
Enzyme Microb Technol.
2000;
26
252-258
29
Hamill J D, Rounsley S, Spencer A, Todd G, Rhodes J C.
The use of the polymerase chain reaction in plant transformation studies.
Plant Cell Rep.
1991;
10
221-224