RESUMO
The process of galanthamine and related alkaloids production by Leucojum aestivum shoot culture in a temporary immersion system was studied. It was established that temporary immersion approach is prospective for development of a biosynthetic process for obtaining valuable Amaryllidaceae alkaloids. Both immersion frequency and temperature had significant effect on biomass accumulation and the yields of galanthamine and related alkaloids. The maximal yield of galanthamine was achieved at the cultivation of L. aestivum shoot culture in temporary immersion RITA(®) system at immersion frequency 15 min flooding and 8 h stand-by periods, at 26 °C. Data on the relationships in the biological system "Nutrient medium-L. aestivum shoot culture-galanthamine" are presented as well.
Assuntos
Galantamina/biossíntese , Liliaceae/metabolismo , Brotos de Planta/metabolismo , Técnicas de Cultura de Células , Meios de Cultura , Galantamina/isolamento & purificação , Imersão , Liliaceae/crescimento & desenvolvimento , Brotos de Planta/crescimento & desenvolvimento , TemperaturaRESUMO
The common effect of NH4+, NO3-, KH2PO4 and sucrose on the biosynthesis of galanthamine by a Leucojum aestivum shoot culture was studied. Polynominal regression models were elaborated for the description of the galanthamine biosynthesis as a consequence of variation of the investigated variables (NH4+ between 0.20 and 0.54 g/L; NO3- between 1.44 and 3.44 g/L; KH2PO4 between 0.10 and 0.24 g/L, and sucrose between 30.00 and 60.00 g/L). Optimization procedures allowed us to establish the optimal concentrations of the investigated variables and to propose the modified MS nutrient medium, with 4.50 g/L KNO3, 0.89 g/L NH4NO3, 1.25 g/L (NH4)2SO4, 0.10 g/L KH2PO4 and 60 g/L sucrose, for the galanthamine production by a Leucojum aestivum shoot culture. The proposed modified MS medium provided considerable increase of both the production yield and the relative content of the target alkaloid in the alkaloid mixture.
Assuntos
Galantamina/biossíntese , Liliaceae/metabolismo , Brotos de Planta/metabolismo , Alcaloides/biossíntese , Alcaloides/química , Biomassa , Meios de Cultura , Galantamina/química , Galantamina/isolamento & purificação , Cromatografia Gasosa-Espectrometria de Massas , CinéticaRESUMO
The alkaloids of intact plants, calli and shoot-clump cultures of L. aestivum were analyzed by GC-MS. Twenty-four alkaloids were detected. Calli appeared to produce sparse alkaloid profiles in stark contrast to shoot-clumps that had similar profiles to those of the intact plant. Seven shoot-clump strains produced galanthamine predominantly whereas another three were dominated by lycorine. Shoot-clump strains cultivated under light accumulated about two-times more galanthamine (an average of 74 microg/g of dry weight) than those cultivated in darkness (an average of 39 microg/g of dry weight). In comparison to intact plants, the shoot-clumps accumulated 5-times less galanthamine. The high variability of both the galanthamine content (67% and 75% of coefficient of variation under light and darkness conditions, respectively) and alkaloid patterns indicates that the shoot-clump cultures initiated from callus could be used as a tool for improvement of the in vitro cultures.
Assuntos
Alcaloides/análise , Alcaloides/biossíntese , Liliaceae/química , Alcaloides de Amaryllidaceae/metabolismo , Técnicas de Cultura , Galantamina/metabolismo , Cromatografia Gasosa-Espectrometria de Massas , Estrutura Molecular , Fenantridinas/metabolismoRESUMO
The growth and intracellular protein content of lavender (Lavandula vera MM) cell suspension culture was followed along with some antioxidant defense system members-non-enzymatic (rosmarinic acid) and enzymatic [superoxide dismutase (EC 1.15.1.1) and catalase (EC 1.11.1.6)]. It was found that the media content and the cultivation mode strongly influenced the production of plant defense compounds as well as the ratio between non-enzymatic and enzymatic ones. The bioreactor culture contains about two times more rosmarinic acid, superoxide dismutase, and catalase compared to the shake-flask cultivation. These findings are discussed with respect to the relative stress levels and plant antioxidant orchestra system. It was concluded that investigated defense system components (enzymatic and non-enzymatic) were closely associated in a complex balance. The three isoenzyme forms of SOD (Cu/ZnSOD, FeSOD, and MnSOD) in the cells of Lavandula vera were revealed by polyacrylamide gel electrophoresis analysis, and the FeSOD isoform exhibited highest activity.
Assuntos
Antioxidantes/metabolismo , Catalase/metabolismo , Técnicas de Cultura de Células/métodos , Cinamatos/metabolismo , Depsídeos/metabolismo , Lavandula/metabolismo , Superóxido Dismutase/metabolismo , Células Cultivadas , Ácido RosmarínicoRESUMO
This study was conducted to evaluate the radical scavenging capacities of extracts and preparations from a Lavandula vera MM plant cell culture with different rosmarinic acid content and to compare them with pure rosmarinic and caffeic acids as well. The methods, which were used are superoxide anion and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt radicals scavenging assays. Results showed that extracts and preparations from Lavandula vera MM possess strong radical scavengers, as the best both radical scavengers appeared to be the fractions with enriched rosmarinic acid content, obtained after ethylacetate fractioning (47.7% inhibition of superoxide radicals and 14.2 microM 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid equivalents, respectively). These data reveal the possibilities for application of these preparations as antioxidants.
Assuntos
Cinamatos/isolamento & purificação , Cinamatos/farmacologia , Lavandula/química , Fenóis/química , Antioxidantes/química , Antioxidantes/isolamento & purificação , Antioxidantes/farmacologia , Técnicas de Cultura de Células , Depsídeos , Sequestradores de Radicais Livres/farmacologia , Lavandula/citologia , Fenóis/isolamento & purificação , Fenóis/farmacologia , Extratos Vegetais/química , Extratos Vegetais/isolamento & purificação , Superóxidos , Ácido RosmarínicoRESUMO
The influence of elicitation on rosmarinic acid biosynthesis by Lavandula vera MM cell suspension culture was investigated using vanadyl sulfate as an abiotic elicitor. It was established that 12 h after treatment with 25 mg/l vanadyl sulfate the rosmarinic acid production was increased up to 3.92 g/l (2.8 times higher compared to the control cultivation). No significant amounts of rosmarinic acid were detected in the culture medium in comparison with its intracellular content. However, it was observed that the extracellular content of rosmarinic acid is 3.3 times higher compared to the control variant (4 h after treatment at elicitor concentration 25 mg/l).
Assuntos
Cinamatos/metabolismo , Lavandula/metabolismo , Compostos de Vanádio/farmacologia , Divisão Celular/efeitos dos fármacos , Células Cultivadas , Depsídeos , Cinética , Lavandula/citologia , Lavandula/efeitos dos fármacos , Ácido RosmarínicoRESUMO
Stability and radical scavenging activity of betalains, biosynthesized by Beta vulgaris hairy root culture were investigated at different pH values, bile salts concentration, as well as at the in vitro conditions of gastrointestinal tract. It was established that pH below 3 and concentrations of the bile salts up to 4% had no great influence on betalains stability. At the in vitro conditions of gastrointestinal tract betalains are relatively stable, as their radical scavenging activity decrease from 75% inhibition of 2,2-diphenyl-1-picrylhydrazyl (DPPH*) to about 38%.
Assuntos
Beta vulgaris/química , Sequestradores de Radicais Livres/metabolismo , Conteúdo Gastrointestinal/química , Compostos de Amônio Quaternário/metabolismo , Betalaínas , Células Cultivadas , Humanos , Concentração de Íons de Hidrogênio , Modelos Biológicos , Oxirredução , Raízes de Plantas/químicaRESUMO
Studies were conducted on low molecular metabolites (volatiles and polar compounds) produced by Rosa damascena Mill 1803 cell suspension culture, cultivated under different regimes: as a free suspension (in flasks and in bioreactor) and in a two-phase system (in the presence of Amberlite XAD-4 as a second phase). It was established that the main groups of volatiles were hydrocarbons and free acids and their esters and only traces of terpenoids were found. The main components of polar fraction were free acids, especially amino acids and oxidized acids. Depending on the culture conditions, significant differences were established in the amounts of all compounds under study in biomasses, culture media and adsorbed on the second phase (Amberlite XAD-4).
Assuntos
Técnicas de Cultura de Células/métodos , Óleos Voláteis/metabolismo , Óleos de Plantas/metabolismo , Rosa/classificação , Rosa/metabolismo , Células Cultivadas , Caules de Planta/metabolismo , VolatilizaçãoRESUMO
Underivatised alkaloid mixtures extracted from intact plants and in vitro cultures of Leucojum aestivum (Amaryllidaceae) were investigated by capillary GC-MS. Excellent peak resolution for the alkaloids was exhibited and isomers of galanthamine and N-formylnorgalanthamine were well separated. Fourteen alkaloids of galanthamine, lycorine and crinane types were identified, 11 in the intact plants and eight in the in vitro cultures.
Assuntos
Alcaloides/análise , Liliaceae/química , Alcaloides/química , Cromatografia Gasosa-Espectrometria de Massas , Modelos Químicos , Estrutura MolecularRESUMO
The all-round effect of dissolved oxygen concentration, agitation speed, and temperature on the rosmarinic acid production by Lavandula vera MM cell suspension was studied in a 3-L laboratory bioreactor by means of the modified Simplex method. Polynomial regression models were elaborated for description of the process of rosmarinic acid production (Y) in the bioreactor as a consequence of the variation of the dissolved oxygen (X(1)) concentration between 10% and 50%; agitation (X(2)) between 100 and 400 rpm; and temperature (X(3)) between 22 and 30 degrees C. The optimization made it possible to establish the optimal conditions for the biosynthesis of rosmarinic acid by L. vera MM: dissolved oxygen (X(1)*), 50% of air saturation; agitation (X(2)*), 400 rpm; and temperature (X(3)*), 29.9 degrees C, where maximal yield (Y(max)) of 3489.4 mg/L of rosmarinic acid was achieved (2 times higher compared with the shake-flasks cultivation).
Assuntos
Reatores Biológicos , Cinamatos/metabolismo , Lavandula/metabolismo , Depsídeos , Lavandula/citologia , Ácido RosmarínicoRESUMO
Lavandula vera MM cell suspension, grown at 28 degrees C in a 3-l bioreactor, produced rosmarinic acid maximally at 3 g l(-1)) though most biomass (33.2 g dry wt l(-1)) was at 30 degrees C.
Assuntos
Reatores Biológicos/microbiologia , Técnicas de Cultura de Células/métodos , Cinamatos/metabolismo , Lavandula/crescimento & desenvolvimento , Lavandula/metabolismo , Temperatura , Proliferação de Células , Depsídeos , Ácido RosmarínicoRESUMO
Betalains biosynthesis and antiradical scavenging activity were investigated during cultivation of four hairy root cultures of Beta vulgaris, obtained from different cultivars (Bordo, Egyptian, Detroit 2 and Detroit Dark Red). The best producer of betalains was a hairy root culture from Beta vulgaris cv. Detroit Dark Red (13.27 mg/g dry weight total pigment production). The ethanol extract, derived from roots of the same culture grown for 15 days under submerged conditions, showed a high antiradical activity (83% of inhibition of the stable DPPH.).