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Dominique Laurain-mattar - One of the best experts on this subject based on the ideXlab platform.

  • Molecular Identification of Endophytic Bacteria in Leucojum aestivum In Vitro Culture, NMR-Based Metabolomics Study and LC-MS Analysis Leading to Potential Amaryllidaceae Alkaloid Production.
    International journal of molecular sciences, 2021
    Co-Authors: Rosella Spina, Sahar Saliba, François Dupire, Agata Ptak, Alain Hehn, Séverine Piutti, Sophie Poinsignon, Sébastien Leclerc, Sabine Bouguet-bonnet, Dominique Laurain-mattar
    Abstract:

    In this study, endophytic bacteria belonging to the Bacillus genus were isolated from in vitro bulblets of Leucojum aestivum and their ability to produce Amaryllidaceae Alkaloids was studied. Proton Nuclear Magnetic Resonance (1H NMR)-based metabolomics combined with multivariate data analysis was chosen to compare the metabolism of this plant (in vivo bulbs, in vitro bulblets) with those of the endophytic bacteria community. Primary metabolites were quantified by quantitative 1H NMR (qNMR) method. The results showed that tyrosine, one precursor of the Amaryllidaceae Alkaloid biosynthesis pathway, was higher in endophytic extract compared to plant extract. In total, 22 compounds were identified including five molecules common to plant and endophyte extracts (tyrosine, isoleucine, valine, fatty acids and tyramine). In addition, endophytic extracts were analyzed using Liquid Chromatography-Mass Spectrometry (LC-MS) and Gas Chromatography-Mass Spectrometry (GC-MS) for the identification of compounds in very low concentrations. Five Amaryllidaceae Alkaloids were detected in the extracts of endophytic bacteria. Lycorine, previously detected by 1H NMR, was confirmed with LC-MS analysis. Tazettine, pseudolycorine, acetylpseudolycorine, 1,2-dihydro-chlidanthine were also identified by LC-MS using the positive ionization mode or by GC-MS. In addition, 11 primary metabolites were identified in the endophytic extracts such as tyramine, which was obtained by decarboxylation of tyrosine. Thus, Bacillus sp. isolated from L. aestivum bulblets synthesized some primary and specialized metabolites in common with the L.aestivum plant. These endophytic bacteria are an interesting new approach for producing the Amaryllidaceae Alkaloid such as lycorine.

  • Carbohydrates stimulated Amaryllidaceae Alkaloids biosynthesis in Leucojum aestivum L. plants cultured in RITA® bioreactor.
    PeerJ, 2020
    Co-Authors: Agata Ptak, Dominique Laurain-mattar, Rosella Spina, Emilia Morańska, Edyta Skrzypek, Marzena Warchoł, Magdalena Simlat
    Abstract:

    Background Leucojum aestivum L. is an important medicinal plant which produces Amaryllidaceae Alkaloids, especially galanthamine and lycorine. Research is currently exploring the possibility of producing these Alkaloids using biotechnological methods, including in vitro cultures. The biosynthesis of Alkaloids may be affected by the types and concentrations of carbohydrate sources used in the medium. In the present investigation we performed such studies on in vitro cultures of L. aestivum with a view to obtaining plant material of good quality, characterized, in particular, by a high content of valuable Amaryllidaceae Alkaloids. Methods We examined the effects of various types of carbohydrate sources-sucrose, glucose, fructose and maltose-at different concentrations (30, 60 and 90 g/L)-on the quality of L. aestivum plants grown in the RITA® bioreactor. The plants' quality was assessed by their biomass increments, as well by as analysing photosynthetic pigments, endogenous sugar, phenolics and Amaryllidaceae Alkaloid content. We also investigated the effect of sugars on the activity of the antioxidant enzymes catalase (CAT), peroxidase (POD) and superoxide dismutase (SOD). Results The highest biomass increments were observed in plants cultivated in the medium containing 90 g/L sucrose. The highest CAT activity was noted in cultures growing in the medium supplemented with 90 g/L maltose, while the highest POD activity was observed in the presence of 90 g/L fructose and 60 g/L maltose. No differences in SOD activity were observed. Moreover, the sugars did not affect the contents of chlorophyll a and carotenoids, whereas the highest amount of chlorophyll b was recorded in plants growing in the medium with 60 g/L maltose. No statistically significant differences were observed in the contents of endogenous sugars and phenolics in any in vitro conditions. However, the addition of sugar had a decisive effect on the biosynthesis of the Amaryllidaceae Alkaloids. The highest distribution of Alkaloids occurred in plants cultured in the medium containing 60 g/L sucrose. Six Amaryllidaceae Alkaloids were detected in the plant tissue. The addition of 30 g/L fructose in the medium resulted in the accumulation of five Alkaloids, including ismine, which was not identified in other analysed tissues. The highest concentration of galanthamine was observed in plants cultured in the presence of 30 g/L fructose and 60 g/L sucrose (39.2 and 37.5 µg/g of dry weight (DW), respectively). The plants grown in the medium containing 60 g/L sucrose exhibited the highest lycorine content (1048 µg/g of DW). Conclusions The type and concentration of sugar used in the medium have an essential influence on the biosynthesis of Amaryllidaceae Alkaloids in L. aestivum plants cultured in a RITA® bioreactor. The results point to an interesting approach for commercial production of galanthamine and lycorine.

  • Carbohydrates stimulated Amaryllidaceae Alkaloids biosynthesis in Leucojum aestivum L. plants cultured in RITA ® bioreactor
    PeerJ, 2020
    Co-Authors: Agata Ptak, Dominique Laurain-mattar, Rosella Spina, Emilia Morańska, Edyta Skrzypek, Marzena Warchoł, Magdalena Simlat
    Abstract:

    Background: Leucojum aestivum L. is an important medicinal plant which produces Amaryllidaceae Alkaloids, especially galanthamine and lycorine. Research is currently exploring the possibility of producing these Alkaloids using biotechnological methods, including in vitro cultures. The biosynthesis of Alkaloids may be affected by the types and concentrations of carbohydrate sources used in the medium. In the present investigation we performed such studies on in vitro cultures of L. aestivum with a view to obtaining plant material of good quality, characterized, in particular, by a high content of valuable Amaryllidaceae Alkaloids. Methods: We examined the effects of various types of carbohydrate sources-sucrose, glucose, fructose and maltose-at different concentrations (30, 60 and 90 g/L)-on the quality of L. aestivum plants grown in the RITA Ò bioreactor. The plants' quality was assessed by their biomass increments, as well by as analysing photosynthetic pigments, endogenous sugar, phenolics and Amaryllidaceae Alkaloid content. We also investigated the effect of sugars on the activity of the antioxidant enzymes catalase (CAT), peroxidase (POD) and superoxide dismutase (SOD). Results: The highest biomass increments were observed in plants cultivated in the medium containing 90 g/L sucrose. The highest CAT activity was noted in cultures growing in the medium supplemented with 90 g/L maltose, while the highest POD activity was observed in the presence of 90 g/L fructose and 60 g/L maltose. No differences in SOD activity were observed. Moreover, the sugars did not affect the contents of chlorophyll a and carotenoids, whereas the highest amount of chlorophyll b was recorded in plants growing in the medium with 60 g/L maltose. No statistically significant differences were observed in the contents of endogenous sugars and phenolics in any in vitro conditions. However, the addition of sugar had a decisive effect on the biosynthesis of the Amaryllidaceae Alkaloids. The highest distribution of Alkaloids occurred in plants cultured in the medium containing 60 g/L sucrose. Six Amaryllidaceae Alkaloids were detected in the plant tissue. The addition of 30 g/L fructose in the medium resulted in the accumulation of five Alkaloids, including ismine, which was not identified in other analysed tissues. The highest

  • Exogenous melatonin stimulated Amaryllidaceae Alkaloid biosynthesis in in vitro cultures of Leucojum aestivum L
    Industrial Crops and Products, 2019
    Co-Authors: Agata Ptak, Magdalena Simlat, Emilia Morańska, Edyta Skrzypek, Marzena Warchoł, Ameneh Tarakemeh, Dominique Laurain-mattar
    Abstract:

    Abstract Leucojum aestivum L. is a highly demanding pharmaceutical plant which is a natural source of pharmacologically active Amaryllidaceae Alkaloids, especially galanthamine and lycorine. To establish an efficient method for the production of these Alkaloids Leucojum aestivum plants derived from somatic embryos were subjected to various concentrations of melatonin (N-acetyl-5-methoxy tryptamine), NaCl and melatonin combined with NaCl. The maximum L. aestivum plant biomass was obtained when 10 μM of melatonin was applied to the medium. These plants were characterised by normal growth and did not reveal signs of browning. Enrichment of the medium with NaCl caused a reduction in plant biomass, plant pigment contents (chlorophyll a, chlorophyll b and carotenoids) and an increase in the activities of antioxidant enzymes catalase (CAT), peroxidase (POD) and superoxide dismutase (SOD). The addition of melatonin to the medium containing NaCl neutralised the negative effect of salt stress. Moreover, the stimulating effect of melatonin on Amaryllidaceae Alkaloids biosynthesis was observed. In plants grown in the medium enriched with melatonin the galanthamine content was, on average, 58.6 times higher, and the lycorine content was 1.5 times higher as compared with the control. In vitro plants obtained from the medium supplemented with melatonin also showed the highest distribution of Alkaloids. It is noteworthy that only the plants grown in the medium with melatonin showed the presence of tazettine and chlidanthine, 1,2-dihydro-. This study presents a promising biotechnological approach for the commercial production of Alkaloids, including galanthamine and lycorine.

  • Amaryllidaceae Alkaloid Accumulation by Plant In Vitro Systems
    Bioprocessing of Plant In Vitro Systems, 2017
    Co-Authors: Dominique Laurain-mattar, Aleksandra Ptak
    Abstract:

    Over 300 Alkaloids possessing a wide range of biological activities have been isolated from plants belonging to Amaryllidaceae family. Galanthamine, used for the palliative treatment of Alzheimer’s disease, is the only one commercialized. The biodisponibility of Amaryllidaceae Alkaloids is low. In vitro culture offers an alternative interesting approach for the biotechnological production of these valuable Alkaloids. The feeding with different exogenous factors of Leucojum aestivum, L. aestivum ‘Gravety Giant,’ and Narcissus in vitro cultures was reported, and the effects on the biosynthetic pathway of both galanthamine and lycorine were studied.

Agata Ptak - One of the best experts on this subject based on the ideXlab platform.

  • Molecular Identification of Endophytic Bacteria in Leucojum aestivum In Vitro Culture, NMR-Based Metabolomics Study and LC-MS Analysis Leading to Potential Amaryllidaceae Alkaloid Production.
    International journal of molecular sciences, 2021
    Co-Authors: Rosella Spina, Sahar Saliba, François Dupire, Agata Ptak, Alain Hehn, Séverine Piutti, Sophie Poinsignon, Sébastien Leclerc, Sabine Bouguet-bonnet, Dominique Laurain-mattar
    Abstract:

    In this study, endophytic bacteria belonging to the Bacillus genus were isolated from in vitro bulblets of Leucojum aestivum and their ability to produce Amaryllidaceae Alkaloids was studied. Proton Nuclear Magnetic Resonance (1H NMR)-based metabolomics combined with multivariate data analysis was chosen to compare the metabolism of this plant (in vivo bulbs, in vitro bulblets) with those of the endophytic bacteria community. Primary metabolites were quantified by quantitative 1H NMR (qNMR) method. The results showed that tyrosine, one precursor of the Amaryllidaceae Alkaloid biosynthesis pathway, was higher in endophytic extract compared to plant extract. In total, 22 compounds were identified including five molecules common to plant and endophyte extracts (tyrosine, isoleucine, valine, fatty acids and tyramine). In addition, endophytic extracts were analyzed using Liquid Chromatography-Mass Spectrometry (LC-MS) and Gas Chromatography-Mass Spectrometry (GC-MS) for the identification of compounds in very low concentrations. Five Amaryllidaceae Alkaloids were detected in the extracts of endophytic bacteria. Lycorine, previously detected by 1H NMR, was confirmed with LC-MS analysis. Tazettine, pseudolycorine, acetylpseudolycorine, 1,2-dihydro-chlidanthine were also identified by LC-MS using the positive ionization mode or by GC-MS. In addition, 11 primary metabolites were identified in the endophytic extracts such as tyramine, which was obtained by decarboxylation of tyrosine. Thus, Bacillus sp. isolated from L. aestivum bulblets synthesized some primary and specialized metabolites in common with the L.aestivum plant. These endophytic bacteria are an interesting new approach for producing the Amaryllidaceae Alkaloid such as lycorine.

  • Carbohydrates stimulated Amaryllidaceae Alkaloids biosynthesis in Leucojum aestivum L. plants cultured in RITA® bioreactor.
    PeerJ, 2020
    Co-Authors: Agata Ptak, Dominique Laurain-mattar, Rosella Spina, Emilia Morańska, Edyta Skrzypek, Marzena Warchoł, Magdalena Simlat
    Abstract:

    Background Leucojum aestivum L. is an important medicinal plant which produces Amaryllidaceae Alkaloids, especially galanthamine and lycorine. Research is currently exploring the possibility of producing these Alkaloids using biotechnological methods, including in vitro cultures. The biosynthesis of Alkaloids may be affected by the types and concentrations of carbohydrate sources used in the medium. In the present investigation we performed such studies on in vitro cultures of L. aestivum with a view to obtaining plant material of good quality, characterized, in particular, by a high content of valuable Amaryllidaceae Alkaloids. Methods We examined the effects of various types of carbohydrate sources-sucrose, glucose, fructose and maltose-at different concentrations (30, 60 and 90 g/L)-on the quality of L. aestivum plants grown in the RITA® bioreactor. The plants' quality was assessed by their biomass increments, as well by as analysing photosynthetic pigments, endogenous sugar, phenolics and Amaryllidaceae Alkaloid content. We also investigated the effect of sugars on the activity of the antioxidant enzymes catalase (CAT), peroxidase (POD) and superoxide dismutase (SOD). Results The highest biomass increments were observed in plants cultivated in the medium containing 90 g/L sucrose. The highest CAT activity was noted in cultures growing in the medium supplemented with 90 g/L maltose, while the highest POD activity was observed in the presence of 90 g/L fructose and 60 g/L maltose. No differences in SOD activity were observed. Moreover, the sugars did not affect the contents of chlorophyll a and carotenoids, whereas the highest amount of chlorophyll b was recorded in plants growing in the medium with 60 g/L maltose. No statistically significant differences were observed in the contents of endogenous sugars and phenolics in any in vitro conditions. However, the addition of sugar had a decisive effect on the biosynthesis of the Amaryllidaceae Alkaloids. The highest distribution of Alkaloids occurred in plants cultured in the medium containing 60 g/L sucrose. Six Amaryllidaceae Alkaloids were detected in the plant tissue. The addition of 30 g/L fructose in the medium resulted in the accumulation of five Alkaloids, including ismine, which was not identified in other analysed tissues. The highest concentration of galanthamine was observed in plants cultured in the presence of 30 g/L fructose and 60 g/L sucrose (39.2 and 37.5 µg/g of dry weight (DW), respectively). The plants grown in the medium containing 60 g/L sucrose exhibited the highest lycorine content (1048 µg/g of DW). Conclusions The type and concentration of sugar used in the medium have an essential influence on the biosynthesis of Amaryllidaceae Alkaloids in L. aestivum plants cultured in a RITA® bioreactor. The results point to an interesting approach for commercial production of galanthamine and lycorine.

  • Carbohydrates stimulated Amaryllidaceae Alkaloids biosynthesis in Leucojum aestivum L. plants cultured in RITA ® bioreactor
    PeerJ, 2020
    Co-Authors: Agata Ptak, Dominique Laurain-mattar, Rosella Spina, Emilia Morańska, Edyta Skrzypek, Marzena Warchoł, Magdalena Simlat
    Abstract:

    Background: Leucojum aestivum L. is an important medicinal plant which produces Amaryllidaceae Alkaloids, especially galanthamine and lycorine. Research is currently exploring the possibility of producing these Alkaloids using biotechnological methods, including in vitro cultures. The biosynthesis of Alkaloids may be affected by the types and concentrations of carbohydrate sources used in the medium. In the present investigation we performed such studies on in vitro cultures of L. aestivum with a view to obtaining plant material of good quality, characterized, in particular, by a high content of valuable Amaryllidaceae Alkaloids. Methods: We examined the effects of various types of carbohydrate sources-sucrose, glucose, fructose and maltose-at different concentrations (30, 60 and 90 g/L)-on the quality of L. aestivum plants grown in the RITA Ò bioreactor. The plants' quality was assessed by their biomass increments, as well by as analysing photosynthetic pigments, endogenous sugar, phenolics and Amaryllidaceae Alkaloid content. We also investigated the effect of sugars on the activity of the antioxidant enzymes catalase (CAT), peroxidase (POD) and superoxide dismutase (SOD). Results: The highest biomass increments were observed in plants cultivated in the medium containing 90 g/L sucrose. The highest CAT activity was noted in cultures growing in the medium supplemented with 90 g/L maltose, while the highest POD activity was observed in the presence of 90 g/L fructose and 60 g/L maltose. No differences in SOD activity were observed. Moreover, the sugars did not affect the contents of chlorophyll a and carotenoids, whereas the highest amount of chlorophyll b was recorded in plants growing in the medium with 60 g/L maltose. No statistically significant differences were observed in the contents of endogenous sugars and phenolics in any in vitro conditions. However, the addition of sugar had a decisive effect on the biosynthesis of the Amaryllidaceae Alkaloids. The highest distribution of Alkaloids occurred in plants cultured in the medium containing 60 g/L sucrose. Six Amaryllidaceae Alkaloids were detected in the plant tissue. The addition of 30 g/L fructose in the medium resulted in the accumulation of five Alkaloids, including ismine, which was not identified in other analysed tissues. The highest

  • Exogenous melatonin stimulated Amaryllidaceae Alkaloid biosynthesis in in vitro cultures of Leucojum aestivum L
    Industrial Crops and Products, 2019
    Co-Authors: Agata Ptak, Magdalena Simlat, Emilia Morańska, Edyta Skrzypek, Marzena Warchoł, Ameneh Tarakemeh, Dominique Laurain-mattar
    Abstract:

    Abstract Leucojum aestivum L. is a highly demanding pharmaceutical plant which is a natural source of pharmacologically active Amaryllidaceae Alkaloids, especially galanthamine and lycorine. To establish an efficient method for the production of these Alkaloids Leucojum aestivum plants derived from somatic embryos were subjected to various concentrations of melatonin (N-acetyl-5-methoxy tryptamine), NaCl and melatonin combined with NaCl. The maximum L. aestivum plant biomass was obtained when 10 μM of melatonin was applied to the medium. These plants were characterised by normal growth and did not reveal signs of browning. Enrichment of the medium with NaCl caused a reduction in plant biomass, plant pigment contents (chlorophyll a, chlorophyll b and carotenoids) and an increase in the activities of antioxidant enzymes catalase (CAT), peroxidase (POD) and superoxide dismutase (SOD). The addition of melatonin to the medium containing NaCl neutralised the negative effect of salt stress. Moreover, the stimulating effect of melatonin on Amaryllidaceae Alkaloids biosynthesis was observed. In plants grown in the medium enriched with melatonin the galanthamine content was, on average, 58.6 times higher, and the lycorine content was 1.5 times higher as compared with the control. In vitro plants obtained from the medium supplemented with melatonin also showed the highest distribution of Alkaloids. It is noteworthy that only the plants grown in the medium with melatonin showed the presence of tazettine and chlidanthine, 1,2-dihydro-. This study presents a promising biotechnological approach for the commercial production of Alkaloids, including galanthamine and lycorine.

  • kinetic study of the rearrangement of deuterium labeled 4 o methylnorbelladine in leucojum aestivum shoot cultures by mass spectrometry influence of precursor feeding on Amaryllidaceae Alkaloid accumulation
    Journal of Natural Products, 2011
    Co-Authors: Anna El Tahchy, François Dupire, Agata Ptak, Michel Boisbrun, Françoise Chrétien, Max Henry, Yves Chapleur, Elvina Barre, Catherine Guillou, Dominique Laurainmattar
    Abstract:

    Alkaloids from plants of the family Amaryllidaceae have important pharmacological properties and can be regarded as derivatives of the common precursor 4'-O-methylnorbelladine (6) via intramolecular oxidative phenol coupling. Their biosynthetic pathway, particularly in Leucojum aestivum, has not yet been totally elucidated. Therefore, shoot cultures of this plant were subcultured in medium containing the labeled precursor 4'-O-methyl-d(3)-norbelladine (3) at various concentrations (0.05, 0.10, and 0.20 g/L) and were incubated for various periods of time (15, 30, and 40 days). The aim of this work was to study the influence of this precursor on both labeled and native Alkaloid accumulation. Biotransformation into galanthamine (1) and lycorine (2) in shoot cultures was demonstrated using HPLC coupled to mass spectrometry. A maximal amount of 0.16% of 1 referred to the dry weight was obtained at day 15 in shoots fed with 0.10 g/L of precursor. In addition, a 20.5% dry weight of 2 was reached after 40 days of feeding with 0.20 g/L of precursor.

Magdalena Simlat - One of the best experts on this subject based on the ideXlab platform.

  • Carbohydrates stimulated Amaryllidaceae Alkaloids biosynthesis in Leucojum aestivum L. plants cultured in RITA® bioreactor.
    PeerJ, 2020
    Co-Authors: Agata Ptak, Dominique Laurain-mattar, Rosella Spina, Emilia Morańska, Edyta Skrzypek, Marzena Warchoł, Magdalena Simlat
    Abstract:

    Background Leucojum aestivum L. is an important medicinal plant which produces Amaryllidaceae Alkaloids, especially galanthamine and lycorine. Research is currently exploring the possibility of producing these Alkaloids using biotechnological methods, including in vitro cultures. The biosynthesis of Alkaloids may be affected by the types and concentrations of carbohydrate sources used in the medium. In the present investigation we performed such studies on in vitro cultures of L. aestivum with a view to obtaining plant material of good quality, characterized, in particular, by a high content of valuable Amaryllidaceae Alkaloids. Methods We examined the effects of various types of carbohydrate sources-sucrose, glucose, fructose and maltose-at different concentrations (30, 60 and 90 g/L)-on the quality of L. aestivum plants grown in the RITA® bioreactor. The plants' quality was assessed by their biomass increments, as well by as analysing photosynthetic pigments, endogenous sugar, phenolics and Amaryllidaceae Alkaloid content. We also investigated the effect of sugars on the activity of the antioxidant enzymes catalase (CAT), peroxidase (POD) and superoxide dismutase (SOD). Results The highest biomass increments were observed in plants cultivated in the medium containing 90 g/L sucrose. The highest CAT activity was noted in cultures growing in the medium supplemented with 90 g/L maltose, while the highest POD activity was observed in the presence of 90 g/L fructose and 60 g/L maltose. No differences in SOD activity were observed. Moreover, the sugars did not affect the contents of chlorophyll a and carotenoids, whereas the highest amount of chlorophyll b was recorded in plants growing in the medium with 60 g/L maltose. No statistically significant differences were observed in the contents of endogenous sugars and phenolics in any in vitro conditions. However, the addition of sugar had a decisive effect on the biosynthesis of the Amaryllidaceae Alkaloids. The highest distribution of Alkaloids occurred in plants cultured in the medium containing 60 g/L sucrose. Six Amaryllidaceae Alkaloids were detected in the plant tissue. The addition of 30 g/L fructose in the medium resulted in the accumulation of five Alkaloids, including ismine, which was not identified in other analysed tissues. The highest concentration of galanthamine was observed in plants cultured in the presence of 30 g/L fructose and 60 g/L sucrose (39.2 and 37.5 µg/g of dry weight (DW), respectively). The plants grown in the medium containing 60 g/L sucrose exhibited the highest lycorine content (1048 µg/g of DW). Conclusions The type and concentration of sugar used in the medium have an essential influence on the biosynthesis of Amaryllidaceae Alkaloids in L. aestivum plants cultured in a RITA® bioreactor. The results point to an interesting approach for commercial production of galanthamine and lycorine.

  • Carbohydrates stimulated Amaryllidaceae Alkaloids biosynthesis in Leucojum aestivum L. plants cultured in RITA ® bioreactor
    PeerJ, 2020
    Co-Authors: Agata Ptak, Dominique Laurain-mattar, Rosella Spina, Emilia Morańska, Edyta Skrzypek, Marzena Warchoł, Magdalena Simlat
    Abstract:

    Background: Leucojum aestivum L. is an important medicinal plant which produces Amaryllidaceae Alkaloids, especially galanthamine and lycorine. Research is currently exploring the possibility of producing these Alkaloids using biotechnological methods, including in vitro cultures. The biosynthesis of Alkaloids may be affected by the types and concentrations of carbohydrate sources used in the medium. In the present investigation we performed such studies on in vitro cultures of L. aestivum with a view to obtaining plant material of good quality, characterized, in particular, by a high content of valuable Amaryllidaceae Alkaloids. Methods: We examined the effects of various types of carbohydrate sources-sucrose, glucose, fructose and maltose-at different concentrations (30, 60 and 90 g/L)-on the quality of L. aestivum plants grown in the RITA Ò bioreactor. The plants' quality was assessed by their biomass increments, as well by as analysing photosynthetic pigments, endogenous sugar, phenolics and Amaryllidaceae Alkaloid content. We also investigated the effect of sugars on the activity of the antioxidant enzymes catalase (CAT), peroxidase (POD) and superoxide dismutase (SOD). Results: The highest biomass increments were observed in plants cultivated in the medium containing 90 g/L sucrose. The highest CAT activity was noted in cultures growing in the medium supplemented with 90 g/L maltose, while the highest POD activity was observed in the presence of 90 g/L fructose and 60 g/L maltose. No differences in SOD activity were observed. Moreover, the sugars did not affect the contents of chlorophyll a and carotenoids, whereas the highest amount of chlorophyll b was recorded in plants growing in the medium with 60 g/L maltose. No statistically significant differences were observed in the contents of endogenous sugars and phenolics in any in vitro conditions. However, the addition of sugar had a decisive effect on the biosynthesis of the Amaryllidaceae Alkaloids. The highest distribution of Alkaloids occurred in plants cultured in the medium containing 60 g/L sucrose. Six Amaryllidaceae Alkaloids were detected in the plant tissue. The addition of 30 g/L fructose in the medium resulted in the accumulation of five Alkaloids, including ismine, which was not identified in other analysed tissues. The highest

  • Exogenous melatonin stimulated Amaryllidaceae Alkaloid biosynthesis in in vitro cultures of Leucojum aestivum L
    Industrial Crops and Products, 2019
    Co-Authors: Agata Ptak, Magdalena Simlat, Emilia Morańska, Edyta Skrzypek, Marzena Warchoł, Ameneh Tarakemeh, Dominique Laurain-mattar
    Abstract:

    Abstract Leucojum aestivum L. is a highly demanding pharmaceutical plant which is a natural source of pharmacologically active Amaryllidaceae Alkaloids, especially galanthamine and lycorine. To establish an efficient method for the production of these Alkaloids Leucojum aestivum plants derived from somatic embryos were subjected to various concentrations of melatonin (N-acetyl-5-methoxy tryptamine), NaCl and melatonin combined with NaCl. The maximum L. aestivum plant biomass was obtained when 10 μM of melatonin was applied to the medium. These plants were characterised by normal growth and did not reveal signs of browning. Enrichment of the medium with NaCl caused a reduction in plant biomass, plant pigment contents (chlorophyll a, chlorophyll b and carotenoids) and an increase in the activities of antioxidant enzymes catalase (CAT), peroxidase (POD) and superoxide dismutase (SOD). The addition of melatonin to the medium containing NaCl neutralised the negative effect of salt stress. Moreover, the stimulating effect of melatonin on Amaryllidaceae Alkaloids biosynthesis was observed. In plants grown in the medium enriched with melatonin the galanthamine content was, on average, 58.6 times higher, and the lycorine content was 1.5 times higher as compared with the control. In vitro plants obtained from the medium supplemented with melatonin also showed the highest distribution of Alkaloids. It is noteworthy that only the plants grown in the medium with melatonin showed the presence of tazettine and chlidanthine, 1,2-dihydro-. This study presents a promising biotechnological approach for the commercial production of Alkaloids, including galanthamine and lycorine.

Wolfang Oppolzer - One of the best experts on this subject based on the ideXlab platform.

Toni M. Kutchan - One of the best experts on this subject based on the ideXlab platform.

  • Identification of a Noroxomaritidine Reductase with Amaryllidaceae Alkaloid Biosynthesis Related Activities.
    The Journal of biological chemistry, 2016
    Co-Authors: Matthew B. Kilgore, Cynthia K. Holland, Joseph M. Jez, Toni M. Kutchan
    Abstract:

    Abstract Amaryllidaceae Alkaloids are a large group of plant natural products with over 300 documented structures and diverse biological activities. Several groups of Amaryllidaceae Alkaloids including the haemanthamine and crinine type Alkaloids show promise as anticancer agents. Two reduction reactions are required for the production of these compounds: the reduction of norcraugsodine to norbelladine and the reduction of noroxomaritidine to normaritidine, with the enantiomer of noroxomaritidine dictating whether the derivatives will be the crinine type or haemanthamine type. It is also possible for the carbon-carbon double bond of noroxomaritidine to be reduced, forming the precursor for maritinamine or elwesine depending on the enantiomer reduced to an oxomaritinamine product. In this study, a short-chain alcohol dehydrogenase/reductase that co-expresses with the previously discovered norbelladine 4'-O-methyltransferase from Narcissus sp. and Galanthus spp. was cloned and expressed in E. coli. Biochemical analyses and x-ray crystallography indicates that this protein functions as a noroxomaritidine reductase that forms oxomaritinamine from noroxomaritidine through a carbon-carbon double bond reduction. The enzyme also reduces norcraugsodine to norbelladine with a 400-fold lower specific activity. These studies identify a missing step in the biosynthesis of this pharmacologically important class of plant natural products.

  • The Amaryllidaceae Alkaloids: biosynthesis and methods for enzyme discovery
    Phytochemistry Reviews, 2016
    Co-Authors: Matthew B. Kilgore, Toni M. Kutchan
    Abstract:

    Amaryllidaceae Alkaloids are an example of the vast diversity of secondary metabolites with great therapeutic promise. The identification of novel compounds in this group with over 300 known structures continues to be an area of active study. The recent identification of norbelladine 4′- O -methyltransferase ( N4OMT ), an Amaryllidaceae Alkaloid biosynthetic enzyme, and the assembly of transcriptomes for Narcissus sp. aff. pseudonarcissus and Lycoris aurea highlight the potential for discovery of Amaryllidaceae Alkaloid biosynthetic genes with new technologies. Recent technical advances of interest include those in enzymology, next generation sequencing, genetic modification, nuclear magnetic resonance spectroscopy, and mass spectrometry.

  • Identification of a Noroxomaritidine Reductase with Amaryllidaceae Alkaloid Biosynthesis Related Activities.
    The Journal of biological chemistry, 2016
    Co-Authors: Matthew Kilgore, Joseph M. Jez, Cynthia K. Holland, Toni M. Kutchan
    Abstract:

    Amaryllidaceae Alkaloids are a large group of plant natural products with over 300 documented structures and diverse biological activities. Several groups of Amaryllidaceae Alkaloids including the hemanthamine- and crinine-type Alkaloids show promise as anticancer agents. Two reduction reactions are required for the production of these compounds: the reduction of norcraugsodine to norbelladine and the reduction of noroxomaritidine to normaritidine, with the enantiomer of noroxomaritidine dictating whether the derivatives will be the crinine-type or hemanthamine-type. It is also possible for the carbon-carbon double bond of noroxomaritidine to be reduced, forming the precursor for maritinamine or elwesine depending on the enantiomer reduced to an oxomaritinamine product. In this study, a short chain alcohol dehydrogenase/reductase that co-expresses with the previously discovered norbelladine 4′-O-methyltransferase from Narcissus sp. and Galanthus spp. was cloned and expressed in Escherichia coli. Biochemical analyses and x-ray crystallography indicates that this protein functions as a noroxomaritidine reductase that forms oxomaritinamine from noroxomaritidine through a carbon-carbon double bond reduction. The enzyme also reduces norcraugsodine to norbelladine with a 400-fold lower specific activity. These studies identify a missing step in the biosynthesis of this pharmacologically important class of plant natural products.

  • CYP96T1 of Narcissus sp. aff. pseudonarcissus Catalyzes Formation of the Para-Para’ C-C Phenol Couple in the Amaryllidaceae Alkaloids
    Frontiers Media S.A., 2016
    Co-Authors: Matthew Ekilgore, Megan M. Augustin, Gregory D. May, John A. Crow, Toni M. Kutchan
    Abstract:

    The Amaryllidaceae Alkaloids are a family of amino acid derived Alkaloids with many biological activities; examples include haemanthamine, haemanthidine, galanthamine, lycorine, and maritidine. Central to the biosynthesis of the majority of these Alkaloids is a C-C phenol-coupling reaction that can have para-para’, para-ortho’, or ortho-para’ regiospecificity. Through comparative transcriptomics of Narcissus sp. aff. pseudonarcissus, Galanthus sp., and Galanthus elwesii we have identified a para-para’ C-C phenol coupling cytochrome P450, CYP96T1, capable of forming the products (10bR,4aS)-noroxomaritidine and (10bS,4aR)-noroxomaritidine from 4’-O-methylnorbelladine. CYP96T1 was also shown to catalyzed formation of the para-ortho’ phenol coupled product, N-demethylnarwedine, as less than 1 % of the total product. CYP96T1 co-expresses with the previously characterized norbelladine 4’-O-methyltransferase. The discovery of CYP96T1 is of special interest because it catalyzes the first major branch in Amaryllidaceae Alkaloid biosynthesis. CYP96T1 is also the first phenol-coupling enzyme characterized from a monocot