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The aim of this study was to produce Erns protein of bovine viral diarrhea virus (BVDV) by using suspensively cultured CHO cells expression system and to analyze the immunogenicity of the purified Erns protein. In this study, the recombinant eukaryotic expression plasmid pcDNA3.1-BVDV-Erns was constructed based on the gene sequence of BVDV-1 NADL strain. The Erns protein was secreted and expressed in cells supernatant after transfecting the recombinant expression plasmid pcDNA3.1-BVDV-Erns into CHO cells. The expression and purification of the Erns protein was analyzed by SDS-PAGE, the reactivity was determined with anti-His monoclonal antibodies and BVDV positive serum with Western blotting. Immunogenicity analysis of the Erns protein was determined after immunizing New Zealand white rabbits, and the serum antibodies were tested by indirect ELISA (iELISA) and indirect immunofluorescence (IFA). The serum neutralizing titer of the immunized rabbits was determined by virus neutralization test. The concentration of the purified Erns protein was up to 0.886 mg/mL by BCA protein quantification kit. The results showed that the Erns protein could be detected with anti-His monoclonal antibodies and anti-BVDV sera. Serum antibodies could be detected by iELISA on the 7th day post-prime immunization, and the antibody level was maintained at a high titer until the 28th day post-immunization. The antibody titer was 1:128 000. Furthermore, the expression of the Erns protein in BVDV-infected MDBK cells could be detected with immunized rabbits sera by IFA. Moreover, antigen-specific neutralizing antibodies of 2.71 log10 was induced in rabbits. In this study, purified BVDV Erns protein was successfully produced using CHO suspension culture system, and the recombinant protein was proved to have a good immunogenicity, which may facilitate the development of BVD diagnosis method and novel subunit vaccine.
Subject(s)
Rabbits , Animals , Cricetinae , Cricetulus , CHO Cells , Antibodies, Viral , Diarrhea Viruses, Bovine Viral/genetics , Antibodies, Monoclonal/genetics , Diarrhea , Viral Vaccines/geneticsABSTRACT
Centella asiatica is an important medicinal plant which contains various phytocompounds. Asiatic acid and asiaticosideare two major compounds which are responsible for its various pharmaceutical activities. The present study analyzesthe effect of elicitor, i.e., methyl jasmonate on the synthesis of asiaticoside and asiatic acid (ATA) in shoot, callus, andcell suspension cultures of C. asiatica. A high-performance liquid chromatography analysis showed that the elicitationwith 100 µM concentration of methyl jasmonate enhanced asiaticoside content by 69-fold in callus culture, 39-fold inshoot cultures, and ATA by 1.9-fold in cell suspension culture. Thus, elicitation with methyl jasmonate is an effectivemethod of increasing the rate of biosynthesis of asiaticoside and ATA in plant cell cultures of C. asiatica
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Objective: To establish the plant tissue culture system of Cistanche tubulosa, and determine the effect of drought stress on accumulation of two respective phenylethanoid glycosides in it. Methods The major chemical constituents of C. tubulosa by tissue culture were analyzed by HPLC-UV and HR-MS. The cell growth curves were also determined. In addition, the effects of drought stress on the phenylethanoid glycosides (echinacoside and acteoside) content in the tissue culture system of C. tubulosa were also studied by using NaCl, mannitol and PEG6000 as osmotic regulators, respectively. Results:Chemical constituents analyses revealed that callus and suspension cultures of C. tubulosa could produce the respective phenylethanoid glycosides of echinacoside and acteoside as in wild plant; Cell growth curves indicated that 30 d were the optimum culture period of callus culture; The cell growth rate and the accumulation of echinacoside and acteoside were mostly inhibited when the callus cells were under drought stress induced by NaCl or mannitol. Meanwhile, the accumulation of echinacoside and acteoside in cell suspension culture of C. tubulosa could be effectively enhanced by treatment with PEG6000. The maximum biomass of echinacoside and acteoside could reach to (1.07 ± 0.10) g/L and (0.12 ± 0.01) g/L 15 d after induction, respectively. And their contents were 20.94% and 2.27% separately based on the cell dry weight (DW) after 15 d of treatment with 6% PEG6000, which were 1.29 and 1.19 fold higher than the control group. Conclusion:Drought stress induced by PEG6000 could effectively enhance the accumulation of echinacoside and acteoside in cell suspension culture of C. tubulosa.
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Cell suspension cultures of Thevetia peruviana were established under dark for 19 days to investigate kinetic behavior related to biomass, substrate, cardiac glycoside, polyphenols, reactive oxygen species and anti-oxidant activity. The results showed high biomass production (18.80gDW/L) as well as sucrose consumption in 7 days. Preferential glucose over fructose consumption was observed. Intracellular production of cardiac glycosides reached 2.58mg DE/gDW at day 19. Highest extracellular production was reached between day 2 and 7 (6.19mg DE/L). Highest extracellular phenolic content was 80.61 ± 5.16mg GAE/L at day 7. Intracellular phenolic content increased to 2.76 ± 0.14mg GAE/gFW at day 7 and remained constant until day 19. ROS production at day 7 could be related to sucrose and glucose total consumption. Pearson Product-Moment Correlation Coefficient (ρ) showed that the phenolic compounds in cell suspension cultures of T. peruviana were responsible for the observed anti-oxidant activity. All together, these results give the first steps in metabolic behavior in cell suspension cultures of T. peruviana.
Se establecieron cultivos en suspensión de la especie vegetal Thevetia peruviana en oscuridad, durante 19 días, para estudiar el comportamiento cinético de producción de biomasa, consumo de sustrato, producción de glicósidos cardiotónicos, polifenoles, especies reactivas de oxígeno y actividad antioxidante. Los resultados mostraron una alta producción de biomasa (18,80g PS/L), al igual que consumo total de sacarosa, a los 7 días de cultivo. Se observó un consumo preferencial de glucosa sobre fructosa durante todo el cultivo. La producción de glicósidos cardiotónicos intracelulares alcanzó valores de 2,58mg ED/g PS, al día 19. La mayor producción extracelular (6,19mg ED/L), se alcanzó entre los días 2 y 7. El mayor contenido de compuestos fenólicos extracelular fue de 80,61 ± 5,16mg GAE/L, en el día 7. El contenido de compuestos fenólicos intracelulares incrementó a 2,76 ± 0,14mg AGE/gPF, al día 7 y se mantuvo constante, hasta el día 19. La producción de EROs, al día 7, puede estar relacionada con el consumo total de sacarosa y glucosa. El coeficiente de correlación producto-momento de Pearson (ρ) indicó que los compuestos fenólicos en cultivos celulares en suspensión de T. peruviana eran los responsables de la actividad antioxidante observada. En conjunto, estos resultados brindan las primeras bases relacionadas al comportamiento metabólico de cultivos celulares en suspensión, de T. peruviana.
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OBJECTIVE@#To evaluate the impact of plant growth regulators including kinetin (KN), benzyl adenine and naphthalene acetic acid, yeast extract and casein hydrolyzate on biomass accumulation of Vietnamese ginseng Panax vietnamensis (P. vietnamensis) in cell suspension culture.@*METHODS@#Cell suspension cultures were established from friable calluses derived from leaves and petioles of 3-year-old in-vitro P. vietnamensis plants. The cell suspension cultures were grown in Murashige and Skoog basal media supplemented with various concentrations of KN, benzyl adenine, naphthalene acetic acid, and yeast extract and casein hydrolyzate.@*RESULTS@#All tested factors generated an increase in the cell biomass of P. vietnamensis in suspension culture, but the impact of each varies depended on the factor type, concentration, and incubation period. Addition of 2.0 mg/L KN resulted in the largest biomass increase after 24 d, (57.0 ± 0.9) and (3.1 ± 0.1) mg/mL fresh and dry weight, respectively, whereas addition of benzyl adenine or naphthalene acetic acid produced optimum levels of Panax cell biomass at 1.0 and 1.5 mg/L, respectively. Addition of the elicitor yeast extract led to a 1.4-2.4 fold increase in biomass of P. vietnamensis, while addition of casein hydrolyzate enhanced biomass accumulation 1.8-2.6 fold.@*CONCLUSIONS@#The addition of each factor causes significant changes in biomass accumulation of P. vietnamensis. The largest biomass accumulation is from cultures grown in MS media containing 2.0 mg/L KN for 24 d. The outcome of the present study provides new insights into the optimal suspension culture conditions for studies on the in vitro cell biomass production of P. vietnamensis.
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Objective To evaluate the impact of plant growth regulators including kinetin (KN), benzyl adenine and naphthalene acetic acid, yeast extract and casein hydrolyzate on biomass accumulation of Vietnamese ginseng Panax vietnamensis (P. vietnamensis) in cell suspension culture. Methods Cell suspension cultures were established from friable calluses derived from leaves and petioles of 3-year-old in-vitro P. vietnamensis plants. The cell suspension cultures were grown in Murashige and Skoog basal media supplemented with various concentrations of KN, benzyl adenine, naphthalene acetic acid, and yeast extract and casein hydrolyzate. Results All tested factors generated an increase in the cell biomass of P. vietnamensis in suspension culture, but the impact of each varies depended on the factor type, concentration, and incubation period. Addition of 2.0 mg/L KN resulted in the largest biomass increase after 24 d, (57.0 ± 0.9) and (3.1 ± 0.1) mg/mL fresh and dry weight, respectively, whereas addition of benzyl adenine or naphthalene acetic acid produced optimum levels of Panax cell biomass at 1.0 and 1.5 mg/L, respectively. Addition of the elicitor yeast extract led to a 1.4–2.4 fold increase in biomass of P. vietnamensis, while addition of casein hydrolyzate enhanced biomass accumulation 1.8–2.6 fold. Conclusions The addition of each factor causes significant changes in biomass accumulation of P. vietnamensis. The largest biomass accumulation is from cultures grown in MS media containing 2.0 mg/L KN for 24 d. The outcome of the present study provides new insights into the optimal suspension culture conditions for studies on the in vitro cell biomass production of P. vietnamensis.
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AIM@#To investigate the effects of feeding phenylalanine (Phe) and tyrosine (Tyr) on the accumulation of total phenolic compounds and four phenylethanoid glycosides (PeGs) to a cell suspension culture of the parasitic plant Cistanche deserticola.@*METHOD@#A cell suspension culture of C. deserticola was established and precursors of different concentrations were fed. In each group, the cell was sampled at the 24(th) day after inoculation. The content of total phenolic compounds and four PeGs compounds were determined using the Folin-Ciocalteu method and an HPLC method, respectively.@*RESULTS@#In the Phe fed cells, the maximum PeGs yield was achieved when Phe was fed at 1.5 mmol·L(-1) and the yield reached 1.13 times the control cell concentration. In the Tyr fed cells, the maximum yield of PeGs was 1.60 times of control when 0.75 mmol·L(-1) Tyr was fed to the cells. Furthermore, it was found that the salidroside yield was 4.01 times of control group when 5 mmol·L(-1) Tyr was fed.@*CONCLUSION@#Tyr is a better precursor for PeGs accumulation compared with Phe, and the rate limiting enzymes might be involved in the Tyr branch.
Subject(s)
Cell Culture Techniques , Methods , Cistanche , Chemistry , Metabolism , Culture Media , Chemistry , Metabolism , Glycosides , Metabolism , Phenylalanine , Metabolism , Tyrosine , MetabolismABSTRACT
J. curcas has been studied in different countries and some interesting agronomic, pharmacological and industrial properties have been reported. More recently, it has been considered an important alternative source for biofuel production. The objective of this study was to establish a long-term method for the maintenance of calli and cell suspension cultures of the local species J. curcas and J. gossypifolia, in order to allow future studies for novel compounds with pharmaceutical or industrial applications. For this, friable calli were successfully induced from hypocotyl segments of J. curcas and J. gossypifolia that were cultured in semisolid MS media supplemented with 1.5mg/L, and 0.5mg/L of 2,4-D, respectively. Cell suspension cultures of J. curcas were established using 1g of 35 and 60-day calli, in 50mL of liquid MS media supplied with 1.5mg/L of 2,4-D; sucrose and maltose were additionally evaluated as carbon sources. After 35 days, cell suspension cultures initiated with 35-day calli, showed greater cell growth with a maximum biomass of 194.9g/L fresh weight, 6.59g/L dry weight and 17.3% packed volume. The exponential phase ended at day 35 for cultures initiated with 35-day calli, and at day 21 for cultures initiated with 60-day calli. Higher biomass production was obtained with sucrose. Cell cultures were established with 35-day calli in MS media with the same 2,4-D concentration used for calli induction and 30g/L sucrose. This medium was considered optimum for the maintenance and growth of cell suspensions for both species, with sub-cultures every 20 days. The biotechnological potential for the production of bioactive compounds in these species for pharmacological, agricultural and industrial applications is being evaluated.
J. curcas es un importante recurso alternativo de biocombustible. Por otro lado, propiedades de interés agronómico, farmacológico e industrial han sido reportadas para esta especie. El objetivo de este estudio fue el establecimiento y mantenimiento a largo plazo de callos y cultivos celulares en suspensión de J. curcas y J. gossypifolia, con el objetivo de permitir futuros estudios para nuevos compuestos con aplicaciones farmaceúticas e industriales. Los callos friables fueron exitosamente inducidos a partir de segmentos de hipocótilos J. curcas and J. gossypifolia cultivados en medio MS semisólido suplementado con 1.5mg/L y 0.5mg/L of 2,4-D, respectivamente. Los cultivos celulares en suspensión de J. curcas fueron establecidos utilizando 1g de callos de 35 y 60 días de edad en 50mL de medio MS líquido adicionado con 1.5mg/L de 2,4-D. Después de 35 días, los cultivos en suspensión celular iniciados con callos de 35 días, mostraron mayor crecimiento celular con una biomasa máxima de 194.9g/L de peso fresco y 6.59g/L de peso seco y 17.3% de volumen empacado. La fase exponencial finalizó al día 35 en los cultivos iniciados con callos de 35 días, y al día 21 en los cultivos iniciados con callos de 60 días. Dos fuentes de carbono fueron evaluadas: sacarosa y maltosa. La producción de mayor biomasa fue obtenida con sacarosa. Los cultivos celulares se establecieron con callos de 35 días cultivados en medio MS con la misma concentración de 2,4-D utilizada para la inducción de callos y 30g/L de sacarosa. Este medio fue considerado el óptimo para el mantenimiento y crecimiento de suspensiones celulares en ambas especies con subcultivos cada 20 días. El potencial biotecnológico para la producción de compuestos bioactivos en estas especies, para aplicaciones farmacológicas, agrícolas e industriales está siendo evaluado.
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Cell Culture Techniques/methods , Jatropha/growth & development , Biomass , Jatropha/drug effects , Maltose/administration & dosage , Suspensions , Sucrose/administration & dosage , Time Factors , /administration & dosageABSTRACT
Aims: The study was conducted to develop the protocol for callus culture, cell suspension culture and to determine antibacterial activity of Ricinus communis L. cv. Roktima in cell extract. Study Design: Hypocotyl segments used as explants in callus culture and agar disk diffusion method used for antibacterial activity test. Place and Duration of Study: Institute of Biological Sciences, Rajshahi University, Rajshahi, Bangladesh during the period of 2010-2012. Methodology: MS medium supplemented with different growth regulators were used for callus induction and cell culture and paper disc diffusion method was used for the determination of antibacterial activities. Growth inhibition was determined against five gram positive bacteria viz., Sarcina lutea, Staphylococcus aureus, Bacillus megaterium, Bacillus subtillus, Bacillus halodurans, six gram negative bacteria viz., Shigella sonnei, Klebsiella species, Proteus species, Escherichia coli, Pseudomonas aeruginosa and Salmonella typhi by using disc diffusion and micro broth dilution techniques. Results: Auxins NAA, 2,4-D and IAA played a great role in callus induction but 2.0 mg/L BAP + 0.5 mg/L NAA and 2.0 mg/L BAP + 0.8 mg/L NAA concentrations proved to be most suitable combinations for induction of callus in R. communis L.cv. Roktima. Cells were cultured on the MS medium having 2.0 mg/L BAP + 0.2 mg/L NAA in which the rate of cell growth found highest and the cell continued to grow until 14 days. The peak period of cell growth was observed from 4th d to 6th d. Antimicrobial test with eleven bacteria demonstrated that the extracts of cell suspension culture of R. communis L .cv. Roktima holds the merit of antimicrobial activity and it was considered to be the potent source of antibacterial compounds and a possible source for obtaining the toxin ricin. Conclusion: In summary, the results obtained in the present investigation demonstrated that the extracts of cell suspension culture of R. communis L. cv. Roktima had the antibacterial activity and considered to be the potent source of antibacterial compounds.
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Objective:To investigate the effects of some culture conditions on production of asiaticoside from centella (Centella asiatica L. Urban) cells cultured in 5-L bioreactor. Methods: The centell cell suspension culture was conducted in 5-L bioreactor to investigate the growth and asiaticoside accumulation under various conditions. Asiaticoside content was determined by HPLC analysis. Results:The results showed that the cell growth and asiaticoside accumulation peaked after 24 d of culture at an agitation speed of 150 r/min and aeration rate of 2.5 L/min. The cell biomass reached a maximum value of 302.45 g fresh weight (31.45 g dry weight) and growth index of 3.03 with inoculum size of 100 g. However, asiaticoside content was the highest (60.08 mg/g dry weight) when culture was initiated with an inoculum size of 50 g. Conclusions: The present study found the suitable conditions for growth of centella cells and their asiaticoside production in bioreactor.
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Melastoma malabathricum, belongs to the Melastomaceae family, is an important medicinal plant widely distributed from Madagascar to Australia, that is used in traditional remedies for the treatment of variousailments. Besides its medicinal properties, it has been identified as a potential source of anthocyanin production.The present study was carried out to investigate the effect of sucrose and methyl jasmonate and feeding time oncell biomass yield and anthocyanin production in cell suspension culture of M. malabathricum. Addition of differentconcentrations of sucrose into the cell culture of M. malabathricum influenced cell biomass and pigment accumulation. The addition of methyl jasmonate was found to have no effect on cell biomass but the presence of higher amount (12.5-50mg/L) had caused a reduction in anthocyanin production and accumulation. MS medium supplemented with 30g/L sucrose and 3.5 mg/L of MeJA added on cero day and 3rd day produced high fresh cell mass at the end of nine days of culture but did not support the production of anthocyanins. However, cells cultured in the medium supplemented with 45g/L sucrose without MeJA showed the highest pigment content (0.69±0.22Cv/g-FCM). The cells cultured in MS medium supplemented with 30 g/L sucrose with 3.5mg/L MeJA added on the 3rd and 6th day of culture, showed the lowest pigment content (0.37-0.40Cv/g-FCM). This study indicated that MeJA was not necessary but sucrose was needed for the enhancement of cell growth and anthocyanin production in M. malabathricum cell cultures. Rev. Biol. Trop. 59 (2): 597-606. Epub 2011 June 01.
elastoma malabathricum pertenece a la familia de las melastomáceas, es una planta medicinal importante ampliamente distribuida desde Madagascar hasta Australia, que se utiliza en remedios tradicionales para el tratamiento de diversas dolencias. Además de sus propiedades medicinales, se ha identificado como una fuente potencial de producción de antocianinas. En esta investigación se estudió el efecto de la sucrosa, el metil jasmonato y el tiempo de ingestión en la producción de biomasa de las células y la producción de antocianinas, en el cultivo de células en suspensión de M. malabathricum. La adición de diferentes concentraciones de sucrosa al cultivo de células de M. malabathricum influencia la biomasa de las células y la acumulación de pigmento. La adición de metil jasmonato no tuvo ningún efecto sobre la biomasa celular, pero la presencia de una cantidad más alta (12.5-50mg/L) causó una reducción en la producción y acumulación de antocianinas. El medio MS complementado con sucrosa 30g/L y 3.5mg/L de MeJA en el día cero y el tercer día produjo una gran masa de células frescas al final de los nueve días de cultivo pero no se pudo mantener la producción de antocianinas. Sin embargo, las células cultivadas en el medio complementado con 45g/L de sucrosa sin MeJA mostró el mayor contenido de pigmento (0.69±0.22cv/g-fcm). Las células cultivadas en el medio MS complementado con 30 g/L de sucrosa y con 3.5 mg/l MeJA en el tercer y sexto día de cultivo, mostró el menor contenido de pigmentos (0.37-0.40cv/g-fcm). Este estudio indicó que MeJA no era necesario pero la sucrosa sí se necesitaba para mejorar el crecimiento celular y la producción de antocianinas en cultivos de células de M. malabathricum.
Subject(s)
Acetates , Anthocyanins/biosynthesis , Biomass , Cyclopentanes/pharmacology , Melastomataceae/drug effects , Oxylipins/pharmacology , Sucrose/pharmacology , Cells, Cultured , Melastomataceae/growth & development , Melastomataceae/metabolismABSTRACT
Objective: Single-cell of Dendrobium huoshanenes was cultured by using cell suspension culture for solving the problem of resource shortage of medicinal plant D. huoshanenes. Methods: The single-cell of D. huoshanenes was obtained by different preparing ways, effects of various factors on cell growth were studied and the optimal growing conditions of cell suspension culture were investigated by orthogonal design. Results: Through the orthogonal test, the results indicated that the optimum conditions for suspension culture of D. huoshanenes cell were as follow: is that the concentration of sucrose was 35 g/L, NH4NO3 was 1.3 g/L, pH value was 5.6, the hormone IAA was 0.4 mg/L , 6-BA was 1.0 mg/L, the best culture time was 14-18 d. Under optimal culture conditions, the concnentration of single cells in this test was 9.42 X 105 cell/mL. Conclusion: After four successive subculture, cellular morphology of D. huoshanenes shows circular shape or elliptic shape and the chip of cell is decreased. The higher concentration of D. huoshanenes cell could be obtained by using the optimal growing conditions of cell suspension culture.
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AIM: To compare cell-suspension and explant culture of mouse corneal epithelial cells (MCEC).METHODS: MCEC were cultured by cell-suspension culture and explant culture, respectively. Colony forming efficiency (CFE) and cell proliferation were determined. The expression of corneal epithelial progenitor cell marker p63 and K19, as well as differentiation marker K12 was investigated by Western blotting.RESULTS: Twenty of 25 (80％) cornea explant were successfully subcultured to passage1 (P1), while only 12％ cell-suspension culture were successfully subcultured to P1. There were statistical significance between explant culture and cell-suspension culture (P<0.01). Up to 55％ of P1 cells in explant culture were passaged over P10 and were stably subcultured though at least 25 passages. However, cells cultured in suspension culture never achieved confluence in P2. CEF of P1 in explant culture was higher than P1 in cell-suspension culture (P=0.02) and CEF of P20 in explant culture was higher than P1 in explant (P=0.001). Immunostaining images showed expression of p63 and K19 in cell-suspension culture P1 and explant culture P1 and P20. K12 was expressed in P1 of both cell-suspension culture and explant culture, however, there was not K12 expressed in P20 of explant culture.CONCLUSION: In MECE culture, compared with cell-suspension culture, the explant culture is a preferable option.
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The type of basic media and the contents of plant growth substances were investigated by orthogonal design experiment,and also the effects of different culture conditions on the growth of suspension cells and the accumulation of total flavonoids in Eucommia ulmoides were studied.The results showed that B5 medium supplemented with 0.5mg/L NAA,0.6mg/L 6-BA and 30g/L sucrose,at initial pH 5.0～5.5,20g(FW)/L inoculation quantity and 110 r/min of rotation speed was a preferable culture conditions for E.ulmoides suspension cells growth and flavonoids synthesis.The results of metabolic kinetics analysis for E.ulmoides cell suspension culture showed that the logistic and Luedeking-Piret equations can be used for describing the kinetics of cell growth,sucrose consumption and flavonoids production during the process.The maximum specific growth rate(?m),the actual growth yield based on sucrose(YG) and maintenance coefficient(m) were 0.417/d,0.619g/g and 0.0206g/(g?d-1) respectively.All these outcomes could give a basis for establishing the suspension cell culture of E.ulmoides and production of the natural active components in large-scale.
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Hormones are the main materials which regulate the plant growth and the secondary metabolites formation of plant. The effects of hormones on cell biomass and metabolites content and the application progress of honrmones in plant cell suspension culture is reviewed. It covers the influence of exogenous hormones’ categories, concentration and proportion on cell biomass and metabolites content in plant cell suspension culture, the developmental course of the endogenous hormones determination, the changes of endogenous hormones content and its effects on the cell biomass and metabolites content, the relation of interaction between exogenous hormones and endogenous hormones, the study on the effect of new hormone varieties on the culture.
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Objective To investigate the biotransformation of podophyllotoxin by cell suspension culture and root culture systems of Rheum palmatum.Methods Plant tissue culture technology was employed.The products were isolated by silica gel column chromatography and their structures were elucidated by spectral methods.Results Adding of podophyllotoxin had no obvious effects on the pH value of cell suspension culture and root culture systems of R.palmatum.In the case of present study,73.8% of podophyllotoxin was converted by cell suspension culture and 56.3% by root culture.The products were different between the two culture system.Podophyllotoxin was converted into picropodophyllin with cell suspension culture system and many other products besides epipodophyllotoxin with root culture system.Conclusion Both of cell suspension culture and root culture systems of R.palmatum are able to convert podophyllotoxin.
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Object To investigate the biotransformation of cantharidin by cell suspension cultures of Platycodon grandiflorus (Jacq ) A DC Methods Cantharidin was added into cell suspension cultures of P grandiflorus and incubated for another six days The culture supernatant was extracted with EtOAc and then subjected to silica gel chromatography Results Two products were isolated as a mixture in a molar ratio of about 2∶1 Their structures were identified on the basis of spectroscopic data as 1? OH cantharidin (Ⅱa) and 1? OH cantharidin (Ⅱb) The relative configuration was elucidated according to NOESY spectrum Conclusion The two products were new cantharidin derivatives