Eurycoma longifolia is an important rare medicinal plant that contains valuable bioactive compounds. In the present study, cell suspension culture of E. longifolia was established for the production of biomass and phenolic compounds. Various medium parameters, such as concentration of auxin, salt st...
Eurycoma longifolia is an important rare medicinal plant that contains valuable bioactive compounds. In the present study, cell suspension culture of E. longifolia was established for the production of biomass and phenolic compounds. Various medium parameters, such as concentration of auxin, salt strength of the medium, and sucrose and nitrogen concentrations, were optimized for the production of biomass at the flask-scale level. Full strength Murashige and Skoog (MS) medium supplemented with $3.0mg{\cdot}L^{-1}$ naphthaleneacetic acid (NAA), 3% (w/v) sucrose, 0:60 $NH{_4}^+:NO{_3}^-$ was found suitable for biomass accumulation. Based on the optimized flask-scale parameters, cell suspension cultures were established in balloon-type bubble bioreactors, and bioprocess parameters such as inoculum density and aeration rate were optimized. Inoculum density of $50g{\cdot}L^{-1}$ and increasing aeration rate from 0.05 to 0.3 vvm, with increases every 7 days, were suitable for the accumulation of both biomass and phenolic compounds. With the optimized conditions, $14.70g{\cdot}L^{-1}$ dry biomass, $10.33mg{\cdot}g^{-1}$ DW of phenolics and $3.89mg{\cdot}g^{-1}$ DW of flavonoids could be achieved. Phenolics isolated from the cell biomass showed optimal free radical scavenging activity.
Eurycoma longifolia is an important rare medicinal plant that contains valuable bioactive compounds. In the present study, cell suspension culture of E. longifolia was established for the production of biomass and phenolic compounds. Various medium parameters, such as concentration of auxin, salt strength of the medium, and sucrose and nitrogen concentrations, were optimized for the production of biomass at the flask-scale level. Full strength Murashige and Skoog (MS) medium supplemented with $3.0mg{\cdot}L^{-1}$ naphthaleneacetic acid (NAA), 3% (w/v) sucrose, 0:60 $NH{_4}^+:NO{_3}^-$ was found suitable for biomass accumulation. Based on the optimized flask-scale parameters, cell suspension cultures were established in balloon-type bubble bioreactors, and bioprocess parameters such as inoculum density and aeration rate were optimized. Inoculum density of $50g{\cdot}L^{-1}$ and increasing aeration rate from 0.05 to 0.3 vvm, with increases every 7 days, were suitable for the accumulation of both biomass and phenolic compounds. With the optimized conditions, $14.70g{\cdot}L^{-1}$ dry biomass, $10.33mg{\cdot}g^{-1}$ DW of phenolics and $3.89mg{\cdot}g^{-1}$ DW of flavonoids could be achieved. Phenolics isolated from the cell biomass showed optimal free radical scavenging activity.
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문제 정의
Production of biomass and bioactive compounds through cell cultures can meet the demand of pharmaceutical industry and also accounts for little ease on natural stand. Therefore, major objective of present study was to establish cell suspension cultures using bioreactor system. Initially we established cell suspension cultures in Erlenmeyer flasks and optimized the various chemical parameters such as growth regulators, medium salt strength, sucrose and nitrogen concentration for accumulation biomass.
제안 방법
In the first set of experiments, the effect of indole-3-butyric acid (IBA) and naphthalene acetic acid (NAA) in the range of 0.5, 1.0, 3.0, 5.0 and 7.0 mg･L-1 were tested on biomass accumulation. In another set of experiments, the effects of MS salt strength ranging 0.
0.5 g powdered material was placed in a round bottom flask and extraction was carried out at 80 ± 1ºC for 1 h using 50 mL of distilled water, 40, 60, 80 and 100% of methanol and ethanol to find out the suitability of solvent for extraction of total phenolics and flavonoids.
The effect of nitrogen concentration [NH4+ :NO3; 0:30, 5:25, 10:20, 15:15, 20:10, 25:5 and 30:0 mM] was tested for the production of biomass in the fourth set of experiments.
대상 데이터
All experiments were set up in a completely randomized design and the data were collected from three replicates. Mean values were subjected to Duncan’s multiple range test using SPSS software (version 9.
데이터처리
Mean values were subjected to Duncan’s multiple range test using SPSS software (version 9.0).
성능/효과
All the bioreactors were maintained for 30 days at 25 ± 1ºC, with a 16 h (40 µmol･m-2･s-1) light and 8 h dark photoperiod cycle provided by 40-W white fluorescent tubes. After 30 days, growth parameters such as fresh weight, dry weight and growth ratio and the content of bioactive compounds polyphenolics and flavonoids were assessed.
To understand the growth characteristics of suspended cells of E. longifolia in shake flasks, the effect of NAA and IBA was tested and the results revealed that supplementation of NAA was superior in terms of biomass accumulation and the medium supplemented with 3.0 mg･L-1 NAA yielded optimum results (Table 1).
Plant cell cultures are usually grown heterotrophically using simple sugars and carbon sources and the levels of sucrose has been shown to affect the biomass accumulation during suspension cultures (Ramachandra Rao and Ravishankar, 2002). The results of present work showed that 3% sucrose (w/v) was found suitable for biomass accumulation in cell suspension cultures of E. longifolia (Table 3). Similar to the current results, 3% sucrose was found suitable for biomass accumulation in cell suspension cultures of Gymnema sylvestre (Nagella et al.
The results of current studies showed that 0:60 mM NH4:NO3 was responsible for highest biomass accumulation (115.4 g･L-1 fresh biomass and 6.5 g･L-1 dry biomass) among the varied concentration of NH4:NO3 tested (Table 4).
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