To date enormous attempts have been devoted to improve Taxol production exploiting various methodologies from bioprocess engineering to biotechnological and synthetic approaches. We have developed a 2-stage suspension cell culture of Taxus baccata L. using modified B5 medium in order to improve cell growth as well as productivity. After callus induction and cell line selection, B5 medium was supplemented with vanadyl sulfate (0.1 mg/l), silver nitrate (0.3 mg/l) and cobalt chloride (0.25 mg/l) at the first day of stage I culture to maximize cell growth. This medium was further supplemented with sucrose (1%) and ammonium citrate (50 mg/l) on day 10 and sucrose (1%) and phenylalanine (0.1 mM) on day 20 (i.e., biomass growth medium). At stage II (day 25), two different concentrations of several elicitors such as methyl jasmonate (10 or 20 mg/l), salicylic acid (50 or 100 mg/l) and fungal elicitor (25 or 50 mg/l) were added to the biomass growth medium with the aim of improving cellular productivity. For morphological analysis, microscopic inspection was carried out during cultivation. Cell-associated and extracellular amount of Taxol were detected and measured using HPLC methodology. At stage I, overall Taxol amount of biomass growth medium was 13.75 mg/l (i.e., 5.6-fold higher than that of untreated B5 control). At stage II, treated cells with methyl jasmonate (10 mg/l), salicylic acid (100 mg/l) and fungal elicitor (25 mg/l) produced the highest amount of Taxol (39.5 mg/l), which is 16-fold higher than that of untreated B5 control (2.45 mg/l). Microscopic analyses of Taxus cells in suspension cultures showed various positional auto-fluorescence showing direct correlation with Taxol production. Our studies revealed that intervallic supplementation of B5 medium with combination of biomass growth factors at stage I and mixture of elicitors at stage II could significantly increase Taxol production. Thus, we suggest that the exploitation of this methodology may improve the production of Taxol since demands for Taxol pharmaceuticals are increasingly growing and resource paucities have limited its direct harvesting from Taxus trees.
The search for natural antioxidants, especially of plant origin, has notably increased in recent years. Bunium persicum Boiss. is an economically important medicinal plant growing wild in the dry temperature regions in Iran. In this study, chemical constituents of the essential oil of the seed from Bunium persicum Boiss. have been studied by GC/MS technique. The major components were caryophyllene (27.81%), gamma-terpinene (15.19%), cuminyl acetate (14.67%). Individual antioxidant assays such as, DPPH* scavenging activity and beta-carotene bleaching have been carried out. In DPPH* system, the EC(50) value of essential oil was determined as 0.88 mg/mL. In beta-carotene bleaching antioxidant activity of essential oil (0.45%) was almost equal to BHT at 0.01%. In addition, the antioxidant activity of the essential oil was evaluated in crude soybean oil by monitoring peroxide and thiobarbituric acid values of the oil substrate. The results showed that the Bunium persicum essential oil (BPEO) was able to reduce the oxidation rate of the soybean oil in the accelerated condition at 60 degrees C (oven test). The essential oil at 0.06% showed the same effect of BHA at 0.02%. Hence, BPEO could be used as an additive in food after screening.
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