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Jian Zaiyou Meng Li Xu Guifang Zhou Xiuren 《Journal of industrial microbiology & biotechnology》2013,40(11):1297-1302
The objective of this study was to isolate endophytic fungi producing baccatin III from yew for the purpose of baccatin III and paclitaxel manufacture. Surface sterilized bark of Taxus wallichiana var. mairei was used as source material with potato dextrose agar culture medium for isolation of endophytic fungi. Fungal cultures were extracted with a mixture of chloroform/methanol (1:1, v/v) and the baccatin III in the extracts was determined and authenticated with LC–MS. An endophytic fungus that produced baccatin III was identified by ITS rDNA and 26S D1/D2 rDNA sequencing. A total of 192 endophytic fungal strains were isolated from T. wallichiana var. mairei. Only one of the 192 strains produced baccatin III and it was identified as Diaporthe phaseolorum. The productivity of this strain cultured in PDA culture medium was 0.219 mg/l. The isolated endophytic fungus produced baccatin III at a relatively high level and shows promise as a producing strain for baccatin III and paclitaxel manufacture after strain improvement. 相似文献
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M. Bonfill S. Bentebibel E. Moyano J. Palazón R. M. Cusidó R. Eibl M. T. Piñol 《Biologia Plantarum》2007,51(4):647-652
The effects of 100 and 200 μM methyl jasmonate (MJA) on cell proliferation and paclitaxel and baccatin III production were
investigated in free and alginate immobilized cells of Taxus baccata growing in a selected product formation culture medium. The greatest accumulation of paclitaxel (13.20 mg dm−3) and baccatin III (4.62 mg dm−3) occurred when 100 μM MJA was added to the culture medium of cells entrapped using a 1.5 and 2.5 % alginate solution. The
effects of different treatments on the viability of cultured cells and their capacity to excrete both taxanes into the surrounding
medium were considered. 相似文献
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Miriam Onrubia Elisabet Moyano Mercedes Bonfill Rosa Mª Cusidó Alain Goossens Javier Palazón 《Journal of plant physiology》2013
Coronatine is a toxin produced by the pathogen Pseudomonas syringae. This compound has received much attention recently for its potential to act as a plant growth regulator and elicitor of plant secondary metabolism. To gain more insight into the mechanism by which elicitors can affect the biosynthesis of paclitaxel (Px) and related taxanes, the effect of coronatine (Cor) and methyl jasmonate (MeJA) on Taxus media cell cultures has been studied. For this study, a two-stage cell culture was established, in which cells were first cultured for 14 days in a medium optimised for growth, after which the cells were transferred to medium optimised for secondary metabolite production. The two elicitors were added to the medium at the beginning of the second stage. Total taxane production in the cell suspension was significantly enhanced by both elicitors, increasing from a maximum level of 8.14 mg/L in control conditions to 21.48 mg/L (day 12) with MeJA and 77.46 mg/L (day 16) with Cor. Expression analysis indicated that the txs, t13oh, t2oh, t7oh, dbat, pam, bata and dbtnbt genes were variably induced by the presence of the elicitors. Genes encoding enzymes involved in the formation of the polihydroxylated hypothetical intermediate (TXS, T13OH, T2OH, T7OH) and the phenylalanoil CoA chain (PAM) were stronger induced than those encoding enzymes catalysing the last steps of the Px biosynthetic pathway (DBAT, BAPT and DBTNBT). Notably, although taxane accumulation differed qualitatively and quantitatively following MeJA- or Cor-elicitation, gene expression induction patterns were similar, inferring that both elicitors may involve distinct but yet uncharacterised regulatory mechanisms. 相似文献
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To achieve a better understanding of the metabolism and accumulation of paclitaxel and baccatin III in cell cultures of Taxus, inhibitors of the early steps in the terpenoid pathway were applied to a cell suspension culture of Taxus baccata: fosmidomycin as an inhibitor of the non-mevalonate branch of the pathway, and mevinolin as an inhibitor of the mevalonate branch. Synthesis of both taxanes in the cell suspension was first increased when cultured in the product formation medium supplemented with methyljasmonate (100 microM). The product formation medium was selected after assaying 24 different culture media. When fosmidomycin (200 microM) was added to the product formation medium together with the elicitor, the accumulation of paclitaxel and baccatin III was reduced by up to 3.0 and 1.5 times, respectively, whereas the inhibitory effect of mevinolin (1 microM) was only clearly exerted in the case of paclitaxel. Under the conditions of our experiment, we conclude that in the synthesis of both taxanes, the non-mevalonate pathway is the main source of the universal terpenoid precursor isopentenyl diphosphate (IPP). 相似文献
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Nikolakakis A Caron G Cherestes A Sauriol F Mamer O Zamir LO 《Bioorganic & medicinal chemistry》2000,8(6):1269-1280
An efficient conversion of Taxus canadensis abundant taxane, 9-dihydro-13-acetylbaccatin III to baccatin III is described. Since the synthesis of paclitaxel from baccatin III has been reported, this work can be used for additional supply of this powerful anticancer drug. In addition, new taxanes derived from skeletal rearrangements originating from oxidation reduction reactions of the Canadian yew major taxane, are reported. 相似文献
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A new and simple method was developed to recover paclitaxel from the extracellular culture medium of Taxus chinensis. More than 80% of paclitaxel in the medium was obtained by adding 7.8 mM MgSO4 or greater and then centrifuging. The concentration of baccatin III in the supernatant did not change after MgSO4 treatment while paclitaxel was precipitated in the pellet. This method was used to recover paclitaxel without baccatin III from the extracellular culture medium of Taxus species. 相似文献
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Bentebibel S Moyano E Palazón J Cusidó RM Bonfill M Eibl R Piñol MT 《Biotechnology and bioengineering》2005,89(6):647-655
Paclitaxel and baccatin III-producing cells of Taxus baccata were immobilized within Ca(2+)-alginate beads. Under established optimum conditions for the biosynthesis of both taxanes, the yields of paclitaxel and baccatin III in shake-flask cultures of free cells increased by factors of up to 3 and 2, respectively, in the corresponding cultures of immobilized cells. Although the scale-up from shake-flask to bioreactor culture usually results in reduced productivities when both free and immobilized cells were grown in the same optimum conditions in three different bioreactor types (Stirred, Airlift, and Wave) running for 24 days in a batch mode and with the system optimized in each case, there was a considerable increase in the yields of paclitaxel and baccatin III. Among the reactors, the Stirred bioreactor was the most efficient in promoting immobilized cell production of paclitaxel, giving a content of 43.43 mg.L(-1) at 16 days of culture, equivalent to a rate of 2.71 mg.L(-1).day(-1). To our knowledge, the paclitaxel productivity obtained in this study is one of the highest reported so far by academic laboratories for Taxus species cultures in bioreactors. 相似文献
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Yamamoto N Mukai Y Matsuoka M Kano-Murakami Y Tanaka Y Ohashi Y Ozeki Y Odani K 《Plant physiology》1991,95(2):379-383
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