• Title/Summary/Keyword: taxol production

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Effects of Inoculum Density and Basal Media on Cell Growth and Taxol Production in Taxus Cell Suspension Cultures (주목 세포배양에서 초기 접종농도와 기본배지가 세포증식과 Taxol 생산에 미치는 영향)

  • 황용순;김석우
    • KSBB Journal
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    • v.11 no.5
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    • pp.600-605
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    • 1996
  • Optimum inoculum concentration for the production of taxol was determined in Taxus brevifolia and Taxus cuspidata cell suspension cultures. By fresh weight, 2.5, 5, 7.5, 10 g/flask of cells were inoculated and cell growth as well as taxol production were examined. In both Taxus cell cultures, the higher the inoculum concentration, the shorter the length of the lag period. The optimum inoculum concentration for taxol production was found to be 5 g/flask. To produce taxol in large quantity, utilization of proper medium was thought to be important. In case of using a production medium with 6% sucrose, taxol production was noticed. Its level reached the maximum at the 9th day of culture and decreased afterwards. However, taxol was not detected from cell cultures in growth medium.

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Effect of Nitrogen, Phosphate and Cell Immobilization on Taxol Production from Cell Cultures of Taxus cuspidata (주목 (Taxus cuspidata) 세포배양에서 질소원, 인산, 세포고정화가 Taxol 생산에 미치는 영향)

  • Park, Jong-Hwa;Chung, In-Sik
    • Applied Biological Chemistry
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    • v.38 no.4
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    • pp.308-312
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    • 1995
  • The effects of nitrogen, phosphate in modified B5 medium and cell immobilization on cell growth and taxol production were investigated using cell cultures of Taxus cuspidata. The ratio of nitrate to ammonium was found to be an important parameter. The ratio of 1 increased taxol production 10-fold, compared to the original ratio of 20 in modified B5 medium. Reducing phosphate concentration inhibited cell growth, but increased taxol production noticeably. Immobilized cells produced a taxol concentration of ${\sim}120\;g/l$

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Taxol Production by an Endophytic Fungus, Fusarium redolens, Isolated from Himalayan Yew

  • Garyali, Sanjog;Kumar, Anil;Reddy, M. Sudhakara
    • Journal of Microbiology and Biotechnology
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    • v.23 no.10
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    • pp.1372-1380
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    • 2013
  • Different endophytic fungi isolated from Himalayan Yew plants were tested for their ability to produce taxol. The BAPT gene (C-13 phenylpropanoid side chain-CoA acetyl transferase) involved in the taxol biosynthetic pathway was used as a molecular marker to screen taxol-producing endophytic fungi. Taxol extracted from fungal strain TBPJ-B was identified by HPLC and MS analysis. Strain TBPJ-B was identified as Fusarium redolens based on the morphology and internal transcribed spacer region of nrDNA analysis. HPLC quantification of fungal taxol showed that F. redolens was capable of producing $66{\mu}g/l$ of taxol in fermentation broth. The antitumour activity of the fungal taxol was tested by potato disc tumor induction assay using Agrobacterium tumefaciens as the tumor induction agent. The present study results showed that PCR amplification of genes involved in taxol biosynthesis is an efficient and reliable method for prescreening taxol-producing fungi. We are reporting for the first time the production of taxol by F. redolens from Taxus baccata L. subsp. wallichiana (Zucc.) Pilger. This study offers important information and a new source for the production of the important anticancer drug taxol by endophytic fungus fermentation.

Isolation and Identification of Taxol, an Anticancer Drug from Phyllosticta melochiae Yates, an Endophytic Fungus of Melochia corchorifolia L.

  • Kumaran, Rangarajulu Senthil;Muthumary, Johnpaul;Hur, Byung-Ki
    • Food Science and Biotechnology
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    • v.17 no.6
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    • pp.1246-1253
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    • 2008
  • Phyllosticta melochiae, an endophytic fungus isolated from the healthy leaves of Melochia corchorifolia, was screened for the production of an anticancer drug, taxol on modified liquid medium and potato dextrose broth medium in culture for the first time. The presence of taxol was confirmed by spectroscopic and chromatographic methods of analysis. The amount of taxol produced by this fungus was quantified by high performance liquid chromatography. The maximum amount of fungal taxol production was recorded as $274{\mu}g/L$. The production rate was increased to $5.5{\times}1,000$ fold than that found in the culture broth of earlier reported fungus, Taxomyces andreanae. The fungal taxol extracted also showed a strong cytotoxic activity in the in vitro culture of tested human cancer cells by apoptotic assay. The results designate that the fungal endophyte, P. melochiae is an excellent candidate for an alternate source of taxol supply and can serve as a potential species for genetic engineering to enhance the production of taxol to a higher level.

Taxol Production in Taxus Cell Cultures: Effects of Various Elicitors (주목세포배양에 의한 Taxol 생산: 여러 가지 Elicitor가 미치는 영향)

  • 윤정환;김진훈
    • KSBB Journal
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    • v.10 no.2
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    • pp.143-148
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    • 1995
  • The effects of various elicitors, metabolic inhibitors and growth regulators on the production of diterpenoid anticancer agent taxol were investigated in cell suspension cultures of Taxus brevifolia. Cell cultures of T. brevifolia were treated by 5 kinds of biotic elicitors, 5 kinds of abiotic elicitors, 2 kinds of metabolic inhibitors and 8 kinds of growth regulators at the end of exponential growth phase. Among those treatments, chlorocholine chloride-an inhibitor of plant steroid metabolism-increased the taxol production most significantly. From a series of optimization studies, it was found that the addition of 1mM of chlorocholine chloride at the 9th day of culture was the best for taxol production. Taxol yield under this condition was 0.72mg/$\ell$.

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Effects of Precursors and Exogenous Taxanes on Taxane Production by Cell Suspension Cultures (전구체 및 외부공급 Taxane이 세포배양에 의한 Taxane 생산에 미치는 영향)

  • 황용순;김진우
    • KSBB Journal
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    • v.11 no.3
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    • pp.323-328
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    • 1996
  • Effects of three kinds of precursors as well as exogenous taxanes on the production of taxol and its derivatives were investigated in Taxus cuspidata and Taxus brevifolia cell suspension cultures. When geraniol was added as a precursor of geranylgeranyl pyrophosphate to enhance diterpenoid metabolism, production of some taxanes including taxol was enhanced. The time of addition and amount of feeding were found to be important factors. Feeding of camphor and menthol resulted in negative effects on taxol production. Influences of exogenous taxanes on taxane production by cell cultures were found to be very complicated. When taxol, baccatin III, cephalomannine were exogenously added into the culture, production of baccatin III, 7-epi-10-deacetyltaxol, 10-deacetyltaxol was increased respectively.

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Systematic Analysis of the Anticancer Agent Taxol-Producing Capacity in Colletotrichum Species and Use of the Species for Taxol Production

  • Choi, Jinhee;Park, Jae Gyu;Ali, Md. Sarafat;Choi, Seong-Jin;Baek, Kwang-Hyun
    • Mycobiology
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    • v.44 no.2
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    • pp.105-111
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    • 2016
  • Paclitaxel (taxol) has long been used as a potent anticancer agent for the treatment of many cancers. Ever since the fungal species Taxomyces andreanae was first shown to produce taxol in 1993, many endophytic fungal species have been recognized as taxol accumulators. In this study, we analyzed the taxol-producing capacity of different Colletotrichum spp. to determine the distribution of a taxol biosynthetic gene within this genus. Distribution of the taxadiene synthase (TS) gene, which cyclizes geranylgeranyl diphosphate to produce taxadiene, was analyzed in 12 Colletotrichum spp., of which 8 were found to contain the unique skeletal core structure of paclitaxel. However, distribution of the gene was not limited to closely related species. The production of taxol by Colletotrichum dematium, which causes pepper anthracnose, depended on the method in which the fungus was stored, with the highest production being in samples stored under mineral oil. Based on its distribution among Colletotrichum spp., the TS gene was either integrated into or deleted from the bacterial genome in a species-specific manner. In addition to their taxol-producing capacity, the simple genome structure and easy gene manipulation of these endophytic fungal species make them valuable resources for identifying genes in the taxol biosynthetic pathway.

Isolation, Purification, and Identification of Taxol and Related Taxanes from Taxol-Producing Fungus Aspergillus niger subsp. taxi

  • Li, Dan;Fu, Dongwei;Zhang, Yue;Ma, Xueling;Gao, Liguo;Wang, Xiaohua;Zhou, Dongpo;Zhao, Kai
    • Journal of Microbiology and Biotechnology
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    • v.27 no.8
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    • pp.1379-1385
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    • 2017
  • The content of taxol in the bark of yews is very low, and this is not affordable from the environmental point of view. Thus, it is a necessity to look for alternative sources of taxol production to solve its supply. Currently, a large portion of the taxol in the market comes from chemical semi-synthesis, but the semi-synthetic precursors such as baccatin III and 10-deacetyl-baccatin III are extracted from needles and twigs of yew trees. Taxol-producing fungi as a renewable resource is a very promising way to increase the scale of taxol production. Our group has obtained a taxol-producing endophytic fungus, Aspergillus niger subsp. taxi HD86-9, to examine if A. niger can produce the taxanes. Six compounds from the fermentation broth of strain HD86-9 were isolated and identified by $^1H$ NMR, $^{13}C$ NMR, and ESI-MS. The results showed that the six compounds included four taxane diterpenoids (taxol, cephalomannine, baccatin III, and 10-deacetyl-baccatin III) and two non-taxane compounds (${\beta}-sitosterol$ and flavonoid isovitexin). The study verified that the taxanes can be produced by the A. niger, which is very important to taxol production via chemical semi-synthesis. Additionally, the finding is potentially very significant to solve the taxol semi-synthetic precursors extracted from needles and twigs of yew trees, and the precursor production can be easily increased through the culture condition optimization, genetic breeding, and metabolic engineering of the A. niger.

Production, Purification, and Characterization of Taxol and 10-DABIII from a new Endophytic Fungus Gliocladium sp. Isolated from the Indian Yew Tree, Taxus baccata

  • Sreekanth, D.;Syed, A.;Sarkar, S.;Sarkar, D.;Santhakumari, B.;Ahmad, A.;Khan, M.I.
    • Journal of Microbiology and Biotechnology
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    • v.19 no.11
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    • pp.1342-1347
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    • 2009
  • We have isolated endophytic fungi from the Indian yew tree, Taxus baccata, and then screened for taxol production. Out of the 40 fungal cultures screened, one fungus Gliocladium sp. was found to produce taxol and 10-DABIII (10-deacetyl baccatin III). These compounds were purified by TLC and HPLC and characterized using UV-spectroscopy, ESI-MS, MS/MS, and proton NMR. One liter of Gliocladium sp. culture yielded $10\;{\mu}g$ of taxol and $65\;{\mu}g$ of 10-DABIII. The purified taxol from the fungus showed cytotoxicity towards cancer lines HL-60 (leukemia), A431 (epidermal carcinoma), and MCF-7 (breast cancer).

Effects of taxol and ionizing radiation on cytotoxicity and prostaglandin production in KB, RPMI-2650, SW-13 and L929 (수종과 암세포주와 섬유모세포주에서 taxol과 전리방사선이 세포독성과 prostaglandin생성에 미치는 영향)

  • Lee Keon-Il;You Dong-Soo
    • Journal of Korean Academy of Oral and Maxillofacial Radiology
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    • v.28 no.1
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    • pp.127-143
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    • 1998
  • The author evaluated the effects of taxol, a microtubular inhibitor, as a possible radiation sensitizer and the production of prostaglandins on three human cancer cell lines(KB, RPMI-2650 and SW-13) and one murine cell line(L929). Each cell line was divided into four groups (control, taxol only, radiation only and combination of taxol and radiation). The treatment consisted of a single irradiation of 10Gy and graded doses (5, 50, 100, 200, 300, 500 nM) of taxol for a 24-h period. The cytotoxicity of taxol alone was measured at 1 day after(1-day group) and 4 days after(4-day group) the treatment. The survival ratio of cell was analyzed by MTT (3-(4,5-dimethylthiazol-2-yl) -2,5-dimethyl tetrazolium bromide) test. Prostaglandins(PGE2 and PGI2) were measured in the culture medium by a radioimmunoassay. The results obtained were as follows. 1. There was a significantly increased cytotoxicity of KB cells in 4-day group than those in I-day group. There was a high correlation between doses of taxol and cell viability in both groups(l-day group R=0.82741, 4-day group R=0.84655). 2. There was a significantly increased cytotoxicity of RPMI -2650 cells treated with high concentration of taxol in 4-day group than those in I-day group. Also there was a high correlation between doses of taxol and cell viability in 4-day group(R=0.93917). 3. There was a significantly increased cytotoxicity of SW-13 cells treated with high concentration of taxol in 4-day group than those in 1-day group. However no high correlation was observed between doses of taxol and cell viability in both groups(1-day group R=0.46362, 4-day group R=0.65425). 4. There was a significantly increased cytotoxicity of L929 cells treated with low concentration of taxol in 4-day group than those in 1-day group. At the same time, there was a low correlation between doses of taxol and cell viability in both groups(1-day group R=0.34237, 4-day group R=0.23381). 5. In I-day group of L929 cells, higher cytotoxicities were observed in the groups treated with 500 nM taxol than given 10 Gy radiation alone. L929 cells in I-day group alone showed a radiosensitizing effect by taxol.. 6. In addition to L929 cells, all cancer cells treated with a combination of taxol and radiation in 4-day group appeared to have some fragmented nuclei and to float on the medium. In addition, L929 cells appeared to be more confluent. 7. The level of PGE2 production was the highest in the contol KB cells. This appeared to increase in every experimental group of all three cancer cells except L929 cells. There was a significantly increased production of PGE2 in SW -13 cells treated with a combination taxol and radiation compared to the other experimental groups. 8. The level of PGE2 production in the control group of RPMI-Z650 cells was the highest. This appeared to increase in every experimental group of all cells except in SW-13 cells. This also increased significantly in RPMI-2650 cells treated with a combination of taxol and radiation compared to the other experimental groups.

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