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1.
Rangarajulu Senthil Kumaran Johnpaul Muthumary Byung-Ki Hur 《Journal of microbiology (Seoul, Korea)》2009,47(1):40-49
Phyllosticta tabernaemontanae, a leaf spot fungus isolated from the diseased leaves of Wrightia tinctoria, showed the production of taxol, an anticancer drug, on modified liquid medium (MID) and potato dextrose broth (PDB) 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 using high performance liquid chromatography (HPLC). The maximum
amount of taxol production was recorded in the fungus grown on MID medium (461 μg/L) followed by PDB medium (150 μg/L). The
production rate was increased to 9.2 × 103 fold than that found in the culture broth of earlier reported fungus, Taxomyces andreanae. The results designate that P. tabernaemontanae is an excellent candidate for taxol production. The fungal taxol extracted also showed a strong cytotoxic activity in the
in vitro culture of tested human cancer cells by apoptotic assay. 相似文献
2.
X. Zhou Z. Wang K. Jiang Y. Wei J. Lin X. Sun K. Tang 《Applied Biochemistry and Microbiology》2007,43(4):439-443
A total of 38 endophytic fungus strains were isolated from Taxus chinensis var. mairei by the aseptic technique. Genomic DNA was extracted from isolated endophytic fungi and subjected to polymerase chain reaction
(PCR) analysis for the presence of the Taxus taxadiene synthase (TS) gene, a rate-limiting enzyme gene in the taxol biosynthetic pathway. Twelve out of 38 isolated endophytic fungus strains
showed PCR positive for the ts gene. Subsequently, taxol and its related compounds were extracted from culture filtrates and mycelia of the PCR positive
strains, separated by column chromatography, and analyzed by High Performance Liquid Chromatography and Mass Spectrum. The
analysis result showed that 3 strains could produce taxol and its related compounds at the detectible level. This study indicates
that molecular detection of the ts gene is an efficient method for primary screening of taxol or its related compound-producing endophytic fungi, which can
improve prominently screening efficiency.
Published in Russian in Prikladnaya Biokhimiya i Mikrobiologiya, 2007, Vol. 43, No. 4, pp. 490–494.
The text was submitted by the authors in English. 相似文献
3.
Taxol is an important anticancer drug widely used in the clinic. An endophytic fungus Bartalinia robillardoides (strain AMB-9) was isolated from Aegle marmelos, a medicinal plant and screened for taxol production. The fungus was identified based on the morphology of the fungal culture
and the characteristics of the spores. This fungus was grown in MID liquid medium and analyzed chromatographically and spectrometrically,
for the presence of Taxol. The amount of taxol produced by this endophytic fungus was quantified by HPLC. It produced 187.6 μg/L
of taxol which suggests that the fungus can serve as a potential material for genetic engineering to improve the production
of Taxol. This fungal taxol isolated from the organic extract of this fungal culture, has strong cytotoxic activity towards
BT 220, H116, Int 407, HL 251 and HLK 210 human cancer cells in vitro, tested by Apoptotic assay. 相似文献
4.
Rangarajulu Senthil Kumaran Heehoon Jung Hyung Joo Kim 《Engineering in Life Science》2011,11(3):264-271
The fungus Colletotrichum capsici was isolated from the diseased fruits of Chilli plant, Capsicum annuum. The isolated test fungus was identified by its morphological and molecular characteristic features. For the first time, the fungus was screened for the production of taxol on modified liquid medium. The presence of taxol was confirmed by the spectroscopic and chromatographic methods of analyses. The amount of taxol produced by this fungus was quantified by HPLC. The maximum amount of fungal taxol production was recorded as 687 μg/L. The production rate was 13 740‐fold higher than that, previously reported for the fungus Taxomyces andreanae. The extracted fungal taxol showed a strong cytotoxic activity in an in vitro culture of human cancer cells indicating that the increase in taxol concentration induces increased cell death. A PCR‐based screening for taxadiene synthase (ts), a unique gene in the formation of the taxane skeleton, confirmed the molecular blueprint for taxol biosynthesis. The results show that the fungus C. capsici 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. 相似文献
5.
Taxol production during the cultivation on a modified liquid and potato dextrose broth medium was indicated for the first time to occur in Phyllosticta spinarum, an endophytic fungus isolated from the needles of Cupressus sp. The presence of taxol in the fungal culture filtrate was confirmed by chromatographic and spectroscopic methods of analysis. The amount of taxol produced by this fungus was quantified by high performance liquid chromatography. The maximum amount of taxol production was obtained in this fungus when grown on M1D medium (235 μg/L) followed by PDB medium (125 μg/L). The results indicate that P. spinarum is an excellent candidate for taxol production . The production rate was 4.7 × 103‐fold higher than that found in the culture broth of an earlier reported fungus, Taxomyces andreanae. The fungal taxol extracted also showed a strong cytotoxic activity in the in vitro culture of human cancer cells tested in an apoptotic assay. 相似文献
6.
Rangarajulu Senthil Kumaran Johnpaul Muthumary Eun-Ki Kim Byung-Ki Hur 《Biotechnology and Bioprocess Engineering》2009,14(1):76-83
Taxol is a highly functionalized anticancer drug widely used in hospitals and clinics. The leaf spot fungus, Phyllosticta dioscoreae was isolated from diseased leaves of Hibiscus rosa-sinensis and screened for extracellular production of taxol in M1D (Modified liquid medium) and PDB (Potato dextrose broth) medium
for the first time. The fungus was identified by its morphological and conidial features in the culture growth. The presence
of taxol in the fungal culture filtrate was confirmed by different spectroscopic and chromatographic analyses. The amount
of taxol produced was quantified by HPLC. The maximum amount of taxol produced was found to be 298 μg/L in M1D medium. Production
rate was 5.96 × 103 times faster than that found in culture broth of earlier reported fungus, Taxomyces andreanae. The extracted fungal taxol also showed strong cytotoxic activity in vitro in the cultures of human cancer cells tested by apoptotic assay. The results indicate that P. dioscoreae is an excellent source of taxol production, which suggests that the fungus has potential to undergo genetic engineering in
order to improve its production level. 相似文献
7.
J Y Li R S Sidhu E J Ford D M Long W M Hess G A Strobel 《Journal of industrial microbiology & biotechnology》1998,20(5):259-264
A Periconia sp was isolated from Torreya grandifolia (a relative of yew that does not synthesize taxol) near Huangshan National Park in the People’s Republic of China. This fungus,
not previously known as a tree endophyte, was isolated from the inner bark of a small lower limb. When freshly isolated from
the tree and placed in a semi-synthetic medium, the fungus produced readily detectable quantities of the anticancer drug taxol.
Other taxol-producing endophytes were also isolated from this source. The production of taxol by Periconia sp was demonstrated unequivocally via spectroscopic and immunological methods. However, successive transfers of the fungus in
semi-synthetic medium resulted in gradual attenuation until low production occurred even though fungal growth was relatively
unaffected. Several compounds, known previously as activators of microbial metabolism, including serinol, p-hydroxybenzoic acid, and a mixture of phenolic acids, were capable of fully or partially restoring taxol production to otherwise
taxol-attenuated cultures. The compound with the most impressive ability to activate taxol production was benzoic acid at
0.01 mM. Benzoic acid was not a taxol precursor.
Received 19 December 1997/ Accepted in revised form 19 February 1998 相似文献
8.
Yan Chi Dong-Li Zhao Dong-Po Zhou 《World journal of microbiology & biotechnology》2008,24(11):2601-2605
The endophytic fungus Nodulisporium
sylviforme produces taxol, a diterpene alkaloid compound that has unique anti-neoplastic activity. In this study, suppression subtractive
hybridization was performed to identify genes in N. sylviforme that were involved in the up-regulation of taxol production. A total of 91 clones represented the differentially expressed
genes were selected based on reverse blotting results. These clones were then sequenced and analyzed using BLAST-X comparisons
that revealed that about 37.8% of the ESTs were homologous to 17 different proteins with known function, 20.7% were homologous
to nine hypothetical proteins and the remaining 41.5% had no identifiable homology; these may be novel genes. Two enzymatic
genes involved in taxol production were found. The data presented in this study provide the first primary overview of a set
of genes that are differentially expressed in N. sylviforme during taxol biosynthesis and form a solid foundation for the construction of high taxol-yielding genetically engineered
strains. 相似文献
9.
Paclitaxel production is enhanced in suspension‐cultured hazel (Corylus avellana L.) cells by using a combination of sugar,precursor, and elicitor 下载免费PDF全文
Sara‐Alsadat Rahpeyma Ahmad Moieni Mokhtar Jalali Javaran 《Engineering in Life Science》2015,15(2):234-242
Hazel (Corylus avellana L.) has recently been drawing attention as an alternative source of taxol. In the present study, the effects of sugar type, and different concentrations of phenylalanine (Phe) and vanadyl sulfate (V) on the production of taxol in C. avellana were investigated. A factorial experiment was used to optimize the concentrations of the precursor and elicitor. The cells were treated with Phe and V on the fourth day of culture and were harvested every 2 days until the 10th day. By increasing the Phe and V supply, taxol production increased during the culture period and the maximum level of 4.2 μg/g (dry weight) was obtained at day 10 by combining 3 μM of Phe and 0.05 and 0.1 mM of V in media supplemented with fructose (3%). The time course study on taxol production suggested that the appropriate time for using Phe is day 4 of culture, and day 8 for V. Overall, taxol production in C. avellana cell suspension culture was improved by the use of the combined strategy. 相似文献
10.
Zhao Kai Ping WenXiang Zhang LiNa Liu Jun Lin Yan Jin Tao Zhou DongPo 《中国科学C辑(英文版)》2008,51(3):222-231
To apply the fundamental principles of genome shuffling in breeding of taxol-producing fungi, Nodulisporium sylviform was used as starting strain in this work. The procedures of protoplast fusion and genome shuffling were studied. Three hereditarily
stable strains with high taxol production were obtained by four cycles of genome shuffling. The qualitative and quantitative
analysis of taxol produced was confirmed using thin-layer chromatography (TLC), high performance liquid chromatography (HPLC)
and LC-MS. A high taxol producing fungus, Nodulisporium sylviform F4-26, was obtained, which produced 516.37 μg/L taxol. This value is 64.41% higher than that of the starting strain NCEU-1
and 31.52%–44.72% higher than that of the parent strains. 相似文献
11.
Mingzi Wang Shaosong Liu Yaoyao Li Ren Xu Chunhua Lu Yuemao Shen 《Current microbiology》2010,61(4):254-260
Tubercularia sp. TF5 is an endophytic fungal strain isolated from the medicinal plant Taxus mairei. Previously, taxol has been detected in the fermentation products of this strain. However, it lost the capability of producing
taxol after long-term laboratory culture. Herein, we tried to reactivate the production of taxol by protoplast mutations and
genome shuffling. The protoplasts of Tub. sp. TF5 were prepared from its mycelia, and mutated by UV and NTG. The mutant strains regenerated from the mutated protoplasts
were selected and classified into four groups on the basis of their phenotypes, the profile of their metabolites analyzed
by TLC, MS, and bioassay data. Then, genome shuffling was subsequently carried out with eight mutant strains, with two representatives
from each protoplast mutant group, and genome shuffling mutant strains were obtained and screened using the same screening
procedure. Although taxol has not been detected in any mutant, two important mutants, M-741 and G-444 were selected for metabolites
isolation and determination due to their phenotypes, and differences in TLC analysis result from TF5 and other mutants. Three
new sesquiterpenoids, namely tuberculariols A–C (1–3), and a known dihydroisocoumarin (4) were obtained from M-741. Eighteen novel compounds were isolated from G-444, including five new sesquiterpenoids (5-9), two new dihydroisocoumarins (10, 11), one new tetralone (12), together with 10 known compounds (13–20, 1, and 2). The compounds isolated from the M-741 and G-444 were different in structure types and substitutions from those of TF5 (15, 21–29). The results showed, for the first time, that protoplast mutations and genome shuffling are efficient approaches to mining
natural products from endophytic fungi. Understanding the mechanisms of unlocking the biosynthesis of new metabolites will
facilitate the manipulation of the secondary metabolism in fungi. 相似文献
12.
JH Zhao YL Zhang LW Wang JY Wang CL Zhang 《World journal of microbiology & biotechnology》2012,28(5):2107-2112
An endophytic fungus was isolated from the root of the medicinal plant Moringa oleifera Lam. Based on analyzing the rDNA sequence, the fungus was identified as Nigrospora sp. This is the first report of the isolation of endophytic Nigrospora from M. oleifera. By bioassay-guided fractionation, four antifungal secondary metabolites were isolated from liquid cultures of the fungus
Nigrospora sp. LLGLM003, and their chemical structures were determined to be griseofulvin (1), dechlorogriseofulvin (2), 8-dihydroramulosin (3) and mellein (4) on the basis of spectroscopic analyses. Compound 2, 3 and 4 were isolated from Nigrospora sp. for the first time. In vitro antifungal assay showed that griseofulvin displayed clear inhibition of the growth of 8
plant pathogenic fungi. Dechlorogriseofulvin and mellein exhibited only weak antifungal activities, whereas 8-dihydroramulosin
displayed no antifungal activities. 相似文献
13.
AbstractAn experiment was conducted to study the effects of interaction between Meloidogyne incognita and Fusarium solani on plant length, fresh and dry weights, number of pods, chlorophyll, carotenoid, nitrogen and phosphorus contents and nitrate reductase activity in lentil plants. The results reveal a maximum damage occurring in all the plant growth, biochemical and nutrient parameters, in plants inoculated with M. incognita 10 days prior to F. solani (Mi?→?Fs). This was followed by simultaneous (Mi?+?Fs) inoculations, fungus inoculation 10 days prior to nematode (Fs?→?Mi), M. incognita alone and F. solani alone treatments. Nematode reproduction factor and root galling were highest in individual inoculation of M. incognita, while root rotting percentage was highest when nematode was inoculated 10 days prior to fungus followed by simultaneous inoculation with both nematode and fungus. 相似文献
14.
An endogenous filariform fungus has been separated from a tree named Taxus growing in the Aba region, Sichuan, China. The fungus is fermented in fluid medium for 3 weeks at 25℃, then the HPLC and MALDI-TOF analysis of the zymotic fluid show that the zymotic fluid contains taxol. So it is a fungus which can produce taxol. It is named Taxomyces sp. temporarily. 相似文献
15.
Bai Wan Deng Kai Hui Liu Wen Qiang Chen Xiao Wei Ding Xiu Chao Xie 《World journal of microbiology & biotechnology》2009,25(1):139-143
An endophytic fungus, Tax-3, was isolated from barks of Taxus chinensis grown in the Qinba mountains, China. The strain was classified into Fusarium solani based on the morphological characteristics and the molecular phylogenetics inferred from the nuclear ribosomal DNA ITS sequences
with the sequence similarity values of 100%. High performance liquid chromatography (HPLC) assay showed the F. solani, Tax-3, produced taxol with a higher yield of 163.35 μg/L in the reformative potato dextrose liquid medium (d), revealing
its potential applications for taxol production.
Bai Wan Deng and Kai Hui Liu contributed equally to this work. 相似文献
16.
Transcript profiling of jasmonate‐elicited Taxus cells reveals a β‐phenylalanine‐CoA ligase 下载免费PDF全文
Karla Ramírez‐Estrada Teresa Altabella Miriam Onrubia Elisabeth Moyano Cedric Notredame Lidia Osuna Robin Vanden Bossche Alain Goossens Rosa M. Cusido Javier Palazon 《Plant biotechnology journal》2016,14(1):85-96
Plant cell cultures constitute eco‐friendly biotechnological platforms for the production of plant secondary metabolites with pharmacological activities, as well as a suitable system for extending our knowledge of secondary metabolism. Despite the high added value of taxol and the importance of taxanes as anticancer compounds, several aspects of their biosynthesis remain unknown. In this work, a genomewide expression analysis of jasmonate‐elicited Taxus baccata cell cultures by complementary DNA‐amplified fragment length polymorphism (cDNA‐AFLP) indicated a correlation between an extensive elicitor‐induced genetic reprogramming and increased taxane production in the targeted cultures. Subsequent in silico analysis allowed us to identify 15 genes with a jasmonate‐induced differential expression as putative candidates for genes encoding enzymes involved in five unknown steps of taxane biosynthesis. Among them, the TB768 gene showed a strong homology, including a very similar predicted 3D structure, with other genes previously reported to encode acyl‐CoA ligases, thus suggesting a role in the formation of the taxol lateral chain. Functional analysis confirmed that the TB768 gene encodes an acyl‐CoA ligase that localizes to the cytoplasm and is able to convert β‐phenylalanine, as well as coumaric acid, into their respective derivative CoA esters. β‐phenylalanyl‐CoA is attached to baccatin III in one of the last steps of the taxol biosynthetic pathway. The identification of this gene will contribute to the establishment of sustainable taxol production systems through metabolic engineering or synthetic biology approaches. 相似文献
17.
18.
ZHAO Kai PING WenXiang ZHANG LiNa LIU Jun LIN Yan JIN Tao & ZHOU DongPo 《中国科学:生命科学英文版》2008,51(3):222-231
To apply the fundamental principles of genome shuffling in breeding of taxol-producing fungi, Nodulisporium sylviform was used as starting strain in this work. The procedures of protoplast fusion and genome shuffling were studied. Three hereditarily stable strains with high taxol production were obtained by four cycles of genome shuffling. The qualitative and quantitative analysis of taxol produced was confirmed using thin-layer chromatography (TLC), high performance liquid chromatography (HPLC) and LC-MS. A high taxol producing fungus, Nodulisporium sylviform F4-26, was obtained, which produced 516.37 μg/L taxol. This value is 64.41% higher than that of the starting strain NCEU-1 and 31.52%―44.72% higher than that of the parent strains. 相似文献
19.
一株产紫杉醇罗汉松内生真菌的分离和鉴定 总被引:4,自引:0,他引:4
[目的]紫杉醇是重要的抗癌药物,主要从罗汉松等植物中提取,为了保护罗汉松等种质资源,本文从罗汉松植株中分离产紫杉醇内生真菌,并对内生真菌所产紫杉醇的抗肿瘤活性进行了分析.[方法]采用组织块法自罗汉松的根、茎、叶等组织中分离内生真菌;通过四唑蓝(Methyl ThiazolylTetrazolium,MTT)比色法筛选有抗肿瘤活性的内生真菌菌株,通过薄层层析(Thin Layer Chro-matography,TLC)和高效液相色谱(High Performance Liquid Chromatography,HPLC)对内生真菌所产活性物质进行鉴定;采用抽提法抽提内生真菌所产紫杉醇,应用Vero细胞对抽提的紫杉醇的活性进行了分析.[结果]从罗汉松属(Podocrapus)植物中分离到155株内生真菌,其中28株内生真菌具有较高的抑癌活性.将其中一株菌株A2命名为EPTP-1,经形态学和分子分类学分析鉴定为烟曲霉(Aspergillus fumigatus).菌株EPTP-1中抽提的紫杉醇5.553μg/L~555.3 μg/L作用24h表现出明显的致细胞凋亡作用.菌株EPTP-1发酵5天时紫杉醇的产率为0.5578±0.0294 mg/L.[结论]从罗汉松中分离到了一株产紫杉醇内生真菌EPTP-1,可作为紫杉醇类药物工业化生产的候选菌株. 相似文献
20.
Ralf Oelmüller Irena Sherameti Swati Tripathi Ajit Varma 《Symbiosis (Philadelphia, Pa.)》2009,49(1):1-17
Piriformospora indica is a wide-host root-colonizing endophytic fungus which allows the plants to grow under extreme physical and nutrient stress.
The fungus can be cultivated on complex and minimal substrates. It belongs to the Sebacinales in Basidiomycota. P. indica has a vast geographical distribution and is reported from Asia, South America and Australia. The fungus is interesting for
basic research as well as biotechnological applications because: (i) it functions as a plant promoter and biofertilizer in
nutrient-deficient soils, (ii) as a bioprotector against biotic and abiotic stresses including root and leaf fungus pathogens
and insect invaders, (iii) as a bioregulator for plant growth development, early flowering, enhanced seed production, and
stimulation of active ingredients in medicinal plants (iv) as well as a bio-agent for the hardening of tissue-culture-raised
plants. Positive interaction are established for many plants of economic importance in arboriculture, agro-forestry, flori-horticulture
including Orchids, and those utilized for energy production and paper industry. P. indica also interacts with members of bryophyte, Aneura pinguis, pteridophyte, Pteris ensiormis, Gymnosperms (Pinus halepensis) and a large number of angiosperms (145 tested till date) including the model plant Arabidopsis thaliana and other members of the mustard family. Similar to arbuscular mycorrhizal fungi, P. indica stimulates nutrient uptake in the roots and solubilizes insoluble phosphatic and sulphur components in the soil. The interaction
of P. indica with the model plants Arabidopsis thaliana and barley (Hordeum vulgare L.) is used to understand the molecular basis of this beneficial plant/microbe interaction. We describe the current knowledge about the molecular basis of the interaction of plants with P. indica. An attempt is made to compare it with pathogenic and mycorrhizal plant/microbe interactions and also propose possible biotechnological
applications. 相似文献