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

Rangarajulu Senthil Kumaran,Johnpaul Muthumary,Byung-Ki Hur 한국식품과학회 2008 Food Science and Biotechnology Vol.17 No.6

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 ㎍/ℓ. The production rate was increased to 5.5×1,000 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.

Production of Taxol from Phyllosticta dioscoreae, a Leaf Spot Fungus Isolated from Hibiscus rosa-sinensis

Rangarajulu Senthil Kumaran,김은기,허병기,Johnpaul Muthumary 한국생물공학회 2009 Biotechnology and Bioprocess Engineering Vol.14 No.1

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 x 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.

Screening of species of the endophytic fungus<i>Phomopsis</i>for the production of the anticancer drug taxol

Kumaran, Rangarajulu Senthil,Hur, Byung‑,Ki Wiley (John WileySons) 2009 Biotechnology and Applied Biochemistry Vol.54 No.1

<P>Three different strains of the endophytic fungus Phomopsis were isolated from the healthy leaves of Taxus cuspidata (Japanese yew), Ginkgo biloba (ginkgo or maidenhair tree) and Larix leptolepis (Japanese larch) and screened for the production of taxol on a modified liquid medium for the first time. The presence of taxol was confirmed by spectroscopic and chromatographic analyses. The amount of taxol produced by this fungus was quantified by HPLC. The maximum amount of fungal taxol production was recorded as 418 microg/litre in the strain BKH 27. The yield was increased to 8360-fold that found for the fungus Taxomyces andreanae reported previously [Stierle, Strobel and Stierle (1993) Science 260, 214-216]. The fungal taxol extracted also showed a strong cytotoxicity towards the human cancer cells in an apoptosis assay. All the three isolates showed positive sign towards PCR for the conserved sequence of the taxadiene synthase gene. The results suggest that Phomopsis could be an excellent alternative source for taxol and may serve as a potential genetic-engineered species for the enhanced production of taxol.</P>

Quantitation of oxidative stress gene expression in MCF-7 human cell lines treated with water-dispersible CuO nanoparticles.

Kumaran, Rangarajulu Senthil,Choi, Yong-Keun,Kim, Hyung Joo,Kim, Kwang Jin Humana Press 2014 Applied biochemistry and biotechnology Vol.173 No.3

<P>The objective of this study was to assess the cytotoxicity of water-dispersible CuO nanoparticles by quantifying the reactive oxygen species (ROS)-related genes (glutathione S-transferase (GST) and catalase) using real-time polymerase chain reaction (RT-PCR). Monodisperse CuO nanoparticles of 14?nm in size were used. Cytotoxicity of CuO nanoparticles was evaluated under in vitro condition at different concentrations (10, 50, and 100?μg/ml) and incubation times (12, 24, and 48?h) with human cancer cell lines (breast cancer epithelial cells). The genetic level cytotoxic screening produced consistent results showing that GST and catalase ROS gene expression was maximized in 24?h incubation at 100?μg/ml concentration of CuO nanoparticles. However, the cytotoxicity of water-dispersible CuO nanoparticle was not significant compared with control experiments, demonstrating its high potential in the application of nanomedicines for a diagnostic and therapeutic tool.</P>

Cytotoxic Effects of ZnO Nanoparticles on the Expression of ROS-Responsive Genes in the Human Cell Lines

Kumaran, Rangarajulu Senthil,Choi, Yong-Keun,Singh, Vijay,Kim, Kwang Jin,Kim, Hyung Joo American Scientific Publishers 2016 Journal of nanoscience and nanotechnology Vol.16 No.1

<P>In the present investigation, engineered ZnO nanoparticles were tested for their induced oxidative stress in T47D tumor cell lines. The expressions of reactive oxygen species (ROS) related genes, glutathione S-transferase (GST) and catalase were quantified by real time-polymerase chain reaction (RT-PCR). In addition, qualitative analysis of GST was also performed at the cell level using molecular beacon (MB) technology. The tested nanoparticles were 20 nm in size, water-dispersible and treated on human breast tumor epithelial cell lines at 20, 40, 80 mu g/ml concentration with 14, 28, 48 h incubation times. Nanoparticles induced expressions of ROS responsive genes at molecular and cellular level, produces consistent results with respect to different dosage and incubation time. The experiment showed that the expression of both GST and catalase genes were maximized at 28 h with 80 mu g/ml concentration. However, the toxic effect of the monodisperse ZnO nanoparticles was not significant compared with control experiments, demonstrating its high potential in the applications of nanomedicines for a diagnostic and therapeutic tool.</P>

<i>In Vitro</i> Cytotoxic Evaluation of MgO Nanoparticles and Their Effect on the Expression of ROS Genes

Kumaran, Rangarajulu Senthil,Choi, Yong-Keun,Singh, Vijay,Song, Hak-Jin,Song, Kyung-Guen,Kim, Kwang Jin,Kim, Hyung Joo MDPI 2015 INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES Vol.16 No.4

<P>Water-dispersible MgO nanoparticles were tested to investigate their cytotoxic effects on oxidative stress gene expression. In this <I>in vitro</I> study, genes related to reactive oxygen species (ROS), glutathione <I>S</I>-transferase (GST) and catalase, were quantified using real-time polymerase chain reactions (molecular level) and molecular beacon technologies (cellular level). The monodispersed MgO nanoparticles, 20 nm in size, were used to treat human cancer cell lines (liver cancer epithelial cells) at different concentrations (25, 75 and 150 µg/mL) and incubation times (24, 48 and 72 h). Both the genetic and cellular cytotoxic screening methods produced consistent results, showing that GST and catalase ROS gene expression was maximized at 150 µg/mL nanoparticle treatment with 48 h incubation. However, the genotoxic effect of MgO nanoparticles was not significant compared with control experiments, which indicates its significant potential applications in nanomedicine as a diagnostic and therapeutic tool.</P>

Lipid body formation by Thraustochytrium aureum (ATCC 34304) in response to cell age

제은진,rangarajulu senthil kumaran,허병기 한국화학공학회 2008 Korean Journal of Chemical Engineering Vol.25 No.5

The heterotrophic marine protist, Thraustochytrium aureum produces substantial amounts of polyunsaturated fatty acids (PUFAs). In the present investigation, changes in the lipid and fatty acid profiles of T. aureum were studied according to the culture age. T. aureum was grown in artificial sea water medium for 10 days at 25 oC in shake culture condition. One to 10 day old cell samples were analyzed for cell biomass production, total lipid content, fatty acid profile and lipid body formation. In all the samples tested, total lipid production was found to be directly proportional to the dry cell weight of T. aureum. In the early phase of cell growth, cell biomass production, lipid content and glucose consumption were found to be higher. Thin layer chromatographic analysis (TLC) of lipids showed the presence of triacylglycerol (TAG; 169 mg/g, 90%), phospholipids (PL; 83mg/g, 66%) and sterol (ST; 6 mg/g, 5%), which were recorded at maximum levels in the early growth phase of the cells. The composition of PUFAs and saturated fatty acids (SFAs) of the cell biomass and lipid class components (TAG and PL) was identified by gas chromatographic analysis (GC). In the early phase of cell growth, production of PUFAs in the total fatty acids was found to have attained maximum levels (61.3%) in which docosahexaenoic acid alone showed higher content of occurrence (99.0 mg/g in total lipid; 65.2 mg/g in TAG and 41.0 mg/g in PL). In the middle phase of cell growth, palmitic acid production was found to be higher (36.7 mg/g in total lipid; 31.3mg/g in TAG and 12.6mg/g in PL). Transmission electron microscopic studies of the cells showed the presence of a membrane around the lipid bodies in the early phase of cell growth. TAG and PL were actively involved in the formation of lipid bodies in the cells of T. aureum. Large-sized lipid bodies accumulated in 3 day old cells which were then fragmented into smaller bodies in the late growth phase.

Electrochemical Control of Methane Emission from Lake Sediment Using Microbial Fuel Cells

Hyeon Jin Jeon,최용근,rangarajulu senthil kumaran,김성현,송경근,홍석원,김미아,김형주 대한화학회 2012 Bulletin of the Korean Chemical Society Vol.33 No.7

Lipid Body Formation and Fatty Acid Production of the Protist, Thraustochytrium aureum ATCC 34304

Eun-Jin Jeh,Rangarajulu Senthil Kumaran,Byung-Hee Chun,Byung-Ki Hur 한국생물공학회 2007 한국생물공학회 학술대회 Vol.2007 No.4

LED light stress induced biomass and fatty acid production in microalgal biosystem, <i>Acutodesmus obliquus</i>

Choi, Yong-Keun,Kumaran, Rangarajulu Senthil,Jeon, Hyeon Jin,Song, Hak-Jin,Yang, Yung-Hun,Lee, Sang Hyun,Song, Kyung-Guen,Kim, Kwang Jin,Singh, Vijay,Kim, Hyung Joo Elsevier 2015 Spectrochimica acta. Part A, Molecular and biomole Vol.145 No.-

<P><B>Abstract</B></P> <P>Microbial algal system can serve as a potential source for the production of much high value bioproducts and biofuels. The quality and intensity of light are the key elements to optimize the production of algal biomass and fatty acid contents. This study presents the effect of differential LED flashing light conditions on the growth of microalgae, <I>Acutodesmus obliquus</I>. The induced light stress was optimized for its biomass and fatty acid content. The microalgae are exposed to various frequency of intermittent LED flashing light (blue and red lights) at three different phases in the 18day cell growth (log, lag and stationary phase). The frequency of light flashing rate was adjusted to 120, 10, 5, 3.75, and 1times per min. The effect of light stress on growth and fatty acids composition of <I>A. obliquus</I> induced an increase in algae growth and fatty acid production. Different optimal timing for light stress was subjected to elucidate the effect of light stress on algae growth and fatty acid production. The results showed an increase in the algae growth (1.2mg/L of chl a content) under light stress condition at FT10 (flashing time, 10times per min) from the initial day (log phase) compared with the control experiment (0.4mg/L of chl a content). However, the total fatty acids (71mg/g) and volumetric FAME production (9.4ml/l) level was found to be significant under FT5 (flashing time, 5times per min), adopting flashing light from day 10 (stationary phase). TEM studies also revealed the deposition of lipid to be largest in the 18day old cells under flashing light (FT5) condition, representing maximum accumulation of lipids bodies (up to 770nm diameter in particle size) occupying approximately 42% of the total area of the cell.</P> <P><B>Highlights</B></P> <P> <UL> <LI> LED light stress on algal biomass and fatty acid production were studied. </LI> <LI> Flashing lights exposed on log, lag, and stationary phases. </LI> <LI> FT10 showed an increase in the algae growth. </LI> <LI> Total fatty acids and volumetric FAME production were higher under FT5. </LI> <LI> Deposition of lipid was found maximum in the 18day old cells (FT5). </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>