Journal of Microbiology

Byung-Ki Hur

2 Articles

Isolation and Identification of an Anticancer Drug, Taxol from Phyllosticta tabernaemontanae, a Leaf Spot Fungus of an Angiosperm, Wrightia tinctoria: Rangarajulu Senthil Kumaran , Johnpaul Muthumary , Byung-Ki Hur; J. Microbiol. 2009;47(1):40-49. Published online February 20, 2009; DOI: https://doi.org/10.1007/s12275-008-0127-x

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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 (M1D) 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 M1D medium (461 ug/L) followed by PDB medium (150 ug/L). The production rate was increased to 9.2x103 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.

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Study on Persistent Infection of Japanese Encephalitis Virus Beijing-1 Strain in Serum-free Sf9 Cell Cultures: Hun Kim , Su Jeen Lee , Jin Yong Park , Yong Wook Park , Hyun Sung Kim , Heui-Yun Kang , Byung-Ki Hur , Yeon-Woo Ryu , Sang In Han , Jong Su Kim; J. Microbiol. 2004;42(1):25-31.; DOI: https://doi.org/2005 [pii]

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Abstract PDF: Sf9 cells have obvious advantages for the conventional production technology of vaccine. They are useful tools for high concentration and large-scale cultures. Sf9 cells were grown to maximal concentration, 8×10^6 cells/ml in a 500ml spinner flask, with a doubling time at the exponentially growing phase of 24.5 hours, using serum-free media. To explore the ability of Sf9 cells to be infected by the Japanese encephalitis (JE) virus Beijing-1 strain, Sf9 cells were infected with the virus. By 4-5 days post-infection, 10-15% of the Sf9 cells showed cytopathic effect (CPE), from granularity to the formation of syncytia and multinucleated giant cells continuously observed over a period of 35 days. Positive fluorescent reactions were detected in 30-40% of cells infected with the JE virus Beijing-1 strain, and the uninfected Sf9 cells were completely negative. Virus particles, propagated in Sf9 and Vero cells, were concentrated by sedimentation on 40% trehalose cushions by ultracentrifugation, and showed identical patterns of viral morphogenesis. Complete virus particles, 40 to 50 nm in diameter, were observed, and JE virus envelope (E) proteins, at 53 kDa, were found in the western blot analysis to the anti-JE virus E protein monoclonal antibody and reacted as a magenta band in the same position to the glycoprotein staining. To evaluate whether the infectious virus was produced in Sf9 cells inoculated with the JE virus Beijing-1 stain, Sf9 cells were inoculated with the virus, and sample harvested every 5 days. The titers of the JE virus Beijing-1 strain rose from 1.0×10^5 to 1.5×10^6 pfu/ml. The infected Sf9 cells could be subcultured in serum-free medium, with no change in the plaque sizes formed by the JE virus Beijing-1 strain in the plaque assay. It is suggested that the ability of the JE virus Beijing-1 strain to infect Sf9 cells in serum-free media will provide a useful insect cell system, where the JE virus replication, cytopathogenicity and vaccine immunogen can be studied.

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