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<p>Vinblastine and vincristine are clinically important anti-cancer alkaloids traditionally sourced from <i>Catharanthus roseus</i>, though plant-based extraction is often limited by low yield and high production costs. This study aimed to identify and characterize an endophytic fungal alternative capable of produce these compounds under laboratory conditions. An endophytic fungus isolated from <i>Catharanthus roseus</i> (L.) G.Don showed potential and tested positive for the tryptophan decarboxylase (TDC) gene. It was identified as <i>Epichloe festucae</i>, and its ability to produce vinblastine and vincristine was confirmed by electrospray ionization mass spectrometry (ESI–MS) with molecular mass of vinblastine 811&#xa0;m/z and vincristine 824&#xa0;m/z respectively and quantification performed via high-performance liquid chromatography (HPLC). Culturing of endophytic fungus in vinca medium resulted in the production of 61.5&#xa0;µg/L of vinblastine and 52.8&#xa0;µg/L of vincristine. The alkaloids were purified using silica gel column chromatography and structurally characterized by Fourier-transform infrared spectroscopy (FTIR), ultraviolet (UV) spectroscopy and nuclear magnetic resonance (NMR). The cytotoxicity of the partially purified compounds was evaluated against HeLa and MCF-7 cancer cell lines using the MTT assay. Vinblastine exhibited IC₅₀ values of 25.5&#xa0;µg/mL (HeLa) and 24.73&#xa0;µg/mL (MCF-7), while vincristine showed IC₅₀ values of 35.9&#xa0;µg/mL and 38.8&#xa0;µg/mL, respectively. Apoptosis-inducing activity was further validated in HeLa cells via Annexin V/PI staining, Acridine orange/Ethidium bromide dual staining, and cell cycle analysis using PI/Rnase staining with fluorescence-activated cell sorting (FACS). These findings demonstrated that <i>Epichloe festucae</i> is a promising microbial platform for the sustainable production of vinblastine and vincristine, offering a potential alternative to plant-derived sources for commercial and pharmaceutical applications.</p>

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Endophytic fungus Epichloe festucae from Catharanthus roseus as a novel source of vinblastine and vincristine with apoptosis-inducing activity

  • 期刊论文

  • 发表日期：

    2025年10月14日

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  • E. Swathi,
  • M. S. Vineetha,
  • Sunil S. More

摘要

Vinblastine and vincristine are clinically important anti-cancer alkaloids traditionally sourced from Catharanthus roseus, though plant-based extraction is often limited by low yield and high production costs. This study aimed to identify and characterize an endophytic fungal alternative capable of produce these compounds under laboratory conditions. An endophytic fungus isolated from Catharanthus roseus (L.) G.Don showed potential and tested positive for the tryptophan decarboxylase (TDC) gene. It was identified as Epichloe festucae, and its ability to produce vinblastine and vincristine was confirmed by electrospray ionization mass spectrometry (ESI–MS) with molecular mass of vinblastine 811 m/z and vincristine 824 m/z respectively and quantification performed via high-performance liquid chromatography (HPLC). Culturing of endophytic fungus in vinca medium resulted in the production of 61.5 µg/L of vinblastine and 52.8 µg/L of vincristine. The alkaloids were purified using silica gel column chromatography and structurally characterized by Fourier-transform infrared spectroscopy (FTIR), ultraviolet (UV) spectroscopy and nuclear magnetic resonance (NMR). The cytotoxicity of the partially purified compounds was evaluated against HeLa and MCF-7 cancer cell lines using the MTT assay. Vinblastine exhibited IC₅₀ values of 25.5 µg/mL (HeLa) and 24.73 µg/mL (MCF-7), while vincristine showed IC₅₀ values of 35.9 µg/mL and 38.8 µg/mL, respectively. Apoptosis-inducing activity was further validated in HeLa cells via Annexin V/PI staining, Acridine orange/Ethidium bromide dual staining, and cell cycle analysis using PI/Rnase staining with fluorescence-activated cell sorting (FACS). These findings demonstrated that Epichloe festucae is a promising microbial platform for the sustainable production of vinblastine and vincristine, offering a potential alternative to plant-derived sources for commercial and pharmaceutical applications.