Isolation and Partial Characterization of Bioactive Components of Endophytic Fungi Penicillium singorense, Isolated from Two Indian Medicinal Plants: Calotropis procera and Catharanthus roseus

Debjani Roy Chowdhury¹, Swapan Kumar Chattopadhyay² and SubhashKanti Roy^1,

¹PG Department of Biotecnology and Biochemistry, Oriental Institute of Science and Technology (Vidyasagar University), Dewandighi, Katwa Road, Burdwan-02, West Bengal, India

²PG Department of Botany, Hooghly Mohsin College, Chinsurah, Hooghly, West Bengal, India

Cite this paper:
Debjani Roy Chowdhury, Swapan Kumar Chattopadhyay and SubhashKanti Roy. Isolation and Partial Characterization of Bioactive Components of Endophytic Fungi Penicillium singorense, Isolated from Two Indian Medicinal Plants: Calotropis procera and Catharanthus roseus. American Journal of Microbiological Research. 2022; 10(3):84-93. doi: 10.12691/ajmr-10-3-2

Abstract

Fungal endophytes are recently gaining momentum for alternative source of plant secondary metabolites having potent bioactivity. In this study, the objective of which was to isolate and partial characterize the bioactive chemical component present in the cell free extract of endophytic fungi Penicillium singorense isolated from Calotropis procera and Catharanthus roseus. The fungus (pure culture of Penicillium singorense DRC1, Accssession No. MG322179) was cultured in PDB (Potato Dextrose Broth) media. The culture filtrates were obtained by filtering through Whatman 42 filter paper placed on a Buchner funnel fitted to a vacuum pump. Presence of taxol in the fungal extract was detected following the standard protocol as described by Shrinivasan and Muthumary. The detection of taxol was carried out following the standard protocol of UV spectroscopy, Thin Layer Chromatography and High Performance Liquid Chrmatography. The antibiotic like compound was purified following the standard procedure involving transfer of the compound repeatedly from ethyl acetate phase to aqueous phase, then precipitation. The antibiotic like compound was extracted in ethyl acetate which was then allowed to evaporate to dryness in a rotary vacuum evaporator at 35.5°C, residue was weighed. The partially purified compound was subjected to IR, NMR and GCMASS spectroscopy for partial characterization. The cell free extract of P. singorense isolated from C. roseus and C. procera indicated the presence of highly valued anticancerous diterpenoid compound ‘Taxol’. Proton NMR study indicated that the compounds may be constituted with an aromatic benzene ring trisubstituents at positions 1, 3 and 4; the remainder (unintegrated) look like aliphatic (alkyl chain) protons. The FTIR spectroscopic study revealed that the molecule can have an aromatic skeleton with different substituents. The GCMS study revealed the presence of 9 major compounds (most commonly occurring of those are Methyl stearate (C₁₉H₃₈O₂₎, 1-Nonadecene (C₁₉H₃₈) and 3- Eicosene (C₂₀H₄₀)), maximum of which are of aliphatic in nature.

Keywords:
Endophytic fungi Penicillium singorense Accssession No. MG322179 Taxol bioactive compound Methyl stearate (C₁₉H₃₈O₂) 1-Nonadecene (C₁₉H₃₈) 3- Eicosene (C₂₀H₄₀)

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