Figure 1.
Morphological features of endophytic fungus Fusarium oxysporum.
Colony shape: (a) Mycelium, (b) Chlamydospores (c) Macroconidia, microconidia and chlamydospores formed on PDA.
Figure 2.
(a) TLC of crude fungal vinblastine from culture filtrates along with standard Vinblastine 1: Vinblastine standard, 2: Crude sample, 3: Vincristine standard, Detection: Ceric ammonium sulphate reagent. (b) TLC of partially purified fungal vinblastine from culture filtrates along with standard vinblastine on silica gel using chloroform∶methanol (8∶2) solvent system. A: Standard vinblastine B: Partially purified vinblastine C: Partially purified vincristine, Detection: Ceric ammonium sulphate reagent. (c) TLC of fungal vinblastine purified from culture filtrates along with standard vinblastine on silica gel using chloroform∶methanol (8∶2) solvent system. 1: Purified fungal vinblastine 2: Standard vinblastine, Detection: Ceric ammonium sulphate reagent. (d) TLC of fungal vincristine purified from culture filtrates along with standard vincristine on silica gel using chloroform∶methanol (8∶2) solvent system. 1: Standard vincristine, 2: Purified fungal vincristine, Detection: Ceric ammonium sulphate reagent.
Figure 3.
(a) HPLC profile of pure fungal vinblastine with retention time of 36.6 min. (b) HPLC profile of pure fungal vincristine with retention time of 34.9 min. (c) UV absorption spectrum of standard vinblastine and fungal vinblastine. (d) UV absorption spectrum of standard vincristine and fungal vincristine.
Figure 4.
(a) Molecular mass determination of the fungal vinblastine by ESI-MS. (b) Molecular mass determination of the fungal vincristine by ESI-MS.
Figure 5.
(a) MS-MS spectrum of the purified fungal vinblastine. (b) MS-MS spectrum of the purified fungal vincristine.
Figure 6.
(a) 400 MHz 1H NMR spectra of fungal vinblastine (A) and standard vinblastine sulphate (B). The signals marked with ‘s’ is coming from the residual solvent (CHCl3). The signals marked with * and x are due to water (from the solvent) and contamination form n- hexane, respectively. The broad signal marked ú′ at ∼6 ppm in A is an unidentified impurity. (b) 500 MHz 1H NMR spectra of fungal vincristine (A) and standard vincristine sulphate (B). The signals marked with ‘s’ are coming from the residual solvent (Methanol-d4). The signals marked with * and x are due to contamination from trace amounts of n-hexane and ethanol, respectively.
Table 1.
Proton NMR (1H NMR) chemical shifts of standard vinblastine and fungal vinblastine in CDCl3.
Table 2.
Proton NMR (1HNMR) chemical shifts of standard vincristine sulphate and fungal vincristine in Methanol-d4.