Antibacterial metabolites from Bipolaris specifera, an endophytic fungus from the endemic medicinal plant, Zingiber nimmonii (J. Graham) Dalzell

Eleven fungal endophytes were isolated from the plant parts of Z. nimmonii (J. Graham) Dalzell, an endemic species of the Western Ghats, India, a biodiversity hotspot area. The endophytic isolates were characterized by the sequencing of the Internal Transcribed Spacer (ITS) regions and designated as strains by depositing ITS sequences in the Gen Bank sequence database. All the strains were cultured in Potato Dextrose broth (PDB, 500 mL) contained in Erlenmeyer flasks to obtain the secondary metabolites. The culture filtrate was extracted with ethyl-acetate (EA) three times and concentrated by flash evaporation to obtain EA crude dry extract. The strains were evaluated for the antibacterial potentials against six pathogenic bacterial strains viz., Bacillus subtilis (MTCC 121), Staphylococcus aureus (MTCC 7443) Pseudomonas aeruginosa (MTCC 7093), Escherichia coli (MTCC 729), Enterobacter aerogenes (MTCC 111) and Klebsiella pneumoniae (MTCC 661). Nine endophytic fungal extracts except Alternaria consortiale and Hypocrea lixi showed inhibitory activities against at least two of the six test bacterial strains. Bipolaris specifera (KM114290) exhibited the highest inhibition zones ranging from 15.1 ± 0.3 to 26.7 ± 1.1 mm (diameter), against all six test bacteria in the agar disk diffusion assay, and with Minimum Inhibitory Concentrations (MIC's) of 0.04-0.14 mg/mL, followed by Aspergillus terreus. B. specifera extract was therefore selected and characterized for the identification of antibacterial compounds by chromatographic techniques. Seven antibacterial compounds viz., (1) Bicyclo[3.2.0]heptan-2-one, 6-hydroxy-5-methyl-6-vinyl; (2) Adipic acid divinyl ester; (3) 1,4-Naphthoquinone, 6-acetyl-2,5-dihydroxy; (4) Decanedioic acid, 3,7-dimethyl ester; (5) (Z)-4-Hexenoic acid 2-acetyl-2-methyl-ethyl ester and (6) Butanoic acid 2-acetyl-3-methyl-methyl ester and (7) Caffeic acid, were identified through liquid and gas chromatography. These compounds are mainly volatile esters of fatty acids, phenolics and adipic acid found rare in nature. This study envisages the possible drug discovery using endophytes from traditional and endemic medicinal species.

Keywords: Antibacterial; Bipolaris specifera; Endophytic fungi; GC–MS; Western Ghats; Zingiber.