Browsing by Author "Weerasinghe, W. R. H."

Now showing 1 - 2 of 2

Bioactive properties and metabolite profile of an endolichenic fungus, Hypoxylon lividipigmentum
(Faculty of Science, University of Kelaniya, Sri Lanka., 2021) Weerasinghe, W. R. H.; Shevkar, C. D.; Silva, R. S. De; Attanayake, R. N.; Weerakoon, G.; Kate, A. S.; Kalia, K.; Paranagama, P. A.
Endolichenic fungi (ELF) serve as a novel source of secondary metabolites. Hypoxylon lividipigmentum is an ELF isolated from the lichen Opegrapha medusulina, collected from mangrove plant Xylocarpus granatum from Negombo lagoon, Sri Lanka. The fungus was identified to the species level using morphological and DNA barcoding techniques. Ethyl acetate extract of the fungus was subjected to in vitro assays to determine antioxidant, anti-inflammatory, tyrosinase inhibitory and antibacterial potency. Liquid Chromatography-Mass Spectrometry (LCMS) dereplication was conducted on the crude extract in order to detect the secondary metabolites present. The extract reported a IC50 value of 18.34±1.37 µg/ml on par with the positive control BHT, in DPPH radical scavenging assay. It also exhibited moderate anti-inflammatory activity with an IC50 value of 81.08±1.05 μg/ml. Tyrosinase inhibitory activity was fairly comparable with an IC50 value of 121.20±2.55 μg/ml. Agar well diffusion assay was conducted to determine antibacterial activity against aerobic bacterial species Escherichia coli, Bacillus subtilis, Staphylococcus aureus and the anaerobic bacterial species Streptococcus mutans. Suppression of growth was shown only against B. subtilis. Five major mass peaks were observed during the study of LCMS profile of the extract. After a thorough dereplication process, two masses could be presumed to be from novel scaffolds. Since none of the mass peaks could be dereplicated within the species or genus level, it could be speculated that the chemical profile of Hypoxylon lividipigmentum was previously poorly explored in literature thus making it an interesting organism to study further for novel metabolites.
Exploring bioactive compounds in the endolichenic fungus, Xylaria feejeensis, inhabiting the lichen, Graphis librata, collected from Negombo lagoon, Sri Lanka
(Institute of Chemistry Ceylon Adamantane House, Rajagiriya, Sri Lanka., 2020) Gunawardhana, M. H. A. Y.; Wickramarachchi, P. A. S. R.; Weerasinghe, W. R. H.; Paranagama, P. A.
The accelerating pharmaceutical problem of bacteria growing resistant to existing antibiotics forces the scientific community to search for new antibacterial compounds for antibiotic drug development. Literature reveals that Sri Lankan mangrove inhabiting endolichenic fungal (ELF) population is rich in many such bioactive compounds. Previously isolated and cultured ELF, Xylaria feejeensis; from the lichen Graphis librata inhabiting in the mangrove plant Rhizophora mucronata was cultured on 60 large potato dextrose agar medium containing petri dishes and incubated for 14 days at room temperature. After extracting secondary metabolites to ethyl acetate (EA) the resulted crude extract was tested for its antibacterial and antioxidant activity. EA crude extract showed negligible antioxidant activity in DPPH radical scavenging assay hence this assay was not proceeded further. The activity of crude extract (5 mg/ml) against Escherichia coli (ATCC25922), Staphylococcus aureus (ATCC25923) and Bacillus subtilis (ATCC6051) was tested using agar well diffusion antibacterial assay. An inhibition zone diameter of 1.9 cm, 2.2 cm, and 2.2 cm against three bacterial strains with Azithromycin positive control (5 mg/ml) 2.2 cm, 2.2 cm and 2.3 cm was observed respectively. By partitioning of EA crude extract resulted two antibacterial active hexane and chloroform fractions and one antibacterial inactive 60% methanol in water fraction. Hexane fraction showed the highest antibacterial activity with inhibition diameter zone of 2.6 cm against E.coli with 2.6 cm diameter of inhibition zone for positive control. Further purification of hexane fraction was performed by normal phase column chromatography and 4 fractions were resulted. The highest polar fraction showed 1.4, 2.1 and 1.7 cm diameter inhibition zones of growth of E.coli, S. aureus and B. subtilis comparable with 2.2 cm Azithromycin. The other 3 fractions showed no antibacterial activity against these bacterial strains. Results suggest that further purification of active fraction and structure elucidation might result in new antibiotic lead compounds.