Exopolysaccharide analysis of biofilm‐forming Candida albicans

Abstract

The major objective of the study was to analyse exopolysaccharide produced by a biofilm forming-clinical strain of Candida albicans. The biofilm-forming ability of C. albicans recovered from infected intrauterine devices (IUDs) was evaluated using XTT (2,3-bis[2-methoxy-4-nitro-5-sulfophenyl]-2H-tetrazolium-5-carboxanilide) reduction assay. The morphological characteristics of the biofilm were assessed using scanning electron microscopy (SEM), atomic force microscopy (AFM) and confocal laser scanning microscopy (CLSM). Biochemical characterization of the exopolysaccharide was carried out by gel permeation chromatography, gas chromatography (GC), Fourier transform infrared (FTIR) spectroscopy and nuclear magnetic resonance (NMR) spectroscopy. Microscopic studies of C. albicans biofilm revealed complex, heterogeneous three-dimensional structure, in which yeast cells and hyphal elements were entrenched within exopolysaccharides matrix. Chromatographic analysis data indicated C. albicans exopolysaccharide (c. 300 kDa) to be made up of four major sugar units. The FTIR spectrum revealed specific absorbance of O-H, C-H, O=C=O, C=O, C-N and C-C ring stretching. (1) H and (13) C NMR data showed the presence of β (1→6) and β (1→3) linkages in the exopolysaccharide chain that were assigned to α-D-glucose and β-D-glucose, α-D-mannose, α-L-rhamnose and N-acetyl glucosamine (β-D-GlcNAc), respectively. Study suggested the production of a water soluble c. 300 kDa exopolysaccharide by C. albicans made up of glucose, mannose, rhamnose and N-acetyl glucosamine subunits. The study could assist in the development of novel therapeutics aimed at disrupting C. albicans biofilms that will translate into improved clearance of Candida-related infections.