A cold-tolerant fungal strain Thelebolus microsporus was investigated for the first time for its pigment and fatty acid production. High-performance liquid chromatography analysis confirmed the presence of carotenoid pigment. Gas chromatography observations showed the presence of major fatty acids: myristic acid (14:0), palmitic acid (16:0), stearic acid (18:0), heptadecanoic acid (17:0), linolenic acid (18:3) and linoleic acid (18:2). Of these, linolenic acid, a polyunsaturated fatty acid, was present in substantial quantity, suggesting that it may have a role in adapting the fungus to low Antarctic temperatures by modulating membrane fluidity. The commercial application of linolenic acid is as a food supplement for humans suffering from eczema, cardiovascular disease and diabetic neuropathy. Another fatty acid, linoleic acid, a precursor of 1-octen-3-ol, is the principal aromatic compound in most fungi and has also been documented in this strain. Screening of the fungal culture for extracellular enzyme activity for amylase, protease, lipase, chitinase and cellulase was carried out. The isolate showed maximum α-amylase activity at 20°C, suggesting effective applications as a detergent additive, in textile processing and in the food industry.