The isolated fungus, designated as LB‐54, was found to be most suitable for the production of GLA due mainly to its high content of lipid, 20.73 % of dry cell weight and GLA of 14.68 % of total fatty acids. The strain LB‐54 was identified asMucorsp on the basis of morphological characteristics. The influence of different carbon and nitrogen sources, incubation time and growth temperature was investigated. Glucose and yeast extract were found to be the best carbon and nitrogen sources, respectively, in terms of GLA yield. The highest production of GLA (74 mg/1) was obtained from the LB‐54 strain after 5 days of incubation at 28°C in basal medium containing glucose (20 g/1) and yeast extract (10 g/1) following glucose addition (7 % w/v) and incubation for an additional 3 days at 12°C.