For efficient lincomycin production from Streptomyces lincolnensis L1245, various vegetable oils, natural nitrogen sources, and surfactants were investigated at the pilot-scale level in the flask. Olive oil as the sole carbon source was the most suitable one for producing lincomycin. When 20 g/lof olive oil was used, the lincomycin concentration and lipase activity reached 1.01 g/land 182 U/ml, respectively, after 5 days of culture. Among the various unsaturated fatty acids, when linolenic acid was used, the cell growth and lincomycin production were markedly decreased. On the other hand, when 0.2 g/l of oleic acid was added to the culture broth, the maximum lincomycin concentration was 1.0 g/l, which was about 1.7-fold higher than that obtained without the addition of oleic acid. Among the various natural nitrogen sources, pharmamedia or soybean meal was the most suitable nitrogen source. In particular, in the case of a mixture of 10 g/l of pharmamedia and soybean meal, 1.5 g/l of lincomycin concentration and 220 U/ml of lipase activity were obtained. When Span 180 was used as the surfactant, lincomycin production, lipase activity, and oil consumption increased. The correlation between the consumption rates of oil and lincomycin production in a culture using olive oil as the sole carbon source was also investigated. The lincomycin production depended on the consumption rate of olive oil. Using these results, fed-batch cultures for comparing the use of olive oil and starch as a conventional carbon source were carried out in a 5-1 fermentor. When olive oil was used as the sole carbon source, 34 g/l of olive oil was consumed after 7 days of culture. The maximum lincomycin concentration was 3.0 g/l, which was about 2.0-fold higher than that of starch medium after 7 days of culture. The product yield was 0.09 gig of consumed carbon source, which was about 3.0-fold higher than that of starch medium after 7 days of culture.