A lipid producing isolate (LPI 65) was identified by a molecular genetics technique as Pichia occidentalis MH879824.1 was investigated for its ability to produce biodiesel. The isolate was screened for its ability to accumulate lipids within the cells by culturing on nitrogen-limited medium (NLM) and using Sudan black B staining technique. Estimation of lipid content % was performed. The effect of different glucose concentrations in NLM on dry biomass and lipid weight were conducted and results showed that the best concentration of glucose was 70 g/L. The possibility of using different wastes such as sugar cane molasses, waste food oil and shortening, whey permeate, and waste motor oil as carbon sources for lipid production were determined, and the results showed that the best waste for lipid production was waste motor oil. Optimization of fermentation conditions such as effect of NLM composition, NLM pH, incubation temperature, and incubation period were also investigated and results of these tests showed that the best lipid accumulation when NLM5, at pH 5.5 and incubated at 28oC for 120 h. Fatty acids profile analysis achieved that the lipids extracted from the isolate mainly contained the principal fatty acids (triacylglycerol’s, TAGs) which are similar to that of vegetable oils mainly oleic, linoleic and palmitic acid.

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