Fatty acids are main important for building prostaglandins, which are necessary for several body processes, including dilation of blood vessels, cholesterol metabolization and other critical bio-chemical functions and physiological reaction. The total lipids determination were detected in supernatant of each hydrolysis cells and cultures. Important variation were recorded between the extracellular and intracellular total lipids for all yeast strains. All strains were grown at the same condition but with Yeast peptone Glucose (YPG) medium and 3 % glucose. For selecting the highest total lipid producing strains, results showed that the strains GT160.34B and STX23-5B were the highest for lipid of extracellular (7.5 and 10.2 g/l), respectively. While the intracellular lipid of the tested strains GT160-34B, GM3 and C321RE221R were the higher where reached to (14 g/l). The tested yeast strains showed considerable range in the iodine values thatindicatedvarying amounts of unsaturated fatty acids. The two hybrids and protoplasts strains; F (GT160-34B× GM3), H₁(GT160-34B× STX23-5B) and H₂(GT160-34B×STX23-5B) were higher producers of the extracellular, intracellular fatty acid and total lipid concentration, respectively. . The conclusion of these data, it's provided that the improved strains by (protoplast fusion and hypradzition), F(GT160-34B×GM3), H₁(GT160-34B×STX23-5B) and H₂(GT160-34B×STX23-5B) were better strains contained fatty acids comparison with the organal strains.