Yarrowia lipolytica,one of the unconventional yeasts,grows well and produces erythritol with a high formation rate using glycerol as a sole carbon source. High osmotic pressure generated by NaCl and glycerol benefitted to enhance erythritol production,but high osmotic pressure would inhibit yeast cell growth,extend the period of fermentation,and drop the productivity down. In this research,glycine and proline were used as the osmoprotectants to investigate how the hyperosmotic resistance of Y. lipolytica was improved. The results indicated that Y. lipolytica could import extracellular amino acid and accumulate as a compatible solute in cytoplasm in order to resist the damages of the high osmotic pressure. Consequently,the erythritol synthesis and the biomass were enhanced remarkably. In 7 L stirred fermenter test with the 4. 17 ± 0. 17 osmol / kg of initial osmotic pressure,when 30 mg / L glycine and 40 mg / L proline were added at the beginning of the fermentation,the fermentation period was reduced from 108 h to 90 h,and the final concentration,productivity and yield of erythritol were 93. 6 ± 4. 2 g / L,1. 04 ± 0. 05 g /( L· h) and 0. 49 ± 0. 03 g / g,which had increased 4. 12%,25. 3% and 4. 26%,respectively.