The entophytic microorganisms will be the big interest in the enzymes production of biotechnological interest. However, little things in literature about entophytic yeasts invertase producers exist. The aim of this work was isolate endophytic yeasts invertase producers from the intracellular spaces and vacuoles. The yeasts was isolated from the stem (YS) and root (YR) of Saccharum officinarum and the culture had been carried out in Erlenmeyers (250 mL) with 10g of sugar cane in 50 mL of distilled water, in orbital shaker (180 rpm) at 30 for 48h. The enzyme was obtained of the supernatant after extraction with a phosphate buffer solution 0,2 M pH 5,0 and centrifuged at 10000 xg for 20 minutes. The analytical determinations had been carried for invertase activity and sacarose consumption. The biomass production was determined through absorbance (600nm). The maximum activities of invertase had been gotten at 36 hours of culture for yeasts, 419 and 379 U/mL for YS and YR, respectively. In the consumption of carbon source, the yeasts had reduced about 30% of the reducing sugar to the end 24 hours. This can be due to affinity to the substrate (high concentration sacarose) of invertase produced by the yeasts. The results demonstrate the potential of these yeasts for invertase production with great affinity for high concentrations of sacarose, what it can make possible process of production of inverted sugar in industrial scale.