Eucommia leaf contains large amounts of natural active products. In extracting the substances, the most important is the removal of the cuticle layer on the leaves and the cell wall in the leaves of Eucommia ulmoides. But the removal of the cuticle layer is a technical difficulty now. Cutinase (EC3.1.1.74) is a multifunctional enzyme with a common alpha/beta fold structure belonging to hydroplane that can make a substantial degradation of horny fatty acids. So this study isolated bacteria capable of producing cutinase from the lesion of Eucommia leaves and identified the bacteria. The identification using PCR-RFLP method confirmed that the strain belongs to Rhodotorula mucilaginosa. The fermentation conditions of the strain-producing cutinase were optimized in this study. The finding of cutinase-producing R. mucilaginosa is significant because the yeast is more secure than plant pathogens, being suitable for mass production.