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Amylase (E.C.3.2.1.1) are the enzyme that can catalyse the hydrolysis of starch into simple monomers. Amylase enzymes are widely needed for use in various industrial fields such as the textile, food, paper and other industries. Yeast can produce enzymes more effectively and safer than other organisms. Amylolytic yeast can be isolated from flower substrates because it contains sugar for yeast nutritional needs. This study aims to isolate, select and characterize amylolytic yeast on the substrate of Paphiopedilum sp. from Bedugul Botanical Garden, Bali. Yeast isolation was carried out with direct and washing method. Screening was carried out on YPSA medium with diffusion agar method. The result of isolation obtained 19 isolates of yeast with characteristics of 73.9% white-mucoid, 21.05% cream-mucoid and 5.26% light flesh-mucoid. Screening results showed that 10 isolates were able to produce an amylase enzyme with code isolates P1, P4, P6, P10, P11, P12, P13, P14, P15 and P19. The potential yeast isolates in yielding amylase with P12 isolate codes had amylolytic index 0,45 mm.