Methicillin-resistant Staphylococcus epidermidis (MRSE) and methicillin-resistant Staphylococcus haemolyticus (MRSHa) are important coagulase-negative staphylococci. They are often isolated from bacteremia in humans mainly due to their ability to form biofilm on the surfaces of medical devices. Papain is a complex mixture of proteolytic enzymes and peroxidases extracted from the latex of Carica papaya and it is recognized by accelerating the healing process of wounds. This study aimed to evaluate the ability of the MRSE and MRSHa isolates to produce biofilms. Besides this, the ability of papain to inhibit the formation of biofilms or to disrupt the ones already formed by those bacteria was analyzed. Thirty MRSHa and 30 MRSE were isolated from bacteremia and used in this study. It was observed that papain has ability to reduce biofilms formed by MRSE (p < 0.06) and by MRSHa (p = 0.0005). In addition, papain was able to disrupt mature biofilms made by MRSE (p = 0.014). No antibacterial activity of papain was observed for any isolates of MRSE and MRSHa tested. Papain has been demonstrated as a potential product for reducing biofilm. Uniterms:Staphylococcus epidermidis/methicillin-resistant. Staphylococcus haemolyticus/methicillinresistant. Biofilm/production. Papain/reducing biofilm.Staphylococcus epidermidis resistente à meticilina (MRSE) e Staphylococcus haemolyticus resistente à meticilina (MRSHa) são importantes estafilococos coagulase negativa. São frequentemente isolados em bacteremia humana, principalmente devido à capacidade de formar biofilmes nas superfícies de dispositivos médicos introduzidos no organismo. A papaína é mistura complexa de enzimas proteolíticas e peroxidases extraídas do látex de Carica papaya, reconhecida por acelerar os processos de cura de feridas. Este estudo teve como objetivo avaliar a capacidade dos MRSE e MRSHa em produzir biofilmes e analisar a capacidade da papaína em inibir a formação de biofilme ou desintegrar biofilmes já formados por essas bactérias. Observou-se que a papaína tem capacidade de reduzir a formação de biofilme por MRSE (p < 0,06) e MRSHa (p = 0,0005). Além disso, a papaína foi capaz de desintegrar biofilme maduro formado por MRSE (p = 0,014). Nenhuma atividade antibacteriana da papaína foi observada para qualquer das duas espécies de bactérias testadas. A papaína mostrou-se produto potencial para reduzir biofilme.Unitermos: Papaína. Staphylococcus epidermidis/resistente à meticilina. Staphylococcus haemolyticus/ resistente à meticilina. Biofilme/produção. Papaína/redução de biofilme. INTRODUCTIONCoagulase-negative staphylococci, especially Staphylococcus epidermidis followed by Staphylococcus haemolyticus, have been frequently reported as significant etiological agents that cause nosocomial infections (Keim et al., 2011, Kristóf et al., 2011Rosado, Romanelli, Camargos, 2011). The most important property of these staphylococci is their capability to form biofilm on the surfaces of foreign bodies introduced into the organism (Donlan, 2011;Sander et al., 2012). Th...