b-Glucosidases have great potential to be used in various biotechnological processes, such as those used by the food and feed industries, biomass hydrolysis for bioethanol production, and enhancing the flavor of wine, tea, and fruit juice. This work describes b-glucosidase production by Penicillium miczynskii. The effects of pH, time, and condition of cultivation, temperature, different carbon sources, and concentration of the selected agro-industrial waste on extracellular b-glucosidase production were studied in submerged fermentation. Additionally, the enzyme produced was biochemically characterized. The best conditions for P. miczynskii b-glucosidase production were 3% pineapple peel as carbon source, under stationary conditions, for 9 days, at pH 5.5 and temperature of 20°C, which yielded 2.82 U/mL. The enzyme showed best activity at pH 4.5-5.0, and 65°C. It was stable at 55°C and 60°C, with a half-life of 50 min and 40 min, respectively. The bglucosidase was slightly activated in the presence of bmercaptoethanol, Ca 2 + , and Co 2 + , and strongly inhibited by sodium dodecylsulfate (SDS), Zn 2 + , and Hg 2 + . Also, sodium citrate, NH 4 + , and Ba 2 + were able to inhibit this enzyme, especially at 10 mM concentration. The enzyme presented high glucose tolerance, with a K i of 760 mM. This study describes a novel bglucosidase that presents favorable industrial properties, such as low inhibition rate by glucose, and production on pineapple peel, an inexpensive and abundant agro-industrial waste.