Nanoflowers are recently developed flower-shaped nanoparticles consisting of several layers of petals to improve surface reaction and stability. Among them organic-inorganic hybrid nanoflowers have been spotlighted, since they can overcome enzyme-related limitations, including low stability, high production cost, substrate/product inhibition and difficult recovery. In this study, triple enzyme-inorganic hybrid nanoflowers (TrpE@ihNFs) were synthesized using a mixture of three enzymes (alpha-amylase, lipase and protease) and Cu2+ ions. TrpE@ihNFs were characterized by their morphology and chemical point of view by using different techniques including SEM, FTIR, EDX, and XRD. Afterwards we compared the enzyme activity and stability of TrpE@ihNFs with the free enzymes, including lipase, amylase and protease at the different pH and temperatures spectrophotometrically. Our data reveal that enzyme activities and stability of TrpE@ihNFs were significantly higher compared to the each free enzyme. In conclusion, we showed TrpE@ihNFs which can be used for the treatment of wastewater, biosensors, biocatalysts, and bio-related devices in the future. (C) 2020, The Society for Biotechnology, Japan. All rights reserved.